Application of covalent organic framework material in preparation of blood perfusion adsorbent

Abstract

The invention relates to application of a covalent organic framework material in preparation of a blood perfusion adsorbent, and the covalent organic framework material not only can adsorb toxins in blood with high adsorption capacity, but also has the advantages of excellent adsorption selectivity, low hemolysis rate, protein adhesion resistance and the like.

Description

Technical Field

[0001] The present invention relates to the field of medical devices, in particular to an adsorbent in a blood perfusion device, and specifically to the use of a covalent organic framework material in preparing a blood perfusion adsorbent. Background Art

[0002] Blood purification technology is a treatment method that uses an extracorporeal blood circulation device to directly remove endogenous and exogenous toxins from the patient's blood, and then returns the purified blood to the patient's body. It can replace the patient's own detoxification system to remove toxins and achieve artificial intervention on blood components. As blood purification technology is applicable to more and more diseases, blood purification therapy is known as the "third therapy" after drug therapy and surgical therapy.

[0003] At present, the blood purification treatment methods that have been developed mainly include hemodialysis, hemofiltration, hemoperfusion, blood exchange, etc. Hemodialysis uses membrane materials to separate blood and dialysate, uses the concentration difference on both sides of the membrane as the driving force, and removes toxins through pore size screening. Blood purification technology based on the principle of membrane separation is effective for removing small molecule toxins in the blood, but it is not ideal for removing protein-bound toxins or large molecule toxins. Hemoperfusion is a blood purification technology based on the principle of adsorption separation, which can efficiently remove protein-bound toxins and large molecule toxins in the blood.

[0004] The core of hemoperfusion technology is the performance of the adsorbent material used in the hemoperfusion device. An ideal adsorbent material needs to have a high adsorption rate, a large adsorption capacity, good blood compatibility and high mechanical strength. At present, the most commonly used adsorbent materials are activated carbon, polysaccharides and resin materials. Activated carbon materials have the advantages of large specific surface area, low cost and easy availability, but their blood compatibility is poor and the adsorption selectivity is low. Polysaccharide materials have good blood compatibility and can also improve selectivity by modifying functional groups, but their adsorption capacity is low and their mechanical strength is poor. Resin materials are some of the most widely used adsorbent materials at present. They have high mechanical strength and are easy to process, but their adsorption capacity and selectivity still need to be improved. For example, in 2016, Howell et al. developed dextran-coated activated carbon for the adsorption of vitamin B12 and bilirubin. The biocompatibility of dextran-coated activated carbon was improved, but the dextran coating blocked the pores of the activated carbon, and its bilirubin adsorption capacity was only 8.7 mg / g. In 2017, Wu et al. prepared chitosan-CNT-lysine composite microspheres with good cell compatibility and blood compatibility, but its ability to remove bilirubin in plasma was 0.363 mg / mL. In 2017, Chen et al. prepared polystyrene resin (NKA-9) by suspension polymerization, and its adsorption capacity for bilirubin was 4.3 mg / mL.

[0005] The above-mentioned currently commonly used hemoperfusion adsorbent materials still have deficiencies in adsorption capacity, adsorption selectivity and blood compatibility to varying degrees. Therefore, it is very important to develop new hemoperfusion adsorbent materials with excellent adsorption performance and good blood compatibility.

[0006] Covalent organic frameworks (COFs) are a type of crystalline polymer materials formed by long-range orderly connections of organic monomers through covalent bonds. The pore size of COFs is uniform, so they are also called "organic zeolites". Compared with inorganic porous materials, COFs have the following characteristics: 1) Structural diversity. By selecting building blocks with different geometric shapes or changing the connection method of building blocks, frameworks with different topological structures can be obtained; 2) Open and controllable pores. The pore size, shape and chemical environment can be regulated through structural design and functionalization; 3) The organic molecules that make up COFs are connected by strong covalent bonds (such as BO, C=N, CN, C=C, etc.), which makes COFs not easily attacked by chemical reagents or easily dissociated by high temperatures, and thus have high chemical stability and thermal stability. The above characteristics make COFs show great application potential in the fields of separation, sensing, catalysis, energy storage, etc. Summary of the invention

[0007] In view of the problems in the prior art, the present invention provides a new use of a covalent organic framework material to solve the problems of low adsorption capacity, poor blood compatibility, poor selectivity, etc. of the existing blood perfusion adsorbent materials.

[0008] One aspect of the present invention provides a use of a covalent organic framework material in preparing a blood perfusion adsorbent.

[0009] Preferably, the covalent organic framework material is formed by reacting a first monomer comprising an amino group and a second monomer comprising an aldehyde group.

[0010] More preferably, the first monomer is 1,3,5-tris(4-aminophenyl)benzene, tetrakis-(4-aminophenyl)ethylene, 1,3,6,8-tetrakis(4-aminophenyl)pyrene or 2,4,6-tris(4-aminophenyl)-1,3,5-triazine.

[0011] More preferably, the second monomer is phthalaldehyde, 2,5-di-hydroxyterephthalaldehyde or 2,5-bis(prop-2-yn-1-yloxy)terephthalaldehyde.

[0012] More preferably, the covalent organic framework material is one or more of the following: The covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and terephthalaldehyde. The X-ray powder diffraction spectrum of the formed covalent organic framework material is as follows: Figure 1 As shown; The covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and 2,5-di-hydroxyterephthalaldehyde. The X-ray powder diffraction spectrum of the formed covalent organic framework material is as follows: Figure 8 As shown; The covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and terephthalaldehyde. The X-ray powder diffraction spectrum of the formed covalent organic framework material is as follows: Fig.15 As shown; The covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)benzene and 2,5-bis(prop-2-yn-1-yloxy)terephthalaldehyde. The X-ray powder diffraction spectrum of the formed covalent organic framework material is as follows: Fig. 22 As shown; The covalent organic framework material is formed by the reaction of 1,3,6,8-tetrakis(4-aminophenyl)pyrene and 2,5-di-hydroxyterephthalaldehyde. The X-ray powder diffraction spectrum of the formed covalent organic framework material is as follows: Fig.29 As shown; The covalent organic framework material is formed by the reaction of tetrakis-(4-aminophenyl)ethylene and terephthalaldehyde. The X-ray powder diffraction spectrum of the formed covalent organic framework material is as follows: Fig.36 shown.

[0013] Preferably, the target adsorbent of the hemoperfusion adsorbent is protein-bound toxins and / or macromolecular toxins in the patient's blood.

[0014] More preferably, the target adsorbed substance is bilirubin and / or bile acid in the patient's blood.

[0015] Preferably, the pore size of the covalent organic framework material is larger than the diameter of the target adsorbent of the hemoperfusion adsorbent.

[0016] Preferably, the covalent organic framework material is used in combination with a second adsorption material, and the second adsorption material is one or more selected from the group consisting of activated carbon, polysaccharides, resins, and immunosorbents.

[0017] More preferably, the immunoadsorbent is an antigen-antibody binding adsorbent, a complement binding adsorbent, an Fc binding adsorbent, an electrostatic binding adsorbent or a hydrophobic binding adsorbent.

[0018] The inventors have found that when the covalent organic framework material is used as a blood perfusion adsorbent, it not only has a high adsorption capacity for the target adsorbate, for example, the adsorption amount of bilirubin can be as high as 155.6 mg / g, but also has excellent adsorption selectivity, low hemolysis rate and anti-protein adhesion. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. Together with the following specific embodiments, they are used to explain the present invention, but do not constitute a limitation of the present invention.

[0020] Figure 1 is the X-ray powder diffraction pattern of COF-A; Figure 2 is the infrared spectrum of COF-A; Figure 3 N of COF-A 2 Adsorption and desorption curves; Figure 4 This is the kinetic curve of bilirubin adsorption of COF-A; Figure 5 This is the bile acid adsorption kinetic curve of COF-A; Figure 6 is the albumin adsorption graph of COF-A; Figure 7 is the hemolysis rate spectrum of COF-A; Figure 8 is the X-ray powder diffraction pattern of COF-B; Fig. 9 is the infrared spectrum of COF-B; Fig.10 N of COF-B 2 Adsorption and desorption curves; Fig.11 This is the kinetic curve of bilirubin adsorption of COF-B; Fig.12 This is the bile acid adsorption kinetic curve of COF-B; Fig.13 is the albumin adsorption diagram of COF-B; Fig.14 is the hemolysis rate spectrum of COF-B; Fig.15 is the X-ray powder diffraction pattern of COF-C; Fig.16 is the infrared spectrum of COF-C; Fig.17 N of COF-C 2 Adsorption and desorption curves; Fig.18 This is the kinetic curve of bilirubin adsorption of COF-C; Fig.19 This is the bile acid adsorption kinetic curve of COF-C; Fig. 20 is the albumin adsorption diagram of COF-C; Fig.21 is the hemolysis rate spectrum of COF-C; Fig. 22 is the X-ray powder diffraction pattern of COF-D; Fig.23 is the infrared spectrum of COF-D; Fig.24 N of COF-D 2 Adsorption and desorption curves; Fig.25 This is the kinetic curve of bilirubin adsorption of COF-D; Fig.26 This is the bile acid adsorption kinetic curve of COF-D; Fig. 27 is the albumin adsorption graph of COF-D; Fig.28 is the hemolysis rate spectrum of COF-D; Fig.29 is the X-ray powder diffraction pattern of COF-E; Fig.30 is the infrared spectrum of COF-E; Fig.31 N of COF-E 2 Adsorption and desorption curves; Fig.32 This is the kinetic curve of bilirubin adsorption of COF-E; Fig.33 This is the bile acid adsorption kinetic curve of COF-E; Fig.34 is the albumin adsorption graph of COF-E; Fig.35 is the hemolysis rate spectrum of COF-E; Fig.36 is the X-ray powder diffraction pattern of COF-F; Fig.37 is the infrared spectrum of COF-F; Fig.38 N of COF-F 2 Adsorption and desorption curves; Fig.39 This is the kinetic curve of bilirubin adsorption of COF-F; Fig.40 This is the bile acid adsorption kinetic curve of COF-F; Fig.41 is the albumin adsorption graph of COF-F; Fig.42 This is the hemolysis rate spectrum of COF-F. DETAILED DESCRIPTION

[0021] The specific implementation of the present invention is described in detail below in conjunction with the accompanying drawings. It should be understood that the specific implementation described here is only used to illustrate and explain the present invention, and is not used to limit the present invention.

[0022] The terms used in the present invention have the meanings generally used in the art. Some technical terms used in the present invention are explained and defined below.

[0023] Covalent organic framework materials The "covalent organic framework" material in the present invention, also known as COFs, is a type of crystalline polymer material formed by long-range orderly connection of organic monomers through covalent bonds. The pore size of the covalent organic framework material is uniform, so it is also called "organic zeolite". The covalent organic framework material has the characteristics of regular and adjustable pores, large specific surface area, high porosity, good stability, and easy functionalization. The covalent organic framework material used in the present invention can be synthesized by methods known in the art, such as Xiong Chen et al. Towards covalent organic frameworks with predesignable and aligned open docking sites. Chem. Commun., 2014, 50, 6161-6163, DOI: 10.1039 / c4cc01825g; Laura Ascherl et al. Molecular docking sites designed for the generation of highly crystalline covalent organic frameworks. Nature Chemistry, 2016, DOI: 10.1038 / NCHEM.2444. The above patents or documents are fully introduced into the present invention.

[0024] Hemoperfusion Hemoperfusion is a blood purification technology that introduces the patient's blood into a perfusion device filled with adsorbents to remove exogenous or endogenous toxins, drugs or metabolic wastes in the blood that cannot be removed by dialysis through adsorption. Unlike hemodialysis based on the principle of membrane separation, hemoperfusion can efficiently remove protein-bound toxins and large molecular toxins in the blood. Among them, protein-bound toxins refer to toxins that exist mostly in a bound form in plasma and accumulate due to abnormal metabolism under pathological conditions. Since the free level of protein-bound toxins in the blood is low, the toxins bound to proteins cannot pass through the dialysis membrane and are therefore difficult to remove through conventional hemodialysis technology. For example, uremic protein-bound toxins (PBUTs) accumulate in patients with end-stage renal disease, among which indoxyl sulfate and p-cresol sulfate have been shown to be associated with increased all-cause mortality and cardiovascular disease in patients with chronic kidney disease (CKD); for patients with abnormal liver function, bilirubin levels are an independent risk factor for death in patients with acute-on-chronic liver failure, and accumulated protein-bound toxins play an important role in the secondary development of severe complications such as renal failure, hepatic encephalopathy, and circulatory disorders; bile acid levels are elevated in patients with acute-on-chronic liver failure (ACLF). Large molecular toxins such as parathyroid hormone, serum creatinine, and urea nitrogen.

[0025] Hemoperfusion adsorbent Hemoperfusion adsorbent is a core component in hemoperfusion technology. It is installed in the hemoperfusion device. When the patient's blood is introduced into the hemoperfusion device outside the body, it adsorbs exogenous or endogenous toxins, drugs or metabolic wastes. When selecting materials for preparing hemoperfusion adsorbents, it is usually necessary to consider the adsorption capacity, selectivity, hemolysis rate, anti-protein adhesion, etc. of the material for the target adsorbent in the patient's blood. In the present invention, the inventors surprisingly found that covalent organic framework materials can be used as an excellent hemoperfusion adsorbent, which not only has a high adsorption capacity, but also has excellent selectivity, hemolysis rate, anti-protein adhesion, etc. In addition, the commonly used hemoperfusion adsorbents include activated carbon, resin, polysaccharide, immunosorbent, etc. Among them, activated carbon can be petroleum charcoal, resin charcoal, and mother-and-child capsule charcoal. Petroleum charcoal: prepared by forming, oxidizing, and activating petroleum colloid and asphalt. Resin charcoal: charcoal formed by oxidation, carbonization, and activation of synthetic resin. Mother-and-child capsule charcoal: natural fruit shells are prepared into 300-mesh carbon powder, and microcapsules are formed using microcapsule technology. The shape of activated carbon can be irregular granular, spherical, columnar, fibrous, powdery, etc. The resin can be an ion exchange resin with polar exchange groups or an adsorption resin without exchangeable groups. The immunosorbent can be an antigen-antibody binding adsorbent, a complement binding adsorbent, an Fc binding adsorbent, an electrostatic binding adsorbent or a hydrophobic binding adsorbent. Among them, the antigen-antibody binding adsorbent is to bind the antigen to a fixed carrier to adsorb the corresponding antibody or immune complex in the blood, such as anti-acetylcholine receptor antibody. Complement binding adsorbents, for example, use C1q to bind to the Fc segment of the immune complex to adsorb immune complexes, such as DNA-anti-DNA complexes. Fc binding adsorbents, such as Staphylococcus aureus protein A adsorbents, can bind to IgG1, IgG2, IgG4, and Fc fragments in immunoglobulins, and can also bind to IgG3, IgM, IgA, and Fab fragments in a non-immune reaction manner. Electrostatic binding adsorbents refer to adsorbents that bind to adsorbed substances by electrostatic action to remove adsorbed substances. For example, dextran sulfate contains polyanions, so it can effectively remove anti-ds-DNA antibodies, antiphospholipid antibodies, low-density lipoproteins, antithrombin III, C3a, etc. Hydrophobic binding adsorbents, such as cross-linking phenylalanine or tryptophan to the hydroxyl groups of polyvinyl alcohol gels with microporous structures, can relatively specifically remove pathogenic factors in a hydrophobic manner.

[0026] The present invention is described in detail below.

[0027] The present invention provides a use of a covalent organic framework material in the preparation of a hemoperfusion adsorbent. The inventors of the present invention have found that the covalent organic framework material can adsorb target adsorbates (such as protein-bound toxins and / or macromolecular toxins) in the patient's blood with a high adsorption capacity, such as the adsorption of bilirubin can be as high as 155.6 mg / g. At the same time, the covalent organic framework material has excellent selectivity, low hemolysis rate, and anti-protein adhesion. Therefore, the covalent organic framework material can be used as a hemoperfusion adsorbent with superior performance.

[0028] The covalent organic framework material is composed of organic monomers connected by covalent bonds. Those skilled in the art can select different organic monomers and adjust their pore size, pore environment, pore shape, porosity, etc. by methods known in the art. Preferably, the covalent organic framework material is obtained by reacting a first monomer containing an amino group and a second monomer containing an aldehyde group. The covalent organic framework material synthesized from the first monomer and the second monomer has better mechanical properties and is more suitable for use as a blood perfusion adsorbent.

[0029] More preferably, the first monomer is 1,3,5-tris(4-aminophenyl)benzene, tetrakis-(4-aminophenyl)ethylene, 1,3,6,8-tetrakis(4-aminophenyl)pyrene or 2,4,6-tris(4-aminophenyl)-1,3,5-triazine, and the second monomer is phthalaldehyde, 2,5-di-hydroxyterephthalaldehyde or 2,5-bis(prop-2-yn-1-yloxy)terephthalaldehyde. More preferably, the covalent organic framework material is one or more of the following: the covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and terephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is as follows: Figure 1 As shown; the covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and 2,5-di-hydroxyterephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is as shown Figure 8 As shown; the covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and terephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is as shown Fig.15 As shown; the covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)benzene and 2,5-bis(prop-2-yn-1-yloxy)terephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is as shown Fig. 22 As shown; the covalent organic framework material is formed by the reaction of 1,3,6,8-tetrakis(4-aminophenyl)pyrene and 2,5-di-hydroxyterephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is as shown Fig.29As shown; the covalent organic framework material is formed by the reaction of tetrakis-(4-aminophenyl)ethylene and terephthalaldehyde, and the X-ray powder diffraction pattern of the covalent organic framework material formed is as shown Fig.36 shown.

[0030] Preferably, the pore size of the covalent organic framework material is larger than the diameter of the target adsorbent of the hemoperfusion adsorbent, so that the target adsorbent can be adsorbed more effectively.

[0031] When the covalent organic framework material is used to prepare the blood perfusion adsorbent, the covalent organic framework material can be in any shape such as powder, spherical or irregular granular, fibrous, etc., and the above shapes will not affect the adsorption effect of the covalent organic framework material.

[0032] In addition, the covalent organic framework material of the present invention can be used in combination with a second adsorbent material, and the second adsorbent material can be selected from materials known in the prior art for preparing blood perfusion adsorbents, for example, the second adsorbent material is selected from one or more of the group consisting of activated carbon, polysaccharides, resins, and immunosorbents. Among them, the activated carbon can be petroleum charcoal, resin charcoal, or mother capsule charcoal, and the immunosorbent can be an antigen-antibody binding adsorbent, a complement binding adsorbent, an Fc binding adsorbent, an electrostatic binding adsorbent, or a hydrophobic binding adsorbent. For example, when the covalent organic framework material is used in combination with the second adsorbent material, the constructed blood perfusion adsorbent is a multilayer structure, in which the covalent organic framework material layer and the second adsorbent material layer are alternately arranged. Or the covalent organic framework material is doped with the second adsorbent material and fixed by chemical bonds / covalent bonds before use. Without affecting the technical effect of the present invention, those skilled in the art can combine the covalent organic framework material with the second adsorbent material by a commonly used combination method.

[0033] The covalent organic framework materials used in the embodiments of the present invention can be synthesized using methods known in the art, and their structures and properties can be characterized using an infrared spectrometer, an X-ray powder diffractometer, a gas adsorption instrument, a scanning electron microscope, an ultraviolet spectrophotometer, and an automatic coagulation instrument.

[0034] 1. Structure and basic properties of covalent organic framework materials The structure of the covalent organic framework material was characterized using an infrared spectrometer, an X-ray powder diffractometer, and a scanning electron microscope. The specific instruments and parameters are as follows: Infrared spectrometer: Bruker ALPHA, wavelength range 400 cm -1 ~ 4000 cm -1, Bruker Company, USA. X-ray powder diffractometer: Model Bruker Foucus D8, Bruker Company, USA; wherein, the powder sample scanning temperature is 298 K, the pressure is 40 kV, the current is 50 mA, and the X-ray radiation source is Cu-Kα.

[0035] The pores of the covalent organic framework material were characterized using a gas adsorption instrument. The instrument and parameters are as follows: Gas adsorption instrument: Model Quantachrome (ASiQMVH002-5), Quantachrome Company, USA; the adsorption amount of nitrogen by the prepared material was tested under standard atmospheric pressure (101 kPa), and the purity of the gas used in the test was 99.999%.

[0036] 2. Toxin removal performance test of covalent organic framework materials In the present invention, bilirubin and bile acid were taken as examples to test the toxin removal performance of the covalent organic framework material.

[0037] Bilirubin removal performance test A certain amount of covalent organic framework material was weighed and immersed in a vial containing 10 mL of a certain concentration of bilirubin / albumin composite solution, wherein bilirubin was purchased from Beijing Solebao and albumin (BSA) was purchased from Anage. The solvent of the composite solution was 0.1M phosphate buffer solution. After sealing, it was placed in a constant temperature water bath shaker at 37 o C and oscillate at a certain frequency for a certain time. Centrifuge and take the supernatant, and use a fully automatic blood biochemical analyzer (Mindray BS-350s) to determine the total bilirubin concentration. The test process was carried out under light-proof conditions. Among them, the dosage of the covalent organic framework material was 5-10 mg, the bilirubin concentration range was 100-200 mg / L, the albumin concentration range was 30-50 g / L, and the oscillation frequency range was 150-200 rpm.

[0038] Bile acid removal performance test Weigh a certain amount of covalent organic framework material and immerse it in a vial containing 10 mL of a certain concentration of bile acid / albumin complex solution. After sealing, place it in a constant temperature water bath shaker at 37 o C and oscillate at a certain frequency for a certain time. Centrifuge and take the supernatant, and use a fully automatic blood biochemical analyzer (Mindray BS-350s) to measure the total bile acid concentration. The amount of covalent organic framework material is 5-10 mg, the bile acid concentration range is 100-200 mg / L, the albumin concentration range is 30-50 g / L, and the oscillation frequency range is 150-200 rpm.

[0039] 3. Adsorption selectivity test A certain amount of covalent organic framework material was mixed with plasma from patients with liver disease (the plasma came from the waste plasma from plasma exchange treatment of patients in the hospital) or blood from normal people, and then stirred in a constant temperature water bath shaker at 37 o C and oscillate at a certain frequency for a certain time, centrifuge and take the supernatant, and use a fully automatic blood biochemical analyzer (Mindray BS-350s) to detect various biochemical indicators in plasma. Among them, the amount of covalent organic framework material is 2-10 mg, the amount of plasma is 2-10 mL, the oscillation frequency range is 150-200 rpm, and the oscillation time range is 1-3 h.

[0040] 4. Blood compatibility test of covalent organic framework materials Hemolysis test Test group: A certain amount of covalent organic framework material was mixed with 2 mL of 6% sheep red blood cell solution (purchased from Beijing Solebao) and incubated at 37 o C, stand for 1 h, centrifuge and take the supernatant, and use an ultraviolet spectrophotometer (model UV-2600, Shimadzu Corporation, Japan) to measure its absorbance at 541 nm. The amount of the covalent organic framework material used is 2-10 mg.

[0041] Positive control: The covalent organic framework material in the test group was replaced with an equal mass of water and tested with the same steps as the above test group.

[0042] Negative control: The covalent organic framework material in the test group was replaced with an equal mass of isotonic solution (0.1 M phosphate buffer solution) and tested with the same steps as the above test group.

[0043] The hemolysis rate was calculated as follows: (absorbance of the test group - absorbance of the negative control) / (absorbance of the positive control - absorbance of the negative control).

[0044] Anti-protein adhesion test Weigh a certain amount of covalent organic framework material and immerse it in a vial containing 10 mL of a certain concentration of albumin solution (solvent is 0.1M phosphate buffer solution). After sealing, place it in a constant temperature water bath shaker at 37 o C and oscillate at a certain frequency for a certain time. Centrifuge and take the supernatant, and use an ultraviolet spectrophotometer (model UV-2600, Shimadzu Corporation, Japan) to measure its absorbance at 280nm. The amount of covalent organic framework material is 2-10 mg, the albumin concentration range is 30-50 g / L, the oscillation frequency range is 150-200 rpm, and the oscillation time range is 3 h.

[0045] The following describes the implementation of the present invention through specific embodiments, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific implementations, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. Example Example 1

[0046] 30 mg of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and 17 mg of terephthalaldehyde were dissolved in a mixed solution of 3 mL of anhydrous ethanol and 3 mL of o-dichlorobenzene, dispersed by ultrasonication, and then 0.25 mL of 3M acetic acid was added and shaken. After three freeze-pump-thaw cycles, the container was sealed and then heated at 120 o C for 3 days in an undisturbed environment. As the reaction proceeds, a precipitate is gradually generated. The mother liquor is removed by centrifugation, and the precipitate is collected and washed 3 times with tetrahydrofuran (THF), replaced 3 times with anhydrous ethanol, and then heated at 120 o C and vacuum dried for 24 h to obtain the covalent organic framework material COF-A.

[0047] Figure 1 is the X-ray powder diffraction pattern of COF-A, Figure 1 Curve 1 in the figure represents the simulation curve of COF-A. Those skilled in the art can obtain the simulation curve of X-ray powder diffraction of the covalent organic framework material involved in each embodiment of the present invention through a commonly used database, such as the database CSDS of the Cambridge Crystal Data Center CCDC. Curve 2 represents the measurement curve of the synthesized COF-A. The characteristic peak in curve 2 coincides with curve 1, indicating that the above-mentioned covalent organic framework material has been successfully synthesized.

[0048] Figure 2 is the infrared spectrum of COF-A, where 1640 cm -1 The appearance of the characteristic peak of imine bond indicated that the monomers were covalently bound.

[0049] Figure 3 N of COF-A 2 The adsorption and desorption curves show that the adsorption isotherm curve presents type IV, indicating that COF-F has a mesoporous structure.

[0050] Figure 4 The kinetic curve of bilirubin adsorption by COF-A is shown in Figure 1, where the dosage of the covalent organic framework material is 10 mg, the bilirubin concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, the adsorption amount of bilirubin by COF-A (q t) reached nearly 80 mg / g, and in 3 hours, its adsorption of bilirubin was 112 mg / g.

[0051] Figure 5 The bile acid adsorption kinetic curve of COF-A, where the dosage of the covalent organic framework material is 10 mg, the bile acid concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, COF-A adsorbs more than 90 mg / g of bile acid, and within 3 hours, its adsorption of bile acid is about 135 mg / g.

[0052] Figure 6 This is the albumin adsorption diagram of COF-A. It can be seen that the initial albumin solubility in the solution is 40 g / L. COF-A is added to make the concentration of COF-A 1 mg / mL. After 3 hours, the albumin solubility is 39.5 g / L, and the protein concentration is only reduced by 1.25%, indicating that COF-A adsorbs less albumin and has good selectivity.

[0053] Figure 7 This is the hemolysis rate spectrum of COF-A, where the concentration of COF-A is 1 mg / mL. It can be seen that the hemolysis rate of COF-A is only 0.6%. Example 2

[0054] 30 mg of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and 21 mg of 2,5-di-hydroxyterephthalaldehyde were added to a mixed solvent of 1.7 mL of mesitylene and 0.3 mL of dioxane, and then 0.2 mL of 8M acetic acid was added and ultrasonically dispersed for 10 minutes. After three freeze-pump-thaw cycles, the container was sealed and then heated at 120 o C, and reacted for 3 days in an undisturbed environment. As the reaction proceeded, a precipitate was gradually generated. The mother liquor was removed by centrifugation, and the precipitate was collected and washed with dimethylacetamide (DMAc) 3 times, then washed with deionized water 3 times, and finally replaced with anhydrous ethanol 3 times, and then vacuum dried at 150 °C for 24 h to obtain the covalent organic framework material COF-B.

[0055] Figure 8 is the X-ray powder diffraction pattern of COF-B, Figure 8 Curve 3 in the figure represents the simulation curve of COF-B, and curve 4 represents the measurement curve of the synthesized COF-B. The characteristic peak in curve 4 coincides with curve 3, indicating that the above covalent organic framework material has been successfully synthesized.

[0056] Fig. 9 is the infrared spectrum of COF-B, where 1640 cm -1The appearance of the characteristic peak of imine bond indicated that the monomers were covalently bound.

[0057] Fig.10 N of COF-B 2 The adsorption and desorption curves show that the adsorption isotherm curve presents type IV, and the surface COF-F has a mesoporous structure.

[0058] Fig.11 The kinetic curve of bilirubin adsorption of COF-B is shown in Figure 1, where the dosage of the covalent organic framework material is 10 mg, the bilirubin concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, the adsorption of bilirubin by COF-B is 107 mg / g, and within 3 hours, the adsorption of bilirubin is 154 mg / g.

[0059] Fig.12 The bile acid adsorption kinetic curve of COF-B, where the dosage of the covalent organic framework material is 10 mg, the bile acid concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, COF-B adsorbs 129 mg / g of bile acid, and within 3 hours, its adsorption of bile acid exceeds 185 mg / g.

[0060] Fig.13 This is the albumin adsorption diagram of COF-B. It can be seen that the initial albumin solubility in the solution is 40g / L. After adding COF-B for 3 hours, the albumin solubility is 39.6g / L, and the protein concentration is only reduced by 1%, indicating that COF-B adsorbs less albumin and has good selectivity.

[0061] Fig.14 This is the hemolysis rate spectrum of COF-B, where the concentration of COF-B is 1 mg / mL. It can be seen that the hemolysis rate of COF-B is only 0.9%. Example 3

[0062] 28 mg of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and 16 mg of terephthalaldehyde were added to a mixed solution of 0.5 mL of o-dichlorobenzene and 0.5 mL of 1-butanol. After three freeze-pump-thaw cycles, the container was sealed and then reacted at 120°C for 3 days without interference. As the reaction proceeded, a precipitate gradually formed. A yellow solid was produced, which was separated by filtration and washed with acetone in a Soxhlet extractor for 24 hours. The material was then activated with supercritical carbon dioxide, followed by dynamic vacuum drying at room temperature for 16 hours, and then dynamic vacuum drying at 120°C for 2 hours to obtain the covalent organic framework material COF-C.

[0063] Fig.15 is the X-ray powder diffraction pattern of COF-C, Fig.15 Curve 5 in the figure represents the simulation curve of COF-C, and curve 6 represents the measurement curve of the synthesized COF-C. The characteristic peak in curve 6 coincides with curve 5, indicating that the above covalent organic framework material has been successfully synthesized.

[0064] Fig.16 is the infrared spectrum of COF-C, where 1640 cm -1 The appearance of the characteristic peak of imine bond indicated that the monomers were covalently bound.

[0065] Fig.17 N of COF-C 2 The adsorption and desorption curves show that the adsorption isotherm curve presents type IV, and the surface COF-F has a mesoporous structure.

[0066] Fig.18 The kinetic curve of bilirubin adsorption of COF-C is shown in Figure 1, where the dosage of the covalent organic framework material is 10 mg, the bilirubin concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, the adsorption of bilirubin by COF-C is 48 mg / g, and within 3 hours, the adsorption of bilirubin is 86 mg / g.

[0067] Fig.19 The bile acid adsorption kinetic curve of COF-C is shown in Figure 1, where the dosage of the covalent organic framework material is 10 mg, the bile acid concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, the adsorption of bile acid by COF-C is 58 mg / g, and within 3 hours, the adsorption of bile acid is 103 mg / g.

[0068] Fig. 20 This is the albumin adsorption diagram of COF-C. It can be seen that the initial albumin solubility in the solution is 40g / L. After adding COF-C for 3 hours, the albumin solubility is 39.9g / L, and the protein concentration is only reduced by 0.25%, indicating that COF-C adsorbs less albumin and has good selectivity.

[0069] Fig.21 This is the hemolysis rate spectrum of COF-C, where the concentration of COF-C is 1 mg / mL. It can be seen that the hemolysis rate of COF-C is only 1.2%. Example 4

[0070] 14.1 mg of 1,3,5-tris(4-aminophenyl)benzene and 14.5 mg of 2,5-bis(prop-2-yn-1-yloxy)terephthalaldehyde were added to 5 mL of acetonitrile solution and dispersed by ultrasound. Then 0.5 mL of 12 M acetic acid was added, shaken and left at room temperature for 3 days. As the reaction proceeded, a precipitate gradually formed. The mother liquor was removed by centrifugation, and the precipitate was collected and washed 5 times with tetrahydrofuran (THF), replaced 5 times with anhydrous ethanol, and then passed through supercritical CO 2 After drying, the covalent organic framework material COF-D was obtained.

[0071] Fig. 22 is the X-ray powder diffraction pattern of COF-D, Fig. 22 Curve 7 in the figure represents the simulation curve of COF-D, and curve 8 represents the measurement curve of the synthesized COF-D. The characteristic peak in curve 8 coincides with curve 7, indicating that the above covalent organic framework material has been successfully synthesized.

[0072] Fig.23 is the infrared spectrum of COF-D, where 1640 cm -1 The appearance of the characteristic peak of imine bond indicated that the monomers were covalently bound.

[0073] Fig.24 N of COF-D 2 The adsorption and desorption curves show that the adsorption isotherm curve presents type IV, and the surface COF-F has a mesoporous structure.

[0074] Fig.25 The kinetic curve of bilirubin adsorption of COF-D is shown in Figure 1, where the dosage of the covalent organic framework material is 10 mg, the bilirubin concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, the adsorption of bilirubin by COF-D is 71 mg / g, and within 3 hours, the adsorption of bilirubin is 103 mg / g.

[0075] Fig.26 The bile acid adsorption kinetic curve of COF-D is shown in Figure 1, where the dosage of the covalent organic framework material is 10 mg, the bile acid concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, COF-D adsorbs 85 mg / g of bile acid, and within 3 hours, its adsorption of bile acid is 123 mg / g.

[0076] Fig. 27Albumin adsorption diagram of COF-D. It can be seen that the initial albumin concentration in the solution is 40 g / L. After adding COF-D for 3 hours, the albumin concentration is 39.8 g / L, and the protein concentration only decreases by 0.5%, indicating that COF-D has less adsorption of albumin and has good selectivity.

[0077] Fig.28 Hemolysis rate spectrum of COF-D, where the concentration of COF-D is 1 mg / mL. It can be seen that the hemolysis rate of COF-D is only 0.8%. Example 5

[0078] Add 11.3 mg of 1,3,6,8-tetrakis(4-aminophenyl)pyrene and 6.6 mg of 2,5-dihydroxyterephthalaldehyde to a mixed solvent of mesitylene and dioxane with a ratio of 1:1, with a total volume of 1 mL, and ultrasonically disperse. Then add 0.1 mL of 6 M acetic acid and shake well. After 3 freeze-pump thaw cycles, seal the container and then react at 120 o °C in a non-interfering environment for 3 days. As the reaction proceeds, precipitation gradually forms. Centrifuge to remove the mother liquor, collect the precipitate, wash it 3 times with tetrahydrofuran (THF), and then displace it 3 times with absolute ethanol. Subsequently, dry it under vacuum at 120 o °C for 24 h to obtain the covalent organic framework material COF-E.

[0079] Fig.29 X-ray powder diffraction pattern of COF-E Fig.29 Curve 9 in it represents the simulated curve of COF-E, and curve 10 represents the measured curve of the synthesized COF-E. The characteristic peaks in curve 10 coincide with curve 9, indicating the successful synthesis of the above covalent organic framework material.

[0080] Fig.30 Infrared spectrum of COF-E, where the appearance of the characteristic peak of the imine bond at 1640 cm -1 indicates the covalent binding of the monomers.

[0081] Fig.31 N 2 adsorption and desorption curve spectrum of COF-E. The adsorption isotherm curve shows type IV, indicating that COF-F has a mesoporous structure.

[0082] Fig.32The kinetic curve of bilirubin adsorption of COF-E, where the dosage of the covalent organic framework material is 10 mg, the bilirubin concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, the adsorption of bilirubin by COF-E is 85 mg / g, and within 3 hours, the adsorption of bilirubin is 140 mg / g.

[0083] Fig.33 The bile acid adsorption kinetic curve of COF-E, where the dosage of the covalent organic framework material is 10 mg, the bile acid concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, COF-E adsorbs 101 mg / g of bile acid, and within 3 hours, its adsorption of bile acid is 168 mg / g.

[0084] Fig.34 This is the albumin adsorption diagram of COF-E. It can be seen that the initial albumin solubility in the solution is 40g / L. After adding COF-E for 3 hours, the albumin solubility is 39.9g / L, and the protein concentration is only reduced by 0.25%, indicating that COF-E adsorbs less albumin and has good selectivity.

[0085] Fig.35 This is the hemolysis rate spectrum of COF-E, where the concentration of COF-E is 1 mg / mL. It can be seen that the hemolysis rate of COF-E is only 0.2%. Example 6

[0086] 60 mg of tetrakis-(4-aminophenyl)ethylene and 41 mg of terephthalaldehyde were added to 2 mL of dioxane and dispersed by ultrasound. 0.2 mL of 6 M acetic acid was added and shaken. After three freeze-pump-thaw cycles, the container was sealed and then heated at 120 o C for 3 days in an undisturbed environment. As the reaction proceeds, a precipitate is gradually generated. The mother liquor is removed by centrifugation, and the precipitate is collected and washed 3 times with tetrahydrofuran (THF), replaced 3 times with anhydrous ethanol, and then heated at 120 o C and vacuum dried for 24 h to obtain the covalent organic framework material COF-F.

[0087] Fig.36 is the X-ray powder diffraction pattern of COF-F, Fig.36 Curve 11 in the figure represents the simulation curve of COF-F, and curve 12 represents the measurement curve of the synthesized COF-F. The characteristic peak in curve 12 coincides with curve 11, indicating that the above covalent organic framework material has been successfully synthesized.

[0088] Fig.37is the infrared spectrum of COF-F, where 1640 cm -1 The appearance of the characteristic peak of imine bond indicated that the monomers were covalently bound.

[0089] Fig.38 N of COF-F 2 The adsorption and desorption curves show that the adsorption isotherm curve presents type IV, and the surface COF-F has a mesoporous structure.

[0090] Fig.39 The kinetic curve of bilirubin adsorption of COF-F is shown in Figure 1, where the dosage of the covalent organic framework material is 10 mg, the bilirubin concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, the adsorption of bilirubin by COF-F is 58 mg / g, and within 3 hours, the adsorption of bilirubin is 126 mg / g.

[0091] Fig.40 The bile acid adsorption kinetic curve of COF-F is shown in the figure, where the dosage of the covalent organic framework material is 10 mg, the bile acid concentration is 200 mg / L, the albumin concentration is 40 g / L, and the oscillation frequency is 200 rpm. It can be seen that within 30 minutes, the adsorption of bile acid by COF-F is 70 mg / g, and within 3 hours, the adsorption of bile acid is 151 mg / g.

[0092] Fig.41 This is the albumin adsorption diagram of COF-F. It can be seen that the initial albumin solubility in the solution is 40g / L. After adding COF-F for 3 hours, the albumin solubility is 39.7g / L, and the protein concentration is only reduced by 0.75%, indicating that COF-F adsorbs less albumin and has good selectivity.

[0093] Fig.42 This is the hemolysis rate spectrum of COF-F, where the concentration of COF-F is 1 mg / mL. It can be seen that the hemolysis rate of COF-F is only 0.4%.

[0094] In addition, for the covalent organic framework materials in Examples 1-6, 10 mg of the covalent organic framework material was brought into contact with 10 mL of plasma from patients with liver disease or blood from normal subjects, and then tested. The results are shown in Tables 1 and 2, wherein Table 1 shows the results of blood biochemical examinations of the plasma from patients with liver disease after contact with the covalent organic framework material, and Table 2 shows the results of routine blood examinations of normal subjects after contact with the covalent organic framework material.

[0095] Table 1

[0096] Table 2

[0097] It can be seen from Tables 1 and 2 that after contact with the covalent organic framework material, the content of direct bilirubin and total bilirubin is greatly reduced, while the content of other substances remains basically unchanged. It can be seen that the covalent organic framework material can adsorb harmful substances in the patient's blood and has excellent selectivity.

[0098] It can be seen from the above examples that the covalent organic framework material has an excellent adsorption effect on protein-bound toxins such as bilirubin and bile acid in the blood and large molecular toxins, and also has the characteristics of excellent adsorption selectivity, low hemolysis rate, and anti-protein adhesion. It is an excellent material as a blood perfusion adsorbent and has broad market applications.

Claims

1. Use of covalent organic framework materials in the preparation of blood perfusion adsorbents.

2. The use according to claim 1, It is characterized in that The covalent organic framework material is formed by reacting a first monomer containing an amino group and a second monomer containing an aldehyde group.

3. The use according to claim 2, It is characterized in that The first monomer is 1,3,5-tris(4-aminophenyl)benzene, tetrakis(4-aminophenyl)ethylene, 1,3,6,8-tetrakis(4-aminophenyl)pyrene or 2,4,6-tris(4-aminophenyl)-1,3,5-triazine.

4. The use according to claim 2, It is characterized in that The second monomer is phthalaldehyde, 2,5-di-hydroxyterephthalaldehyde or 2,5-bis(prop-2-yn-1-yloxy)terephthalaldehyde.

5. The use according to claim 2, It is characterized in that The covalent organic framework material is one or more of the following: The covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and terephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is shown in FIG1 ; The covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and 2,5-di-hydroxyterephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is shown in FIG8 ; The covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)-1,3,5-triazine and terephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is shown in FIG15 ; The covalent organic framework material is formed by the reaction of 2,4,6-tris(4-aminophenyl)benzene and 2,5-bis(prop-2-yn-1-yloxy)terephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is shown in FIG22 ; The covalent organic framework material is formed by the reaction of 1,3,6,8-tetrakis(4-aminophenyl)pyrene and 2,5-di-hydroxyterephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is shown in FIG29 ; The covalent organic framework material is formed by the reaction of tetrakis-(4-aminophenyl)ethylene and terephthalaldehyde, and the X-ray powder diffraction pattern of the formed covalent organic framework material is shown in Figure 36.

6. The use according to claim 1, It is characterized in that The target adsorbent of the blood perfusion adsorbent is protein-bound toxins and / or macromolecular toxins in the patient's blood.

7. The use according to claim 6, It is characterized in that The target adsorbed substance is bilirubin and / or bile acid in the patient's blood.

8. The use according to claim 1, It is characterized in that The pore size of the covalent organic framework material is larger than the diameter of the target adsorbent of the hemoperfusion adsorbent.

9. The use according to any one of claims 1 to 8, It is characterized in that The covalent organic framework material is used in combination with a second adsorption material, and the second adsorption material is selected from one or more of the group consisting of activated carbon, polysaccharides, resins, and immunosorbents.

10. The use according to claim 9, It is characterized in that The immunosorbent is an antigen-antibody binding adsorbent, a complement binding adsorbent, an Fc binding adsorbent, an electrostatic binding adsorbent or a hydrophobic binding adsorbent.

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