Cell collection material and cell collection column

Abstract

To provide an immunosuppressive cell collecting material whose performance to selectively collect immunosuppressive cells is improved, and an immunosuppressive cell collecting column with which the cell collecting material is filled.SOLUTION: An immunosuppressive cell collecting material is provided on the surface with polymer having a substituent group expressed in a following general formula (1) in an aromatic residue group of a main chain and is composed of a polymer molded product, and an immunosuppressive cell collecting column is filled with the cell collecting material. φ-R-X-Y-Z (1)[in the formula, φ shows an aromatic nucleus of a main chain, R shows -CH2-, -CH2 NHCOCH2-, or -CH2 NHCOCH2CH2-, X shows -NH(CH2)n-A-(CH2)mNH-, n and m show 2 or 3 in the same way or differently, A shows -NH- or -NH-(CH2CH2)p-NH-, p shows integers 2 to 4, Y shows a carbonyl group, -CH2CONH- or -CH2CH2CONH- group, and Z shows a linear or branched-chain alkyl group with 3 to 8 carbon number.]SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an immunosuppressive cell collection material primarily used for medical purposes, and an immunosuppressive cell collection column packed with said collection material. [Background technology]

[0002] Even in today's world of advanced medicine, new infectious diseases caused by new viruses and bacteria continue to emerge, threatening people's lives. Furthermore, cancer accounts for more than 300,000 deaths per year. Strengthening the immune system is essential to overcoming infectious diseases and cancer. Meanwhile, autoimmune diseases caused by an overactive immune system, in which immune cells attack the body's own tissues, are also on the rise. Subclasses of white blood cells that play an important role in maintaining immunity include killer T cells (CTLs) and natural killer cells (NK cells), which eliminate pathogens and viruses, B cells that produce antibodies, and helper T cells, which assist the functions of these cells. Regulatory T cells, regulatory B cells, and immunosuppressive granulocytes are also known to suppress runaway immune responses. Regulatory T cells include CD4 T cells, which express the transcription factor Foxp3 within their cells. + There are T cells (Treg) (Non-patent Document 1). There are also LAP cells that have latency associated peptide (LAP) on their cell surface. + CD4 + T cells, LAP + CD8 + T cells, LAP + B cells, etc. Regarding the relationship between Foxp3 and LAP, it has been reported that activation of Treg results in expression of LAP on the cell surface (Non-Patent Document 2).

[0003] The present inventors have been developing an adsorbent for LAP-positive cells, believing that if an adsorbent with high selectivity for immunosuppressive cells could be developed, it could be used to treat cancer, infectious diseases, and autoimmune diseases. They have found that extracorporeal circulation treatment of cancer-bearing rats using a column packed with fibers coated with polysulfone having diethylenetriamine groups as an adsorption ligand results in the inhibition of tumor growth and the extension of survival time (Patent Document 1). Furthermore, they have found that a column packed with fibers coated with polysulfone having leucine residue-linked diethylenetriamine groups has improved adsorption for LAP-positive cells and is highly effective in saving the lives of sepsis model rats (Patent Document 2).

[0004] Adaxacolumn is an adsorbent material that adsorbs immune-related cells. It is an adsorbent for activated granulocytes, and its ability to adsorb immunosuppressive lymphocytes has not been reported. Another related treatment technology is the polymyxin B-immobilized fiber column "Toraymyxin," which has been put to practical use as an extracorporeal circulation column for treating sepsis. Because this column has the ability to adsorb endotoxin, it is thought to be effective in suppressing endotoxin-induced inflammation in the early stages of sepsis. However, according to the results of a multicenter randomized controlled clinical trial conducted in the United States, the survival rate up to 28 days, which accounts for the majority of immunocompromised patients, was not significantly different from that of conventional drug treatments, suggesting that this treatment is less effective in the later stages of sepsis when the immune system is weakened (Non-Patent Document 3).

[0005] Recently, based on the idea that deaths from sepsis are caused by a weakened immune system, checkpoint antibody drugs such as anti-PD-1 antibodies, which were first developed as treatments for cancers such as melanoma, have begun to be tried in the treatment of sepsis. However, there are concerns about the occurrence of side effects such as the onset of autoimmune diseases.

[0006] Additionally, enterotoxin adsorption columns using mannose-binding lectin as a ligand have been proposed for the treatment of gram-positive bacterial infections. These are used for the purpose of removing toxins. Furthermore, systemic inflammatory response syndrome (SIRS) is a severe condition caused by an abnormal increase in inflammatory cytokines induced by an infection. Since histones are one of the causative agents, histone removal columns have been proposed for treatment (Patent Document 3). Although numerous methods have been proposed for removing the liquid causative agents that cause the disease, it is difficult to say that they have provided sufficient therapeutic effects.

[0007] Therefore, we have proposed a more effective method for directly removing immunosuppressive cells. Until now, there have been no effective adsorbents, except for columns in which antibodies are immobilized on beads or other materials as adsorption ligands. Compared to adsorbents with immobilized antibodies, adsorbents that use artificial substances as adsorption ligands have the advantage of being able to be sterilized by high-pressure steam, making them suitable for use as medical materials, but they have the disadvantage of low adsorption selectivity. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Patent No. 5916712 [Patent Document 2] Japanese Patent Application Publication No. 2019-205552 [Patent Document 3] International Publication No. 2016 / 013540 [Non-patent literature]

[0009] [Non-Patent Document 1] M. Beyer and JL Schultze, Blood 2006: 108: 804-811 [Non-patent document 2] K. Nakamura, A. Kitani, I. Fuss, A. Pedersen, N. Harada, H. Nawata and W. Strober, The Journal of Immunology 2004: 172: 834-842 [Non-patent document 3] R. Phillip Dellinger at al, Journal American Medical Association 2018: 320(14): 1455-1463. Summary of the Invention [Problem to be solved by the invention]

[0010] An object of the present invention is to provide an immunosuppressive cell collection material with improved performance for selectively capturing immunosuppressive cells, and an immunosuppressive cell collection column packed with the collection material. [Means for solving the problem]

[0011] The present inventors have conducted extensive research to develop a ligand that can achieve selectivity comparable to that of antibodies, and have found that antibody-level selectivity can be achieved by optimizing the structure of the amino group and the alkyl group at the end of the substituent. Specifically, they have discovered that immunosuppressive cells can be remarkably selectively captured by using a linear aromatic polymer having a polyamine-containing linking group to which an alkyl group having 3 to 8 carbon atoms is bonded.

[0012] The present invention was completed based on these findings and through further investigation, and provides the following immunosuppressive cell capture material and immunosuppressive cell capture column.

[0013] (I) Immunosuppressive cell collection material (I-1) An immunosuppressive cell collection material comprising a molded body having, on its surface, an aromatic polymer having a substituent represented by the following general formula (1) in the aromatic residue of the main chain. φ-RXYZ (1) [wherein φ represents an aromatic nucleus in the main chain, R represents -CH2-, -CH2NHCOCH2-, or -CH2NHCOCH2CH2-; X is -NH(CH2) n -A-(CH2) m represents -NH-, n and m are the same or different and represent 2 or 3, and A represents -NH- or -NH-(CH2CH2) p -NH-, and p is an integer of 2 to 4; Y represents a carbonyl group, -CHCONH- or -CHCHCONH- group; Z represents a linear or branched alkyl group having 3 to 8 carbon atoms. (I-2) The capturing material according to (I-1), in which the substituents represented by the general formula (1) are bonded at a frequency of 1 to 50 per 100 aromatic nuclei. (I-3) The adsorption material according to (I-1) or (I-2), wherein X is a diethylenetriamine residue, a bis(3-aminopropyl)amine residue, or a 1,5,8,12-tetraazadodecane residue. (I-4) The trapping material according to any one of (I-1) to (I-3), wherein Z is an isoamyl group or a pentyl group. (I-5) The collection material according to any one of (I-1) to (I-4), wherein the aromatic polymer is polysulfone, polyetherimide, polyimide, or a derivative thereof. (I-6) The capturing material according to any one of (I-1) to (I-5), which is a molded product obtained by applying the aromatic polymer to a fiber. (I-7) The collection material according to any one of (I-1) to (I-6), wherein the immunosuppressive cells are lymphocytes having latency-associated protein on the cell surface.

[0014] (II) Immunosuppressive cell collection column (II-1) A column for collecting immunosuppressive cells, which is packed with the collection material according to any one of (I-1) to (I-7). (II-2) The column according to (II-1), which is for extracorporeal circulation. (II-3) The column according to (II-1), which is used for cell therapy. (II-4) The column according to any one of (II-1) to (II-3), which is used for treating an infectious disease, an opportunistic infection, a burn, preventing cancer recurrence after cancer removal surgery, treating cancer, or preventing sepsis. [Effects of the Invention]

[0015] The capture material and column of the present invention can selectively capture immunosuppressive cells from blood, reducing their concentration and enhancing immunity. Therefore, the capture material and column of the present invention are expected to be applied to the treatment of severe infections and cancer.

[0016] Furthermore, the collection material of the present invention uses a non-proteinaceous material, so that it can be sterilized. DETAILED DESCRIPTION OF THE INVENTION

[0017] Hereinafter, the embodiments of the present invention will be described in more detail.

[0018] Immunosuppressive cell collection material The immunosuppressive cell capture material of the present invention is characterized by comprising a molded body having, on its surface, an aromatic polymer having a substituent represented by the following general formula (1) in the aromatic residue of the main chain. φ-RXYZ (1) [wherein φ represents an aromatic nucleus in the main chain, R represents -CH2-, -CH2NHCOCH2-, or -CH2NHCOCH2CH2-; X is -NH(CH2) n -A-(CH2) m represents -NH-, n and m are the same or different and represent 2 or 3, and A represents -NH- or -NH-(CH2CH2) p -NH-, and p is an integer of 2 to 4; Y represents a carbonyl group, -CHCONH- or -CHCHCONH- group; Z represents a linear or branched alkyl group having 3 to 8 carbon atoms.

[0019] The immunosuppressive cell capture material of the present invention preferably has groups represented by general formula (1) bonded as aromatic nucleus substituents at a frequency of 1 to 50 per 100 aromatic nuclei.

[0020] (Composition of collection material) The immunosuppressive cells to be collected in the present invention are cells having LAP on their cell surface. + CD4 + Regulatory T cells and LAP + CD8 + Examples of LAP-positive cells include regulatory T cells, LAP-positive B cells, LAP-positive monocytes and granulocytes. The presence of these cells can be confirmed by flow cytometry analysis. + CD4 + Regulatory T cells and LAP + CD8 + The abundance of regulatory T cells in peripheral blood varies depending on the individual, but generally accounts for 5% to 30% of each T cell subset. The proportion of LAP-positive B cells ranges from 5% to 60%.

[0021] The collection material of the present invention has high selective adsorption for LAP-positive lymphocytes. + It selectively adsorbs T cells. The degree of selectivity for LAP-positive cells is higher than that of CD4 + T cells and CD8 + T cells are at the same level, and B cells are slightly lower compared to these.

[0022] In general formula (1), Z is a linear or branched alkyl group having 3 to 8 carbon atoms, specifically, isobutyl, n-pentyl, isoamyl, hexyl, heptyl, octyl, etc., and is preferably an isoamyl or pentyl group. Branched alkyl groups have higher selectivity, and isobutyl groups have higher selectivity than butyl groups. Ethyl groups with short chain lengths decrease selectivity. On the other hand, decanoyl groups with long chain lengths decrease LAP-positive cell adsorption. Another drawback is that longer alkyl chains increase the platelet capture rate.

[0023] X in general formula (1) is a linear polyamine chain and must have two or more basic amino groups. Too many basic amino groups increases the adsorption of plasma proteins such as albumin and reduces the selectivity of cell adsorption, so two to four basic amino groups are preferred. Specific examples of X include a diethylenetriamine residue, a bis(3-aminopropyl)amine residue, and a bis(3-aminopropyl)ethylenediamine residue.

[0024] In the present invention, "selectively capture" means that when blood is passed through a column packed with a collection material, the proportion of immunosuppressive cells in the passing blood decreases compared to before the passage, and the proportion of immunosuppressive cells in the captured cells increases compared to before the passage.

[0025] Higher cell selectivity is preferable from the perspective of medical safety, but to improve cell selectivity, lower amino group basicity and lower presence density are generally better. Furthermore, in applications where blood is directly processed and returned to the living body, such as extracorporeal circulation, it is particularly important to avoid, for medical safety reasons, the amino groups of the adsorption material stimulating cells captured by the adsorption material, causing the release of inflammatory cytokines or inducing necrosis in the adsorbed cells. From these perspectives, the amount of amino groups in the molded article of the present invention is preferably 150 μmol or less, 110 μmol or less, and more preferably 1 to 110 μmol per gram of the molded article.

[0026] In the present invention, the aromatic polymer refers to an aromatic polymer with a linear polymer backbone that can be molded. Furthermore, polymers that can withstand gamma ray sterilization and high-pressure steam sterilization are particularly preferred. Solubility in organic solvents is preferred because it can be applied to the surface of other molded products. Film-forming properties are particularly preferred from a safety perspective, as they increase the mechanical stability of the processed product and reduce the probability of fine particles peeling off from the molded product. A specific example of the polymer is polysulfone-{(p-CH)-SO-(p-CH)-O-(p-CH)-C(CH)-(p-CH)-O}, which is composed of bisphenol A and diphenyl sulfone. n-, poly(p-phenylene ether sulfone)-{(p-C6H4)-SO2-(p-C6H4)-O-(p-C6H4)-O} n -, -{(p-C6H4)-SO2-(p-C6H4)-O-(p-C6H4)-C(CF3)2-(p-C6H4)-O} n Examples of suitable polymers include aromatic polysulfone polymers, polyetherimides, polyimides, and derivatives thereof, such as those represented by the above-mentioned olefins. Among these, polysulfones are particularly preferred because they are inexpensive, highly processable, possess high mechanical strength, are soluble in a variety of organic solvents, and are capable of forming tough films, making them excellent medical materials. Furthermore, functional groups can be easily introduced into them. Furthermore, among the above-mentioned polymers, those that dissolve in non-chlorine-based, low-toxicity solvents such as tetrahydrofuran, dimethyl sulfoxide, N,N-dimethylacetamide, and N-methylpyrrolidone are particularly preferred from the perspective of preventing environmental pollution and maintaining occupational health. The molecular weight of the polymer is not particularly limited as long as it can be molded; however, a molecular weight of typically 10,000 to 5,000,000, and particularly 20,000 to 200,000, is preferred for good moldability.

[0027] The shape of the molded article of the present invention is not particularly limited, and examples thereof include fibers, nonwoven fabrics, membranes, hollow fibers, granules, and advanced processed products thereof, and the shape is appropriately selected depending on the application. Examples of the molded article of the present invention include a molded product of the aromatic polymer itself, and a product in which the aromatic polymer is coated on the surface of another molded article.

[0028] The shape of the molded product may be fiber, nonwoven fabric, membrane, hollow fiber, powder, granule, or any of these advanced processed products. Materials for the molded product include polyester, polyamide, polyurethane, and the like. Specific examples of polyester include polylactic acid, polyglycolic acid, polyethylene terephthalate, and polybutylene terephthalate. Specific examples of polyamide include nylon-6 and polyhexamethylene adipamide. The molecular weight of such polymers is not particularly limited as long as it is within a moldable range.

[0029] The capture material of the present invention can selectively capture immunosuppressive cells from a cell fluid containing many types of blood cells. Furthermore, because the capture material of the present invention does not contain protein components, it does not lose its function during sterilization procedures such as high-pressure steam sterilization and radiation sterilization.

[0030] (Method of manufacturing the collection material) The simplest example of a method for producing the immunosuppressive cell capture material of the present invention is a method in which an HXYZ molecule is reacted with an aromatic polymer having a reactive functional group. The HXYZ molecule can be obtained by reacting an HXH molecule with a Cl-YZ molecule. Either reaction can be carried out at a temperature between room temperature and 100°C.

[0031] Examples of aromatic polymers with reactive functional groups include polymers with reactive functional groups such as chloromethyl groups, haloacetamidomethyl groups, and haloacetamidoethyl groups as aromatic nucleus substituents. Polymers with chloroacetamidomethyl groups are particularly preferred due to their balance of high reactivity and stability during processing. These polymers can be easily synthesized by adding a sulfuric acid solution of N-hydroxymethyl-2-chloroacetamide to a nitrobenzene solution of polysulfone. The chloro groups can be easily converted to iodo groups by mixing with an ethanol solution of potassium iodide.

[0032] Furthermore, an aromatic polymer having a substituent represented by general formula (1) can be produced by adding a solution of HXYZ molecules to a solution or molded product of a polymer such as polysulfone having reactive functional groups such as chloromethyl groups, haloacetamidomethyl groups, haloacetamidoethyl groups, etc. The adsorbent can be produced by molding this polymer into fibers or coating the surface of a molded product.

[0033] The method of coating with a polymer can be a method of immersing the molded article in a polymer solution and then evaporating off the solvent. Specific examples are shown below, but the present invention is not limited to these.

[0034] First, polysulfone is reacted with 0.5 times the molar amount of N-hydroxymethyl-2-chloroacetamide to prepare chloroacetamidomethyl polysulfone with a substitution rate of 50 mol%. Next, this polymer is dissolved in dimethylacetamide to prepare a polymer solution. A polyethylene terephthalate nonwoven fabric is immersed in this solution, and the dimethylacetamide is evaporated under reduced pressure to obtain a polymer-coated nonwoven fabric. The polymer-coated nonwoven fabric is then immersed in a solvent containing a ligand molecule with the chemical structure HXYZ at 20-80°C for 1-30 hours to obtain the adsorbent of the present invention. The solvent is not particularly limited as long as it does not react with the coated polymer or the ligand molecule and does not dissolve the coated polymer, but dissolves the ligand molecule. Typically, water, methanol, ethanol, and mixtures thereof are preferred due to their low toxicity and ease of handling. Alternatively, the adsorbent of the present invention can be obtained by immersing the polymer-coated nonwoven fabric in a solvent containing a polyamine with a chemical structure of HXH for 1-30 hours at 20-80°C to bind the polyamine, rinsing with water, and then immersing the fabric in a solvent dissolving a ligand molecule with a chemical structure of Cl-CHCONH-Z or CH=CHCONH-Z for 1-30 hours at 20-80°C. Typically, water, methanol, ethanol, and mixtures thereof are preferably used as the solvent.

[0035] The amount of the coating polymer is preferably 0.1 to 50% by mass, more preferably 5 to 20% by mass, based on the molded product.

[0036] Immunosuppressive cell collection column The immunosuppressive cell collection column of the present invention is characterized by being packed with the immunosuppressive cell collection material described above.

[0037] When the adsorbent of the present invention is in the form of a fiber or nonwoven fabric, the packing density of the adsorbent of the present invention packed in the column is preferably 30 to 1,000 mg / cm. 3 , particularly preferably 150 to 600 mg / cm 3Within this range, cell collection efficiency, selectivity, and pressure drop during fluid flow are optimal. High pressure drop leads to hemolysis of red blood cells. Hemolysis is particularly contraindicated in extracorporeal circulation. Pressure drop is positively correlated with packing density and blood flow rate. To complete extracorporeal circulation in humans within two hours, the blood flow rate must be at least 30 mL / min, so pressure drop must be kept below 100 mmHg to prevent hemolysis of red blood cells.

[0038] The column of the present invention, when used as a column for extracorporeal circulation, can selectively collect immunosuppressive cells from blood and reduce their concentration. Furthermore, since leukocytes prepared from blood treated with the column of the present invention and leukocytes obtained by treating previously isolated leukocytes with the column have improved antitumor activity, the column of the present invention can also be used as a column for cell therapy for diseases such as cancer.

[0039] Immunosuppressive cells can be obtained by detaching cells adsorbed to the column of the present invention, and by expanding these cells in vitro, concentrated immunosuppressive cells can be obtained, which can be used in cell therapy for autoimmune diseases. Conventional immunosuppressive cell recovery methods using flow cytometers or magnetic beads involve binding antibodies to cells, but the cells recovered from the column in the present invention are characterized by being intact cells with no antibody bound thereto.

[0040] The container into which the adsorption material of the present invention is filled may be made of, for example, glass, plastic, stainless steel, etc., and the size of the container may be selected appropriately depending on the purpose of use.

[0041] Because the column of the present invention can selectively capture immunosuppressive cells, it can be used to prevent infections (particularly severe infections and opportunistic infections) caused by immunosuppression, bacterial infections resistant to antibiotics, infections (due to severe trauma, surgery, burns, etc.), and sepsis (after severe trauma). It can also be used to treat cancer and prevent cancer recurrence after cancer removal surgery. When the column of the present invention is used for cancer treatment, its combined use with surgical therapy, radiation therapy, anticancer drug therapy, activated leukocyte therapy, vaccine therapy, etc. is thought to be useful in improving the efficacy of these therapies and in preventing metastasis and recurrence in particular. Because the column of the present invention can be used for extracorporeal circulation even in a state of shock, it can be said to be a safe column.

[0042] The types of cancer to which the column of the present invention can be applied include stomach cancer, colorectal cancer (rectal cancer, colon cancer), small intestine cancer, liver cancer, pancreatic cancer, lung cancer, pharyngeal cancer, esophageal cancer, kidney cancer, gallbladder and bile duct cancer, head and neck cancer, skin cancer, bladder cancer, prostate cancer, breast cancer, uterine cancer (cervical cancer, endometrial cancer), ovarian cancer, brain tumor, thymoma, leukemia, malignant lymphoma, etc. [Example]

[0043] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0044] In the present Examples, the measurement of amino group amount, analysis of cell surface antigens, preparation of septic rats, extracorporeal circulation of rats, and in vitro adsorption tests were carried out by the following methods unless otherwise specified.

[0045] 1. Measurement of picric acid adsorption amount A 0.1 g sample was immersed in 10 mL of a 0.1 M picric acid / 70% ethanol solution and gently shaken for 2 hours. The sample was then washed with 70% ethanol until the yellow color of the wash disappeared. The sample was then immersed in 10-50 mL of a 1 wt% diethylamine / 70% ethanol solution to elute the picric acid. The picric acid concentration was determined from the absorbance at 320 nm, and the adsorbed amount was divided by the exact sample weight to determine the amount of picric acid adsorbed per gram. This value corresponds to the amount of amino groups.

[0046] 2. Analysis of cell surface antigens Cell surface antigen analysis was performed using a Beckman Coulter CytoFlex S multi-laser, multicolor flow cytometer. Seven to nine colors were used for multicolor measurement. The following antibodies were used for cell surface staining: fluorescein isothiocyanate (FITC)-conjugated anti-rat granulocyte marker (HIS48) antibody, phycoerythrin-cyanine 7 (PE-Cy7)-conjugated anti-rat CD4 antibody (e-Bioscience); peridinin-chlorophyll-protein-cyanine 5.5 (PerCP-Cy5.5)-conjugated anti-rat CD45RA (Biolegend); phycoerythrin (PE)-conjugated anti-rat CD3 antibody, allophycocyanin (APC)-conjugated anti-mouse LAP antibody, Alexa Fluor 700-conjugated anti-rat CD45 antibody, and V450-conjugated anti-rat CD8a antibody (Becton Dickinson).

[0047] 100 μL of blood was collected in a 15 mL test tube, and 100 μL of 10% mouse serum in Fax buffer (commercially available Dulbecco's phosphate buffer containing 0.1% bovine serum albumin) containing fluorescent antibodies was added. The tube was then left to stand at room temperature for 30 minutes. Next, 1 mL of Becton Dickinson hemolysate was added to the resulting cell pellet after centrifugation, and the tube was shaken for 20 minutes. After washing with Fax buffer, the cells were dispersed in 200 μL of Fax buffer and analyzed using a flow cytometer. A CD45 gate was applied, and 50,000 cells were collected, measured, and analyzed as follows.

[0048] The distribution of granulocytes and lymphocytes in the FCS (forward scatter)-SSC (side scatter) dot plot confirmed that hemolysis was successful. + CD3+CD8a+ cells were defined as T cells, and CD8 + T cells were identified as CD3-negative CD45RA-positive cells as B cells, and His-positive cells were identified as granulocytes. The FSC-SSC dot plots confirmed that the expressing cells matched.

[0049] 3. Preparation of Septic Rats Lipopolysaccharide (derived from Escherichia coli O111, phenol extract; Fujifilm Wako Pure Chemical Industries, Ltd. 125-05201; hereafter referred to as LPS) was dissolved in saline to a concentration of 10 mg / mL and sterilized through a 0.22 μm membrane (MILLEX®-GV filter unit) to prepare an LPS solution. Sepsis rats were prepared by intraperitoneally injecting 1 mL of LPS solution per kg body weight into male WKAH / Hkm rats (10-12 weeks old) 4 hours before extracorporeal circulation.

[0050] 4. Extracorporeal Circulation in Rats (Preparation of extracorporeal circulation column) An extracorporeal circulation column was prepared by packing 0.3 g of the collection material into a cylindrical polypropylene column with an inner diameter of 1 cm and an internal volume of 2 mL. The column and circuit were sterilized by passing 70% alcohol through them, and then pretreated by passing 40 mL of heparinized saline (5 units / mL) at a rate of 2 mL / min immediately before extracorporeal circulation.

[0051] (extracorporeal circulation) Rats weighing 350–450 g were anesthetized with a triple anesthetic mixture (7.5 mg of Domitor, 400 mg of midazolam "Sandoz," and 500 mg of Betolfar per liter of saline; 0.375 mg of Domitor per kg of rat body weight was injected subcutaneously). The left femoral artery and vein were cannulated. Blood was withdrawn from the artery, passed through an extracorporeal circulation column using a microtube pump, and returned to the vein. Extracorporeal circulation was performed for 30 minutes at a blood flow rate of 2 mL / min. During the extracorporeal circulation, heparin was continuously administered at 100 units / hour using an infusion pump (Terufusion Mini Syringe Pump TE-361N; Terumo Corporation). However, for anesthesia of septic rats, 90% of the usual dose of anesthetic was used.

[0052] After the end of extracorporeal circulation, a medetomidine antagonist (a solution containing 150 mg of antisedan per 1 L of physiological saline) was subcutaneously injected at a rate of 0.75 mg of antisedan per kg of rat body weight to wake the rat up.

[0053] 5. In vitro adsorption test (Evaluation of the adsorption ability of adsorbents to LAP-positive cells) Whole blood was collected from Donryu rats or WKAH / Hkm rats and used to evaluate the adsorbent. 0.1 g of adsorbent was steam sterilized in saline (120°C x 15 minutes). This adsorbent and 2 mL of rat blood were placed in a 50 mL polypropylene centrifuge tube and shaken at 37°C for 20 minutes. After adsorption, the blood was collected and surface antigens were measured using a flow cytometer.

[0054] [Manufacturing example] <Preparation of chloroacetamidomethylated polysulfone> (Preparation of Polymer 1) A mixed solution of 60 mL of nitrobenzene and 120 mL of sulfuric acid was cooled to 0°C, and then 13.6 g (0.11 mol) of N-hydroxymethyl-2-chloroacetamide was added and dissolved at 0-10°C. This solution was then added to a nitrobenzene solution (22.1 g: 0.05 mol / 200 mL) of polysulfone (Sigma-Aldrich-428302; average molecular weight: 35,000) with vigorous stirring. After stirring at 20°C for 2 hours, the reaction mixture was poured into a large excess of cold methanol to precipitate the polymer. The precipitate was extracted with methanol until the nitrobenzene odor disappeared and then dried to yield 24.3 g of polymer. This polymer was dissolved in 150 mL of dimethylacetamide and reprecipitated in a large excess of methanol. Since some of the chloroacetamidomethyl groups in this polymer had been hydrolyzed to aminomethyl groups, the following treatment was carried out to restore the chloroacetamidomethyl groups. Specifically, 24 g of the polymer was dissolved in 150 mL of dimethylacetamide, and 1 mL of chloroacetyl chloride was added dropwise over ice water with stirring. The reaction mixture was allowed to react for 20 hours. The reaction mixture was poured into a large excess of water to precipitate the polymer, which was then immersed in 2 L of water containing 3 g of sodium bicarbonate for 5 hours. This polymer was then dissolved in dimethylformamide, poured into methanol, and the polymer was precipitated again. The polymer was then dried in vacuo to obtain 27 g of coating polymer 1.

[0055] The polymer was soluble in dimethylformamide, dimethyl sulfoxide, methylene chloride, and tetrahydrofuran. Elemental analysis: N: 2.6% Cl: 6.1%. The infrared absorption spectrum showed peaks at 3290-3310, 1670, and 1528 cm -1 The presence of amide groups was confirmed by the absorption of deuterated chloroform solution. 1 HNMR spectrum was measured, and the substitution rate per repeating unit was confirmed to be 200% from the ratio of the area of ​​the peak (4.22 ppm) derived from the methylene group hydrogen (2H) of the benzyl group of the amidomethyl group to the area of ​​the peak (1.66 ppm; singlet) derived from the isopropylidene group hydrogen (6H) of the polysulfone main chain.

[0056] (Preparation of Polymer 2) A mixed solution of 30 mL of nitrobenzene and 60 mL of sulfuric acid was cooled to 0°C, and then 6.8 g (0.055 mol) of N-hydroxymethyl-2-chloroacetamide was added and dissolved at a temperature of 0-10°C. This solution was then added to a nitrobenzene solution (44.2 g: 0.1 mol / 1600 mL) of polysulfone (Sigma-Aldrich-428302; average molecular weight: 35,000) with vigorously stirring. After further stirring at 20°C for 2 hours, the reaction mixture was poured into a large excess of cold methanol to precipitate the polymer. The precipitate was extracted with methanol until the nitrobenzene odor disappeared and then dried to yield 48.1 g of polymer. 47 g of this polymer was dissolved in 300 mL of dimethylacetamide, and 1 mL of chloroacetyl chloride was added dropwise with stirring. The reaction mixture was then poured into a large excess of water to precipitate the polymer, which was then immersed in 2 L of water containing 3 g of sodium bicarbonate for 5 hours. The polymer was reprecipitated in a large excess of water to obtain 48 g of polymer 2. This polymer was soluble in dimethylformamide, dimethyl sulfoxide, and tetrahydrofuran. The presence of amide groups was confirmed by infrared absorption spectroscopy. 1 The 1H NMR spectrum confirmed that the substitution rate per repeating unit was 50%.

[0057] (Preparation of Polymer 3) A mixed solution of 15 mL of nitrobenzene and 30 mL of sulfuric acid was cooled to 0°C, and then 2.7 g (0.022 mol) of N-hydroxymethyl-2-chloroacetamide was added and dissolved at 0-10°C. This solution was then added to a nitrobenzene solution (44.2 g: 0.1 mol / 400 mL) of polysulfone (Sigma-Aldrich-428302, average molecular weight: 35,000) with vigorous stirring. After stirring at 20°C for 2 hours, the reaction mixture was poured into a large excess of cold methanol to precipitate the polymer. The precipitate was extracted with methanol until the nitrobenzene odor disappeared and then dried to obtain 44.0 g of polymer. 40 g of this polymer was dissolved in 300 mL of dimethylacetamide and poured into a large excess of methanol to precipitate the polymer, yielding 42 g of polymer 3. This polymer was dissolved in dimethylformamide, dimethyl sulfoxide, and tetrahydrofuran. The presence of amide groups was confirmed by infrared absorption spectroscopy. The deuterated chloroform solution 1 The 1H NMR spectrum confirmed that the substitution rate per repeating unit was 20%.

[0058] (Preparation of Polymer 4) A mixed solution of 15 mL of nitrobenzene and 30 mL of sulfuric acid was cooled to 0°C, and then 1.3 g (0.01 mol) of N-hydroxymethyl-2-chloroacetamide was added and dissolved at 0-10°C. This solution was added to a nitrobenzene solution (88.4 g: 0.2 mol / 800 mL) of polysulfone (Sigma-Aldrich-428302, average molecular weight: 35,000) with vigorous stirring. After stirring at 20°C for 2 hours, the reaction mixture was poured into a large excess of cold methanol to precipitate the polymer. The precipitate was extracted with methanol until the nitrobenzene odor disappeared and then dried to obtain 88.0 g of polymer. 40 g of this polymer was dissolved in 300 mL of dimethylacetamide and poured into a large excess of methanol to precipitate the polymer, yielding 39 g of polymer 4. This polymer was dissolved in dimethylformamide and tetrahydrofuran. The presence of amide groups was confirmed by infrared absorption spectroscopy. The deuterated chloroform solution 1The 1H NMR spectrum confirmed that the substitution rate per repeat unit was 5%.

[0059] <Preparation of adsorption material carrier> (Nonwoven fabric cleaning treatment) To remove foreign matter such as oil from the surface of the fibers, polyethylene terephthalate fiber nonwoven fabric (density 48 mg / cm 3 88.2 g of the nonwoven fabric (Nippon Vilene Co., Ltd.) was immersed in 2 L of a 0.5 wt% diethylenetriamine dimethyl sulfoxide solution and heated at 105°C for 20 minutes. After rinsing with water and drying, 87.2 g of the washed nonwoven fabric was obtained. The picric acid adsorption capacity was 1 μmol / g.

[0060] (Coated nonwoven fabric-1) 7.5 g of the polymer 4 prepared above was dissolved in 300 mL of tetrahydrofuran, and 50 g of the washed nonwoven fabric prepared above was immersed in this solution and allowed to stand for 24 hours. After that, the tetrahydrofuran was evaporated by rotating the flask, and then the mixture was dried in vacuum to obtain 57.5 g of coated nonwoven fabric-1.

[0061] (Coated nonwoven fabric-2) 28.3 g of the previously prepared coated nonwoven fabric-1 was immersed in 1 L of a 5% ethanol solution of diethylenetriamine and heated at 50°C for 2 hours. After rinsing with water and vacuum drying, 50 g of the previously prepared washed nonwoven fabric was immersed in a solution of 2.5 g of polymer 1 dissolved in 500 mL of dimethyl sulfoxide and allowed to stand for 24 hours. After draining, the nonwoven fabric was vacuum dried to obtain 29.8 g of coated nonwoven fabric-2.

[0062] (Coated nonwoven fabric-3) 4.5 g of the polymer 2 prepared previously was dissolved in 300 mL of dimethylacetamide, and 30.7 g of the washed nonwoven fabric prepared previously was immersed in this solution and allowed to stand for 24 hours. After that, the dimethylacetamide was evaporated under vacuum using a rotary evaporator while rotating the nonwoven fabric in a flask. The nonwoven fabric was then dried under vacuum to obtain 35.1 g of coated nonwoven fabric-3.

[0063] (Coated nonwoven fabric-4) 3.1 g of the polymer 3 prepared above was dissolved in 100 mL of methylene chloride, and 20 g of the washed nonwoven fabric prepared above was immersed in this solution and allowed to stand for 24 hours. The methylene chloride was evaporated by rotating the mixture in a flask, and then the mixture was dried in vacuum to obtain 23.1 g of coated nonwoven fabric 4.

[0064] (Coated nonwoven fabric-5) 3.1 g of the polymer 2 prepared above was dissolved in 100 mL of methylene chloride, and 20 g of the washed nonwoven fabric prepared above was immersed in this solution and allowed to stand for 24 hours. The methylene chloride was evaporated by rotating the mixture in a flask, and then the mixture was dried in vacuum to obtain 23.1 g of coated nonwoven fabric-5.

[0065] <Preparation of the present invention adsorption material-1 (isoamyl group type-1)> 4.8 g of coated nonwoven fabric-2 was immersed in a solution consisting of 2 g (9.6 mmol) of diethylenetriamino-acetyl-isoamylamide (prepared by reacting diethylenetriamine and N-isoamyl-chloroacetamide in an equimolar ratio), 4 mL of 2N NaOH, and 40 mL of 70% ethanol, and heated in a 50°C water bath for 7 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 4.6 g of nonwoven fabric (adsorbent material-1 of the present invention) was obtained, in which the ligand was diethylenetriamino-acetyl-isoamylamide groups (the aromatic nucleus substituent was 2,5,8,11,14-pentaaza-3,13-dioxo-17-methyloctadecanyl groups). The picric acid adsorption capacity was 45.2 μmol / g.

[0066] <Preparation of the present invention collection material-2 (isoamyl group type-2)> 4.6 g of coated nonwoven fabric-2 was immersed in a solution consisting of 1 g (9.6 mmol) of diethylenetriamino-acetyl-isoamylamide, 2 mL of 2N NaOH, and 40 mL of 50% dimethyl sulfoxide water, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 4.3 g of nonwoven fabric (adsorbent material-2 of the present invention) was obtained, in which the ligand was diethylenetriamino-acetyl-isoamylamide groups (the aromatic nucleus substituent was 2,5,8,11,14-pentaaza-3,13-dioxo-17-methyloctadecanyl groups). The picric acid adsorption capacity was 50.0 μmol / g.

[0067] <Preparation of the present invention adsorption material-3 (isoamyl group type-3)> 4.3 g of coated nonwoven fabric-1 was immersed in a solution consisting of 2 g (9.6 mmol) of diethylenetriamino-acetyl-isoamylamide, 4 mL of 2N NaOH, and 40 mL of 70% ethanol and heated in a 50°C water bath for 7 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 4.1 g of nonwoven fabric (inventive adsorption material-3) was obtained, in which the ligand was diethylenetriamino-acetyl-isoamylamide groups (the aromatic nucleus substituent was 2,5,8,11,14-pentaaza-3,13-dioxo-17-methyloctadecanyl groups). The picric acid adsorption capacity was 31.4 μmol / g.

[0068] <Preparation of the present invention adsorption material-4 (hexyl group type-1)> 3.1 g of coated nonwoven fabric-3 was immersed in a solution consisting of 1 g (3.7 mmol) of bis(3-aminopropyl)amine-acetyl-hexylamide (prepared by reacting bis(3-aminopropyl)amine with an equimolar amount of N-hexyl-chloroacetamide), 1 mL of 2N NaOH, and 40 mL of 70% ethanol, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 3.1 g of nonwoven fabric (inventive adsorption material-4) was obtained, in which the ligand was bis(3-aminopropyl)amine-acetyl-hexylamide (the aromatic nucleus substituent was 2,5,9,13,16-pentaaza-3,15-dioxo-docosanyl). The picric acid adsorption capacity was 10.7 μmol / g.

[0069] <Preparation of the present invention adsorption material-5 (octyl group type-1)> 3.2 g of coated nonwoven fabric-5 was immersed in a solution consisting of 1 g (3.3 mmol) of bis(3-aminopropyl)amine-acetyl-octylamide (prepared by reacting bis(3-aminopropyl)amine with an equimolar amount of N-octyl-chloroacetamide), 1 mL of 2N NaOH, and 40 mL of 70% ethanol, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 3.1 g of nonwoven fabric (inventive adsorption material-4) was obtained, in which the ligand was bis(3-aminopropyl)amine-acetyl-octylamide (the aromatic nucleus substituent was 2,5,9,13,16-pentaaza-3,15-dioxotetracosanyl). The picric acid adsorption capacity was 8.04 μmol / g.

[0070] <Preparation of the present invention adsorption material-6 (hexanoyl group type-1)> 4.9 g of coated nonwoven fabric-2 was immersed in a solution consisting of 1 g (3.3 mmol) of diethylenetriamine-hexanoylamide (prepared by reacting diethylenetriamine with an equimolar amount of hexanoyl chloride), 1 mL of 2N NaOH, and 40 mL of 50% aqueous dimethyl sulfoxide, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 4.8 g of nonwoven fabric (adsorbent material-6) in which the ligand was a diethyleneaminetriamine-hexanoylamide group (the aromatic nucleus substituent was a 2,5,9,13-tetraaza-3,14-dioxo-nonadecanoyl group) was obtained. The picric acid adsorption capacity was 30.2 μmol / g.

[0071] <Preparation of the present invention adsorption material-7 (hexanoyl group type-2)> 3.8 g of coated nonwoven fabric-3 was immersed in a solution consisting of 1 g (3.3 mmol) of diethylenetriamine-hexanoylamide, 1 mL of 2N NaOH, and 40 mL of 50% dimethyl sulfoxide, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 3.8 g of nonwoven fabric (inventive adsorption material-7) was obtained, in which the ligand was a diethylenetriamine-hexanoylamide group (the aromatic nucleus substituent was a 2,5,9,13-tetraaza-3,14-dioxo-nonadecanoyl group). The picric acid adsorption capacity was 5.2 μmol / g.

[0072] <Preparation of the present invention adsorption material-8 (hexanoyl group type-3)> 4.4 g of coated nonwoven fabric-4 was immersed in a solution consisting of 1 g (3.3 mmol) of diethylenetriamine-hexanoylamide, 1 mL of 2N NaOH, and 40 mL of 50% dimethyl sulfoxide water, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 4.4 g of nonwoven fabric (adsorbent material-8) in which the ligand was diethylenetriamine-hexanoylamide groups (the aromatic nucleus substituent was 2,5,9,13-tetraaza-3,14-dioxo-nonadecanoyl groups) was obtained. The picric acid adsorption capacity was 2.4 μmol / g.

[0073] <Preparation of the present invention adsorption material-9 (hexanoyl group type-4)> 2.9 g of coated nonwoven fabric-3 was immersed in a solution consisting of 1 g (3.3 mmol) of bis(3-aminopropyl)ethylenediamine-hexanoylamide (prepared by reacting an equimolar amount of hexanoyl chloride with bis(3-aminopropyl)ethylenediamine), 1 mL of 2N NaOH, and 40 mL of 70% ethanol, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 2.9 g of nonwoven fabric (adsorbent material-9) in which the ligand was bis(3-aminopropyl)ethylenediamine-hexanoylamide (2,5,9,13-tetraaza-3,14-dioxo-nonadecanoyl group as the aromatic nucleus substituent) was obtained. The picric acid adsorption capacity was 11.5 μmol / g.

[0074] <Preparation of the present invention adsorption material-10 (isobutyryl group type-1)> 3.6 g of coated nonwoven fabric-3 was immersed in a solution consisting of 1 g (5.7 mmol) of diethylenetriamine-isobutyrylamide (prepared by reacting diethylenetriamine with an equimolar amount of isobutyryl chloride), 1 mL of 2N NaOH, and 40 mL of 70% ethanol, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 3.6 g of nonwoven fabric (adsorbent material-10 of the present invention) was obtained, in which the ligand was a diethylenetriamine-isobutyrylamide group (the aromatic nucleus substituent was a 2,5,8,11-tetraaza-3,12-dioxo-13-methyl-tetradecanoyl group). The picric acid adsorption capacity was 4.7 μmol / g.

[0075] <Preparation of comparative adsorption material-1 (leucine base type)> 2.0 g (10 mmol) of chloroacetyl-L-leucine was added to a solution consisting of 2 mL (20 mmol) of diethylenetriamine, 200 mL of water, and 200 mL of dimethyl sulfoxide. The mixture was stirred at room temperature for 3 hours, after which 20 g of coated nonwoven fabric-2 was immersed and shaken in a 40°C water bath for 2 hours. The nonwoven fabric was washed with water, extracted three times with hot water at 60°C, and then dried to obtain 20 g of nonwoven fabric (comparative adsorption material-1) whose ligands were a mixture of acetyl-L-leucine and diethylenetriamine in a molar ratio of 1:2. The picric acid adsorption capacity was 159 μmol / g.

[0076] <Preparation of comparative adsorption material-2 (decanoyl group type)> 3.0 g of coated nonwoven fabric-3 was immersed in a solution consisting of 1 g (3.0 mmol) of bis(aminopropyl)ethylenediamine-decanoylamide (prepared by reacting an equimolar amount of decanoyl chloride with bis(aminopropyl)ethylenediamine), 1 mL of 2N NaOH, and 40 mL of ethanol, and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with 70% ethanol and then with water. After drying, 2.9 g of nonwoven fabric (comparative adsorption material-2) was obtained, in which the ligand was a bis(aminopropyl)ethylenediamine-decanoylamide group (the aromatic nucleus substituent was a 2,5,9,12,16-pentaaza-3,17-dioxohexacosane group). The picric acid adsorption capacity was 16.1 μmol / g.

[0077] <Preparation of comparative adsorption material-3 (diethylenetriamine type)> 3.0 g of coated nonwoven fabric-2 was immersed in a solution consisting of 1 g (3.0 mmol) of diethylenetriamine and 40 mL of dimethyl sulfoxide and heated in a 50°C water bath for 4 hours. The nonwoven fabric was washed with water and then dried to obtain 3.0 g of nonwoven fabric (comparative adsorption material-3) in which the ligand was a diethylenetriamino group (the aromatic nucleus substituent was a 10-amino-2,5,8-triazo-3-oxodecane group). The picric acid adsorption capacity was 108 μmol / g.

[0078] <Preparation of comparative adsorption material-4 (bisaminopropylamine type)> 3.0 g of coated nonwoven fabric-3 was immersed in a solution consisting of 1 g (3.0 mmol) of bis(3-aminopropyl)amine and 40 mL of 50% aqueous dimethyl sulfoxide solution and heated in a 50°C water bath for 4 hours. After rinsing with water, the nonwoven fabric was dried to obtain 3.0 g of nonwoven fabric (comparative adsorption material-4) in which the ligand was a bis(3-aminopropyl)amino group (the aromatic nucleus substituent was a 12-amino-2,5,9-triazo-3-oxododecano group). The picric acid adsorption capacity was 3.8 μmol / g.

[0079] <Preparation of comparative collection material-5 (polymyxin B type)> 3.0 g of coated nonwoven fabric-3 was immersed in 40 mL of an aqueous solution containing 0.2 g of polymyxin B and 0.5 mL of 2N NaOH and heated in a water bath at 50°C for 4 hours. The nonwoven fabric was washed with water and then dried to obtain 3.0 g of a nonwoven fabric (comparative adsorption material-5) whose ligand was polymyxin B. The picric acid adsorption capacity was 2.1 μmol / g.

[0080] [Test Example 1] (In vitro LAP cell adsorption evaluation) 0.1 g of collection material was added to 2 mL of blood collected from normal Donryu rats or WKAH / Hkm rats, and the material was shaken (120 rpm) at 37°C for 20 minutes. The LAP cell adsorption rate and the LAP-positive cell selection rate of the cell collection material were determined. The platelet retention rate, which indicates the suitability of the material for extracorporeal circulation, was also determined. + The results for T cells are shown in Table 1. + The results for T cells are shown in Table 2, and the results for B cells are shown in Table 3. Note that blood drawn from Donryu rats is abbreviated as D, blood drawn from WKAH / Hkm rats as K, and the numbers indicate the rat numbers.

[0081] [Table 1]

[0082] [Table 2]

[0083] [Table 3]

[0084] [Test Example 2] (Sepsis Treatment Experiment) Septic rats were anesthetized and administered LPS. Two hours later, extracorporeal circulation therapy was performed for 30 minutes using a column containing the present invention's adsorption material-1, a column containing the present invention's adsorption material-6, or an empty column containing no adsorption material, and the survival rates were compared. In addition, 0.5 mL of arterial blood was collected before and after extracorporeal circulation, and CD4 + The LAP positivity rate of T cells was determined. The results are shown in Table 4. All three rats treated with columns packed with inventive capture material-1 and inventive capture material-6 survived, but all three rats treated with empty columns and all three untreated rats died by the next day.

[0085] [Table 4]

Claims

1. An immunosuppressive cell capture material for selectively capturing immunosuppressive cells, comprising a molded body having on its surface an aromatic polymer having a substituent represented by the following general formula (1) in the aromatic residue of its main chain. φ-RXYZ (1) [wherein φ represents an aromatic nucleus in the main chain, R is -CH 2 -, -CH 2 NHCOCH 2 - or -CH 2 NHCOCH 2 CH 2 -, and X is -NH(CH 2 ) n -A-(CH 2 ) m NH- wherein n and m are the same or different and represent 2 or 3; A is -NH- or -NH-(CH 2 CH 2 ) p -NH-, p represents an integer between 2 and 4, Y is a carbonyl group, -CH 2 CONH- or -CH 2 CH 2 represents a CONH-group, Z represents a linear or branched alkyl group having 3 to 8 carbon atoms.

2. 2. The adsorbent according to claim 1, wherein the substituents represented by the general formula (1) are bonded at a frequency of 1 to 50 per 100 aromatic nuclei.

3. wherein X is a diethylenetriamine residue, a bis(3-aminopropyl)amine residue, or 1, The adsorption material according to claim 1 or 2, which is a 5,8,12-tetraazadodecane residue.

4. The capture agent according to any one of claims 1 to 3, wherein Z is an isoamyl group or a pentyl group. Collection of wood.

5. The collection material according to any one of claims 1 to 4, wherein the aromatic polymer is polysulfone, polyetherimide, polyimide, or a derivative thereof.

6. The collection material according to any one of claims 1 to 5, which is a molded product obtained by applying the aromatic polymer to fibers.

7. The immunosuppressive cells have latency-associated proteins on their cell surface. The collection material according to any one of claims 1 to 6, which is a lymphocyte.

8. A column for capturing immunosuppressive cells, packed with the capturing material according to any one of claims 1 to 7.

9. The column according to claim 8, which is for extracorporeal circulation.

10. The column of claim 8, which is for cell therapy.

11. The column according to any one of claims 8 to 10, which is used for treating an infection, treating an opportunistic infection, treating burns, preventing cancer recurrence after cancer removal surgery, treating cancer, or preventing sepsis.

Images