Biological component treatment material

Abstract

The purpose of the present invention is to provide a biological component treatment material that is configured to suppress a reduction in biological component treatment capability after autoclave sterilization. The present invention provides a biological component treatment material comprising a water-insoluble material containing: a first ligand having amino groups and amide groups; a second ligand having amide groups and hydrocarbon groups optionally substituted by hydroxy groups; and a substrate.

Description

Biocomponent Processing Materials

[0001] The present invention relates to a biological component processing material.

[0002] In recent years, technologies related to blood purification therapy have been developed for the treatment of fluids containing biological components, particularly for the purpose of treating inflammatory diseases, immunosuppression before and after transplantation, and suppressing side effects of blood products such as fever and infection, by adsorption and removal of blood components such as cytokines and leukocytes. Cytokines, which are biomarkers for inflammatory diseases, are preferred targets for adsorption, and technologies have been developed for the adsorption and removal of interleukin-6 (hereinafter referred to as IL-6) or interleukin-8 (hereinafter referred to as IL-8), which are types of cytokines.

[0003] Materials containing a ligand having an amide group and an amino group in the base material have excellent interaction with biological cells such as white blood cells and cytokines, and biological component treatment materials using such materials have been developed.

[0004] For example, Patent Document 1 discloses a biological component treatment material that is a water-insoluble material to which a ligand having an amide group and an amino group is bound, and that has improved blood purification performance by adjusting the amide group content and the amino group content within a certain range.

[0005] Patent Document 2 discloses a material for removing activated leukocyte-activated platelet complexes, which comprises a water-insoluble carrier containing a ligand and a substrate, the ligand having a hydrocarbon group and an amide group.

[0006] Patent No. 6589993 Patent No. 7331368

[0007] Columns containing biocomponent processing materials (hereinafter referred to as biocomponent processing columns) are primarily used for liquid processing, and are typically filled with a liquid (hereinafter referred to as filler liquid) to prevent air from being introduced during liquid circulation, thereby eliminating air. Since bacterial growth in the filler liquid can have a detrimental effect on biocomponent processing, biocomponent processing columns are sterilized. Methods for sterilizing liquid-containing materials include gamma ray sterilization and autoclaved steam sterilization. However, autoclaved steam sterilization is preferred because gamma ray sterilization requires specialized equipment for handling radioactive materials.

[0008] However, the present inventors have newly discovered that conventional biological component processing materials containing ligands having amino and amide groups have a problem in that their biological component processing ability, particularly their blood component adsorption ability, is reduced when subjected to high-pressure steam sterilization. If performance is reduced by high-pressure steam sterilization, the material will no longer function as a biological component processing column.

[0009] Therefore, an object of the present invention is to provide a biological component processing material that is prevented from decreasing in biological component processing ability due to high-pressure steam sterilization.

[0010] The present inventors have conducted extensive research to solve the above problems, and as a result have found the following inventions (1) to (4): (1) A biological component processing material comprising a water-insoluble material including a first ligand having an amino group and an amide group, a second ligand having a hydrocarbon group and an amide group, which may be substituted with a hydroxyl group, and a substrate, wherein the first ligand is bound to the substrate via a structure represented by the following general formula (I), and the second ligand is bound to the substrate via a structure represented by the following general formula (IIa) or (IIb). [In the formula, the wavy line represents the bonding position to the substrate, and X is a chemical structure that contains an amino group as a partial structure and may also contain a phenyl group.] [In the formula, the wavy line represents the bonding position to the substrate, A is an alkyl group or phenyl group having 1 to 6 carbon atoms which may be substituted with a hydroxy group, and n is an integer of 1 to 6.] (2) The biological component processing material according to (1), wherein the content of the second ligand is 0.3 to 5.0 mmol per gram of dry weight of the water-insoluble material. (3) The biological component processing material according to (1) or (2), wherein the content of amino groups in the first ligand is 1.0 to 4.0 mmol per gram of dry weight of the water-insoluble material. (4) A biological component processing column comprising the biological component processing material according to any one of (1) to (3).

[0011] The biological component processing material of the present invention can exhibit high biological component processing performance even when subjected to high-pressure steam sterilization.

[0012] The present invention will be described in detail below.

[0013] The term "substrate" refers to a material that can immobilize a first and second ligands by chemical modification and that is water-insoluble after immobilization. The substrate is, for example, a polymeric material containing, in its structure, repeating functional groups that are reactive with carbocations, such as aromatic rings or hydroxyl groups. Specific examples include synthetic polymeric materials such as poly(aromatic vinyl compounds) (e.g., polystyrene, polydivinylbenzene), polyesters (e.g., polyethylene terephthalate, polybutylene terephthalate), polysulfone, and polyvinyl alcohol, as well as natural polymeric materials such as cellulose, collagen, chitin, chitosan, and dextran. The substrate may also be a derivative of the above synthetic or natural polymeric materials to which alkyl groups, halogen groups, halogenated alkyl groups, acetal groups, ether groups, or the like have been added. Examples of polystyrene derivatives include poly(p-chloromethylstyrene), poly(α-methylstyrene), poly(β-methylstyrene), poly(p-tert-butoxystyrene), poly(p-acetoxystyrene), and poly(p-(1-ethoxyethoxy)styrene).

[0014] There are no particular restrictions on the composition of the polymer material that constitutes the substrate, but it may be a homopolymer or a copolymer of the above polymer materials, or a physical blend of multiple polymer materials.

[0015] Particularly for blood component treatment, the substrate is preferably a material that does not have hydroxyl groups, such as poly(aromatic vinyl compounds) (e.g., polystyrene, polydivinylbenzene), polyester (e.g., polyethylene terephthalate, polybutylene terephthalate), or polysulfone, or a derivative thereof, with polystyrene, polysulfone, or polydivinylbenzene, or a derivative thereof, being more preferred. Among these, polystyrene, polydivinylbenzene, or a derivative thereof is even more preferred because they have a large number of aromatic rings per unit weight and are easy to introduce ligands having amide groups and amino groups into.

[0016] The polymeric material used for the substrate may contain a crosslinked structure to maintain water insolubility even after ligand immobilization. While there is no limitation on the polymeric material containing a crosslinked structure, for example, a polymeric material in which a crosslinked structure has been introduced by copolymerizing a bifunctional monomer such as divinylbenzene, or a polymeric material in which a crosslinked structure has been introduced by reacting a crosslinking agent with a functional group such as an aromatic ring or a hydroxyl group in the polymeric material is preferred. From the perspective of ease of procurement, a polymeric material in which a crosslinked structure has been introduced by reacting a crosslinking agent with a functional group such as an aromatic ring or a hydroxyl group in the polymeric material is more preferred. As the crosslinking agent, formaldehyde or a substituted benzaldehyde (e.g., benzaldehyde, p-diethylaminobenzaldehyde (DEAP), p-dimethylaminobenzaldehyde (DMAP), or p-isopropylbenzaldehyde (IPP)) is preferred, with formaldehyde being more preferred.

[0017] The term "water-insoluble material" refers to a material that is insoluble in water, and the water-insoluble material of the present invention includes a substrate to which a first ligand and a second ligand are bound. Here, "water-insoluble" means that the change in dry weight of the material before and after placing it in water is 1% or less. This change in dry weight is determined by immersing the material in water at 37°C in an amount 9 times its dry weight, mixing by inversion for 1 hour, then removing the material with tweezers or the like, and vacuum-drying the remaining water at 50°C or less, and measuring the dry weight of the remaining solids relative to the dry weight of the material before immersion in water. If the material is not water-insoluble, there is a risk of excessive elution during actual use, which is undesirable from a safety standpoint.

[0018] "Dry weight" refers to the weight of a solid in a dry state. A dry solid refers to a solid in which the amount of liquid component contained in the solid is 1% by weight or less. After measuring the weight of a solid, the solid is dried by heating at 80°C at atmospheric pressure for 24 hours, and when the weight loss of the remaining solid is 1% by weight or less of the weight before drying, the solid is considered to be a dry solid. In the present invention, unless otherwise specified, all weights refer to dry weight.

[0019] The term "ligand" refers to a chemical structure contained in a water-insoluble material and exhibiting a specific function. The water-insoluble material of the present invention includes a first ligand having an amino group and an amide group, and a second ligand having an amide group and a hydrocarbon group optionally substituted with a hydroxy group, the first ligand being bound to the substrate via a structure represented by the following general formula (I), and the second ligand being bound to the substrate via a structure represented by the following general formula (IIa) or (IIb):

[0020] The term "first ligand" refers to a ligand having an amino group and an amide group and exhibiting the ability to process a biological component, and has a structure represented by the following general formula (I). [In the formula, the wavy line represents the bonding position to the substrate, and X is a chemical structure that contains an amino group as a partial structure and may also contain a phenyl group.]

[0021] Furthermore, the first ligand may have a structure represented by the following general formula (III), i.e., a structure having an amide group, an amino group, and a phenyl group, and this structure is preferable because it can further inhibit platelet adhesion. [In the formula, Y is an amino group, Z is a linker, R is a hydrogen atom or a halogen atom, and the wavy line represents the bonding position with the substrate.]

[0022] Since the electron density of the amino group is controlled by the amide group, the amino group and the amide group are preferably covalently bonded via an alkylene group. The alkylene group is preferably a saturated hydrocarbon structure having 5 or fewer carbon atoms, such as a pentylene group, a butylene group, a propylene group, an ethylene group, or a methylene group, with a methylene group being more preferred. Furthermore, in the ligand having the amide group and the amino group, the amide group side is bonded to the substrate. In the first ligand, the functional groups contained other than the amide group and the amino group are not particularly limited, but may include, for example, a phenyl group (the phenyl group may have a substituent such as a halogen atom, a halogenated alkyl group, or a linear alkyl group having 1 to 5 carbon atoms). In this case, the phenyl group is preferably bonded to the amino group via a linker, as described below.

[0023] The term "amino group" refers to a chemical structure derived from an amine compound, and examples thereof include amino groups derived from primary amines such as methylamine, ethylamine, propylamine, butylamine, pentylamine, hexylamine, heptylamine, octylamine, and dodecylamine; amino groups derived from secondary amines such as methylhexylamine, diethylamine, and dimethylamine; amino groups derived from amines with unsaturated alkyl chains such as allylamine; and amino groups derived from tertiary amines such as trimethylamine, triethylamine, dimethylethylamine, phenyldimethylamine, and dimethylhexylamine. Examples of the amino group include a conventional amino group, an amino group derived from an amine having an aromatic ring, such as 1-(3-aminopropyl)imidazole, pyridine-2-amine, or aniline, and an amino group derived from a compound in which two or more amino groups are bonded via an alkyl chain, an aromatic compound, a heterocyclic compound, a homocyclic compound, or the like (hereinafter referred to as a "polyamine"), such as tris(2-aminoethyl)amine, ethylenediamine, diethylenetriamine, triethylenetetramine, tetraethylenepentamine, dipropylenetriamine, polyethyleneimine, or N-methyl-2,2'-diaminodiethylamine. The amino group contained in the first ligand is preferably an amino group derived from a polyamine, more preferably an amino group derived from ethylenediamine, diethylenetriamine, triethylenetetramine, tetraethylenepentamine, or polyethyleneimine, and even more preferably an amino group derived from tetraethylenepentamine or polyethyleneimine.

[0024] The number of carbon atoms per nitrogen atom of the amino group contained in the first ligand is preferably 18 or less, more preferably 14 or less, and even more preferably 8 or less, taking into consideration nucleophilicity and steric hindrance, which affect the reaction rate. The nitrogen atom of the amino group is preferably substituted with a hydrocarbon group. The hydrocarbon group may be a linear hydrocarbon group, a branched hydrocarbon group, a cyclic hydrocarbon group, or a hydrocarbon group that is a combination thereof. Examples of the hydrocarbon group include alkyl groups and alkylene groups. The alkyl group is preferably a linear alkyl group such as a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, a heptyl group, or an octyl group, and more preferably a methyl group, an ethyl group, or a propyl group. The alkylene group is preferably a linear alkylene group such as a methylene group, an ethylene group, a propylene group, a butylene group, a hexylene group, a heptylene group, or an octylene group, and more preferably a methylene group, an ethylene group, or a propylene group.

[0025] The term "phenyl group" refers to a chemical structure derived from unsubstituted benzene or a substituted benzene compound, and examples thereof include benzene, fluorobenzene, chlorobenzene, bromobenzene, 1,2-difluorobenzene, 1,2-dichlorobenzene, 1,2-bromobenzene, 1,3-difluorobenzene, 1,3-dichlorobenzene, 1,3-dibromobenzene, 1,4-difluorobenzene, 1,4-dichlorobenzene, and 1,4-dibromobenzene. In order to control the charge of the amino group, a phenyl group derived from a halogenated benzene to which an electron-withdrawing group has been added is preferred, and among these, a chlorophenyl group derived from chlorobenzene is preferred. From the viewpoint of a resonance structure, the electron-withdrawing group is preferably added at the para position, and in particular, in the case of a phenyl group derived from chlorobenzene, a parachlorophenyl group in which the linker and the chlorine atom are substituted at the para position is preferred.

[0026] "Halogen atom" means a fluorine atom, chlorine atom, bromine atom or iodine atom.

[0027] The term "linker" refers to a chemical bond between the amino group and the phenyl group, and examples thereof include electrically neutral chemical bonds such as an amide bond, a urea bond, an ether bond, or an ester bond, with an amide bond or a urea bond being preferred.

[0028] The term "secondary ligand" refers to a ligand having a hydrocarbon group and an amide group, which may be substituted with a hydroxy group, and which does not inhibit the ability of the primary ligand to process biological components, and has a structure represented by the following general formula (IIa) or (IIb): [In the formula, the wavy line represents the bonding position to the substrate, A represents an alkyl group having 1 to 6 carbon atoms or a phenyl group which may be substituted with a hydroxy group, and n represents an integer of 1 to 6.]

[0029] If the structure of the second ligand is too long, its mobility may increase, and there is a concern that the interaction with the first ligand may result in a decrease in the ability to process biological components. Therefore, A is an alkyl group or phenyl group having 1 to 6 carbon atoms which may be substituted with a hydroxy group, preferably an alkyl group or phenyl group having 1 to 3 carbon atoms which may be substituted with a hydroxy group, more preferably a methyl group, a hydroxymethyl group, a phenol group, or a phenyl group, and even more preferably a methyl group or a hydroxymethyl group.

[0030] The term "amide group" refers to an amide bond contained in the first and second ligands.

[0031] There are no particular limitations on the content of amino groups in the first ligand per 1 g of dry weight of the water-insoluble material, but if the content is too small, the adsorption performance for blood components, among other biological component treatment capabilities, will not be exhibited, whereas if the content is too large, the ligand density will be too high, resulting in reduced mobility and failure to exhibit the adsorption performance for blood components. Therefore, the content is preferably 1.0 to 4.0 mmol, and more preferably 1.2 to 3.8 mmol.

[0032] There is no particular limitation on the content of amide groups in the first ligand per 1 g of the dry weight of the water-insoluble material, but if the content is too small, it becomes impossible to introduce amino groups, and if the content is too large, it becomes impossible to introduce the second ligand. Therefore, a content of 1.0 to 7.0 mmol is preferred.

[0033] There are no particular limitations on the content of phenyl groups in the first ligand per 1 g of the dry weight of the water-insoluble material; however, if the content is too small, the effect of inhibiting platelet adhesion will not be exhibited, and if the content is too large, the performance of removing humoral factors from blood will decrease; therefore, a content of more than 0 to 7.0 mmol is preferred.

[0034] There are no particular limitations on the content of the second ligand per 1 g of the dry weight of the water-insoluble material, but if the content is too small, the content of the second ligand on the surface of the water-insoluble material is likely to be uneven, resulting in a lack of production stability, while if the content is too large, the material is likely to deform and handleability is poor. Therefore, the content is preferably 0.3 to 5.0 mmol, and more preferably 0.4 to 3.0 mmol.

[0035] The term "biological component processing material" refers to a material for processing biological components. As long as the material has the ability to process biological components, there are no particular limitations on whether or not it has the ability to process other substances. The biological component processing material of the present invention is preferably used for adsorbing and removing blood components. The biological component processing material of the present invention may contain a water-insoluble material containing a substrate and a first and second ligands, and may also contain a reinforcing material mixed in to maintain the shape of the water-insoluble material and properly demonstrate its functionality. The reinforcing material may be mixed in at the stage of the substrate alone, before it becomes a water-insoluble material, or after the first and second ligands have been bound to the substrate.

[0036] A "reinforcing material" is a material that is intentionally included in a biological component processing material to maintain the shape of the water-insoluble material and allow liquids containing biological components to properly contact the water-insoluble material, and does not itself exhibit the main biological component processing performance.

[0037] The combination of the reinforcing material and the water-insoluble material is not particularly limited, and examples thereof include a combination in which the island component of a sea-island composite fiber is a reinforcing material and the sea component is a water-insoluble material, a combination in which one component of a polymer alloy fiber is a water-insoluble material and the other component is a reinforcing material, and a combination in which the nonwoven fabric of a nonwoven fabric-like material is a reinforcing material and the mixed cut fibers are a water-insoluble material.

[0038] The chemical structure of the reinforcing material is not particularly limited, and examples thereof include homopolymers containing only one monomer selected from the group consisting of ethylene glycol, butylene glycol, terephthalic acid, aromatic vinyl compounds (e.g., styrene, divinylbenzene), glucose, glucose triacetate, vinylpyrrolidone, vinyl alcohol, acrylonitrile, sodium methallylsulfonate, ethylene, propylene, ε-caprolactam, and methyl methacrylate; copolymers containing two or more monomers selected from the above group; and physical blends of the above homopolymers and copolymers. To avoid inhibiting adsorption by water-insoluble materials, the reinforcing material is preferably a polymer containing a compound that does not contain an aromatic ring and / or a hydroxyl group as a monomer. Specific examples include homopolymers containing only one monomer selected from the group consisting of vinylpyrrolidone, acrylonitrile, sodium methallylsulfonate, ethylene, propylene, ε-caprolactam, and methyl methacrylate; copolymers containing two or more monomers selected from the above group; and physical blends of the above homopolymers and copolymers. Among these, polymers containing ethylene and / or polypropylene as monomers (e.g., polyethylene, polypropylene) are preferred from the viewpoint of high chemical resistance and resistance to deterioration. Here, when two or more polymers are completely miscible or when only one of the polymers in the block copolymer contains the first ligand and the second ligand, the other polymer is not defined as a reinforcing material but is regarded as a substrate.

[0039] The shape of the biological component treatment material may be a fiber, flat membrane, hollow fiber membrane, or particle, with the fiber or particle shapes being preferred from the viewpoint of not forming retention areas upon contact with biological components. Furthermore, among the above-mentioned fiber shapes, thread bundles, yarns, nets, knitted fabrics, woven fabrics, etc., which are processed from the above-mentioned fibers are preferred, with the thread bundles, knitted fabrics, and woven fabrics being more preferred in terms of their large surface area and low flow resistance.

[0040] When the biological component processing material is in particulate form, the particle diameter may be any size, but from the viewpoint of increasing the contact area and preventing clogging, it is preferably 1 μm to 1 mm, more preferably 20 μm to 500 μm.

[0041] When the biological component processing material is in the form of fibers, the diameter of the single fiber may be any thickness, but from the viewpoint of improving the contact area and maintaining the strength of the material, it is preferably 3 to 200 μm, more preferably 5 to 50 μm, and even more preferably 10 to 40 μm.

[0042] The "single fiber diameter" means the average value of the measured values ​​obtained by randomly collecting 10 small fiber samples, taking photographs of each sample using a scanning electron microscope (SEM) or the like, and measuring the fiber diameter at 10 points in each photograph (100 points in total for all samples).

[0043] When the biological component processing material is in the form of fibers, the cross-sectional structure of the fibers may be, for example, a single thread made of one type of polymer, or a core-sheath, sea-island, or side-by-side composite fiber. From the viewpoint of maintaining the strength of the material during biological component processing, core-sheath composite fibers or sea-island composite fibers are preferred.

[0044] Biocomponent treatment materials in the form of knitted fabric, felt, or net can be produced by known methods using a fibrous substrate or biocomponent treatment material as a raw material. Examples of methods for producing felt include the wet method, carding method, air-laying method, spunbonding method, and melt-blowing method. Examples of methods for producing knitted fabrics and nets include the plain weave method and tubular knitting method. In particular, knitted fabrics produced by the tubular knitting method are preferred, as they have a large fill mass per unit volume and can be packed into blood purifiers.

[0045] When the cross-sectional structure of the fiber is an islands-in-sea type solid composite fiber, the constituent components are not particularly limited, but from the viewpoint of ensuring strength while exhibiting performance, it is preferable that the sea part is a water-insoluble material and the island parts are reinforcing materials. For example, an islands-in-sea type solid composite fiber in which the sea part is made of a water-insoluble material and the island parts are made of polyolefin can be mentioned, and a specific example is an islands-in-sea type solid composite fiber in which the sea part is a water-insoluble material containing polystyrene as a base material, a first ligand, and a second ligand, and the island parts are made of polypropylene.

[0046] The term "biological component" refers to a component contained in a living organism, and is not limited by its chemical or physical structure, but examples include fat and oil components, blood components, lymph components, viruses, bacteria, etc. There are no particular limitations on the biological components that can be treated with the biological component treatment material of the present invention, but the treatment targets are preferably biological components such as blood components, lymph components, viruses, and bacteria that have an electric charge in their structure that interacts with the first ligand and that are present primarily in liquid, with blood components being more preferred, and leukocyte components and cytokines being even more preferred, particularly when the treatment is aimed at treating inflammatory diseases.

[0047] "Blood components" refer to components that make up blood and are classified into liquid components in blood and cells in blood. There are no particular restrictions on the blood components that can be treated with the biological component treatment material of the present invention, but liquid components in blood are preferred.

[0048] "Cells in blood" means cells contained in blood, and examples thereof include white blood cell components such as granulocytes, monocytes, or lymphocytes, as well as red blood cells or platelets. When the purpose is to treat an inflammatory disease, the target of treatment is preferably white blood cell components, and among white blood cell components, preferred are monocytes and granulocytes (including activated granulocytes, activated monocytes, activated granulocyte-activated platelet complexes, or activated monocyte-activated platelet complexes), which release inflammatory cytokines and whose removal can calm inflammatory diseases.

[0049] "Activated granulocytes" and "activated monocytes" refer to granulocytes and monocytes, respectively, that release cytokines or reactive oxygen species in response to cytokines, lipopolysaccharide (LPS), etc. The degree of activation can be determined by measuring the amount of reactive oxygen species released by activated leukocytes or by measuring the expression of surface antigens by flow cytometry, etc.

[0050] The term "activated platelets" refers to platelets that release cytokines, active oxygen, etc. in response to cytokines, LPS, etc.

[0051] The terms "activated granulocyte-activated platelet complex" and "activated monocyte-activated platelet complex" refer to activated granulocytes or activated monocytes bound to activated platelets, which have the ability to phagocytose autologous tissue. In particular, in the treatment of patients with inflammatory diseases, it is considered important to remove activated granulocyte-activated platelet complexes, which are thought to be directly involved in the pathology.

[0052] "Liquid components in blood" refers to organic substances dissolved in blood. Specific examples include low molecular weight organic compounds such as urea or amino acids, proteins such as β2-microglobulin, cytokines, IgE or IgG, and polysaccharides such as LPS. Among these, proteins such as cytokines or polysaccharides such as LPS are preferred as treatment targets, and when the purpose is to treat inflammatory diseases, cytokines are more preferred as treatment targets.

[0053] The term "cytokine" refers to a group of proteins that are produced by various cells, including immunocompetent cells, in response to stimuli such as infection or trauma, and are released extracellularly to act, and examples thereof include interferon α, interferon β, interferon γ, interleukin 1 to interleukin 15, tumor necrosis factor α, tumor necrosis factor β, high mobility group box-1, erythropoietin, and monocyte chemotactic factor. Among these, interleukin 1β, IL-6, IL-8, high mobility group box-1, tumor necrosis factor β, etc. are preferred as targets of treatment, and in the treatment of inflammatory diseases, IL-6 or IL-8 is more preferred as targets of adsorption.

[0054] The term "treatment" refers to the contact of a specific substance with a water-insoluble material and the resulting change from its state before the contact, and examples of such treatment include inactivation, activation, and adsorption. From the viewpoint of treating inflammatory diseases, inactivation or adsorption is preferred, with adsorption being more preferred.

[0055] "Inactivation" means that a specific substance comes into contact with a water-insoluble material, changes from its liquid state, and loses its original function. For example, a protein loses its three-dimensional structure due to electrostatic or hydrophobic interaction with the surface of a water-insoluble material, and loses its ability to interact with other substances, or activated leukocytes or platelets lose their activity upon contact with the surface of a water-insoluble material, returning to an inactivated state.

[0056] "Activation" means that a specific substance comes into contact with a water-insoluble material and changes from a liquid state to a state in which a specific function is expressed. For example, a denatured protein restores its three-dimensional structure through electrostatic or hydrophobic interaction with the surface of a water-insoluble material, thereby expressing the function of interacting with other substances, or non-activated leukocytes or platelets become activated upon contact with the surface of a water-insoluble material.

[0057] "Adsorption" refers to a state in which a specific substance adheres to a water-insoluble material and does not easily peel off. Examples of the principle of adsorption include adhesion due to ionic interactions such as electrostatic interactions, van der Waals forces such as hydrophobic interactions or hydrogen bonds, and biological adhesion such as cell adhesion and phagocytosis by leukocytes.

[0058] The biological component processing material of the present invention is also preferably used as a biological component processing column. The use of the biological component processing column is not particularly limited, but is preferably used to remove unwanted biological components from a solution, and more preferably used to remove proteins (e.g., cytokines) from a solution. Specific examples include blood purification therapy, which removes pathogenic substances from blood (e.g., treatment by removing cytokines from the blood of patients with inflammatory diseases), and removal of unwanted biological components from solutions containing biological components (e.g., removal of cytokines from organ preservation solutions intended to maintain the preservation state of organs removed for transplantation, removal of cytokines during the production of cell preparations such as blood products, removal of biological components other than T cells during T cell extraction in CAR-T therapy, removal of cytokines and cell membranes from iPS cell-derived platelets, regeneration of dialysate and other liquids by treating unwanted proteins and urea in the dialysate, and removal of unwanted bacterial proteins during the production of biopharmaceuticals). Among these, the biological component processing column is preferably used in blood purification therapy, in which removal of inflammatory cytokines is effective.

[0059] When the biological component adsorption column of the present invention is used as a blood purifier for blood purification therapy, the column may be in any shape as long as it has a blood inlet and outlet, and examples of such a container include cylindrical containers and prismatic containers such as triangular, quadrangular, hexagonal, and octagonal prisms. Blood drawn from the body may be passed directly through the column, or it may be used in combination with other body fluid treatment methods or medical devices. Examples of other body fluid treatment methods and medical devices include plasma exchange, peritoneal dialysis, plasma separators, hemofilters, cardiopulmonary bypass, and ECMO.

[0060] The biological component processing column of the present invention is particularly suitable for use in blood purification therapy, particularly in the treatment of inflammatory diseases. When used for the treatment of inflammatory diseases, a preferred extracorporeal circulation method involves connecting a biological component processing column containing the biological component processing material to a patient via a blood circuit, passing a body fluid extracted from the patient through the biological component processing column, and returning the body fluid to the patient. From the viewpoint of suppressing further inflammation induced by blood components, continuous processing is preferred, with a duration of 4 hours or more being more preferred, and a duration of 24 hours or more being even more preferred.

[0061] "Inflammatory disease" refers to all diseases that induce an inflammatory response in the body, and includes, for example, systemic lupus erythematosus, malignant rheumatoid arthritis, multiple sclerosis, ulcerative colitis, Crohn's disease, drug-induced hepatitis, alcoholic hepatitis, hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E, sepsis (e.g., gram-negative bacterial sepsis, gram-positive bacterial sepsis, culture-negative sepsis, or fungal sepsis), influenza, acute respiratory distress syndrome (ARDS; also referred to as acute respiratory distress syndrome or acute respiratory distress syndrome), acute lung injury (ALI), pancreatitis, idiopathic interstitial pneumonia (IDP), and the like. These include idiopathic pulmonary fibrosis (IPF), transfusion of blood products, organ transplantation, reperfusion injury after organ transplantation, cholecystitis, cholangitis, and neonatal blood type incompatibility.

[0062] Among inflammatory diseases, drug-induced hepatitis, alcoholic hepatitis, hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E, sepsis (e.g., gram-negative bacterial sepsis, gram-positive bacterial sepsis, culture-negative sepsis, or fungal sepsis), influenza, acute respiratory distress syndrome, acute lung injury, pancreatitis, or idiopathic interstitial pneumonia are preferred targets for treatment because causative substances are released into the blood, and blood purification is particularly expected to have a therapeutic effect. Preferred uses of the biological component processing column of the present invention include the treatment of the above-mentioned inflammatory diseases, and more preferred uses include the treatment of sepsis (e.g., gram-negative bacterial sepsis, gram-positive bacterial sepsis, culture-negative sepsis, or fungal sepsis), influenza, acute respiratory distress syndrome, acute lung injury, or idiopathic interstitial pneumonia, which are difficult to treat with drugs alone and are considered to involve both cytokines and activated leukocytes and activated platelets.

[0063] The biological component processing material of the present invention can be produced, for example, by the following method, but is not limited to this method.

[0064] Commercially available substrates can be used as they are. When desired molding into a fiber, flat membrane, hollow fiber membrane, or particle shape is desired, commercially available resins can be purchased, heated and melted, and extruded from a spinneret, or dissolved in a good solvent and then extruded from a spinneret into a poor solvent or into the atmosphere, thereby molding the substrate into the desired shape. Alternatively, particle-shaped substrates can be produced by dissolving a polymer material to be used as the substrate in a good solvent, dripping the polymer solution into a poor solvent, and then removing the solvent to cause the polymer to aggregate, or by precipitation simultaneously with polymerization through suspension polymerization. When a reinforcing material is mixed into the substrate, it can be mixed by, for example, mixing and molding during processing of the substrate, or by physically mixing in the material by needle punching after processing.

[0065] A substrate containing an amide group of a first ligand and a second ligand can be produced by adding the substrate, or the substrate and a reinforcing material, to a solution in a nonpolar solvent in which a catalyst, chloroacetamide having a haloalkyl group and a hydroxy group as a compound from which the amide group of the first ligand is derived, and acetamide having a hydrocarbon group and a hydroxy group as a compound from which the second ligand is derived, are dissolved, and then reacting the solution.

[0066] The catalyst used may be, for example, sulfuric acid, hydrochloric acid, nitric acid, aluminum (III) chloride, or iron (III) chloride, with sulfuric acid or iron (III) chloride being preferred.

[0067] As a chloroacetamide having a haloalkyl group and a hydroxy group as a compound from which the amide group of the first ligand is derived, for example, N-hydroxymethyl-2-chloroacetamide (hereinafter, referred to as NMCA) can be used.

[0068] As the compound from which the second ligand is derived, an acetamide having a hydrocarbon group and a hydroxy group can be used, for example, N-(hydroxymethyl)acetamide (hereinafter, NMA), N-(hydroxymethyl)benzamide (hereinafter, NMBA), or N-[4-(hydroxymethyl)phenyl]acetamide (hereinafter, NMPA).

[0069] Alternatively, acetamide having a hydrocarbon group and a hydroxy group as the compound from which the second ligand is derived can be produced by dissolving a primary amide compound having a hydrocarbon group which may be substituted with a hydroxy group, paraformaldehyde, and a catalyst in a solvent to form acetamide having a hydrocarbon group and a hydroxy group as the compound from which the second ligand is derived in the system, and then mixing with the compound from which the amide group of the first ligand is derived, and adding the substrate, or the substrate and reinforcing material, to this and allowing it to react, thereby producing a substrate containing the amide group of the first ligand and the second ligand.

[0070] As a primary amide compound having a hydrocarbon group which may be substituted with a hydroxy group, for example, 2-hydroxyacetamide can be used.

[0071] Examples of the solvent include nitrobenzene, nitropropane, chlorobenzene, toluene, and xylene, with nitrobenzene or nitropropane being preferred.

[0072] The catalyst may be, for example, sulfuric acid, hydrochloric acid, nitric acid, aluminum(III) chloride, or iron(III) chloride, with sulfuric acid or iron(III) chloride being preferred.

[0073] When a crosslinked structure is to be introduced into the substrate, a crosslinking agent may be added at the same time. Examples of the crosslinking agent that can be used include paraformaldehyde (hereinafter referred to as PFA), acetaldehyde, and benzaldehyde.

[0074] Examples of non-polar solvents include nitrobenzene, nitropropane, chlorobenzene, toluene, and xylene, with nitrobenzene or nitropropane being preferred.

[0075] The catalyst, chloroacetamide having a haloalkyl group and a hydroxy group as the compound from which the amide group of the first ligand is derived, acetamide having a hydrocarbon group and a hydroxy group as the compound from which the second ligand is derived, the crosslinking agent, and the nonpolar solvent may be commercially available.

[0076] In the reaction between the compound from which the amide group of the first ligand is derived, the compound from which the second ligand is derived, and the substrate, the catalyst concentration in the reaction solution is preferably 5 to 80 wt %, more preferably 30 to 70 wt %. The reaction temperature is preferably 0 to 90°C, more preferably 5 to 40°C. The reaction time is preferably 1 minute to 120 hours, more preferably 5 minutes to 24 hours.

[0077] A water-insoluble material comprising a first ligand, a second ligand, and a substrate can be produced, for example, by adding a substrate comprising the amide group of the first ligand and the second ligand to a polar solvent in which a compound having an amino group (hereinafter also referred to as an amine compound) is dissolved, and then reacting the substrate. Furthermore, the amine introduced as the first ligand can be further modified by subsequently adding a compound reactive with the amino group (e.g., chlorophenyl isocyanate) to the polar solvent and allowing the reaction to occur. The timing of modifying the amine compound is not particularly limited. The modification reaction may be carried out immediately after the introduction of the amine compound, as described above, or the amine compound may be reacted with a compound reactive with an amino group prior to reacting the substrate comprising the amide group of the first ligand and the second ligand. In the latter case, the modified compound having an amino group obtained by reacting the amine compound with the compound reactive with an amino group can be introduced as the first ligand into the substrate comprising the amide group of the first ligand and the second ligand.

[0078] As the amine compound, for example, polyethyleneimine, hexaethylenetetramine, tetraethylenepentamine, triethylenetetramine, diethylenetriamine, or ethylenediamine can be used.

[0079] As a compound reactive with an amino group, for example, chlorophenyl isocyanate can be used.

[0080] Examples of polar solvents include N,N-dimethylformamide, diethyl ether, dioxane, tetrahydrofuran, and dimethyl sulfoxide, with N,N-dimethylformamide and dimethyl sulfoxide being preferred.

[0081] In the reaction between the substrate containing the amide group of the first ligand and the second ligand and the compound having an amino group, the reaction temperature is preferably 10 to 90° C., more preferably 30 to 60° C. The reaction time is preferably 1 minute to 120 hours, more preferably 5 minutes to 24 hours.

[0082] The weight percentage of the water-insoluble material in the biocomponent processing material can be calculated by removing the reinforcing material from a specified amount of biocomponent processing material, measuring the weight of the remaining water-insoluble material, and dividing the weight by the weight of the biocomponent processing material before removal. Removal of the reinforcing material from the biocomponent processing material can be achieved by physical separation if no welding is observed visually or by morphological observation using an SEM. In the case of polyolefins to which the reinforcing material is welded, removal can be achieved by immersing the biocomponent processing material in xylene, refluxing it at its boiling point, dissolving it, and removing the residue.

[0083] The dry weight is determined by measuring the weight of the solid, then heating and drying it at 80°C and atmospheric pressure for 24 hours, and if the weight loss of the remaining solid is 1% by weight or less of the weight before drying, the solid is considered to be in a dry state, and the weight at that time can be used as the dry weight. If the weight loss exceeds 1% by weight, the solid is again heated and dried at 80°C and atmospheric pressure for 24 hours, and this process can be repeated until the weight loss falls below 1% by weight, thereby achieving a dry state.

[0084] When measuring the content of amino groups or phenyl groups in the first ligand or the content of the second ligand per gram of dry weight of the water-insoluble material, the content of amino groups or phenyl groups in the first ligand or the content of the second ligand can be calculated by measuring the content of amino groups or phenyl groups in the first ligand or the content of the second ligand in the biological component treatment material after drying, and dividing the obtained value by the product of the weight of the biological component treatment material and the weight proportion of the water-insoluble material in the biological component treatment material (the weight of the water-insoluble material in the biological component treatment material).

[0085] The content of amino groups in the first ligand per gram of dry weight of the water-insoluble material contained in the biological component treatment material can be determined, for example, by measuring the dry weight of the biological component treatment material after drying, ion-exchanging the amino groups in the biological component treatment material with hydrochloric acid, back-titrating with an aqueous sodium hydroxide solution, and dividing the obtained value by the product of the weight of the biological component treatment material and the weight proportion of the water-insoluble material in the biological component treatment material.

[0086] The content of phenyl groups in the first ligand per gram of dry weight of the water-insoluble material contained in the biological component treatment material can be determined, for example, by measuring the dry weight of the biological component treatment material after drying, heating the biological component treatment material in hydrochloric acid to hydrolyze the material, and then extracting aromatic compounds derived from phenyl groups contained in the solution. １ The aromatic compound concentration in the solution can be determined by measuring with H-NMR and creating a calibration curve using an internal standard to calculate the concentration, and then dividing the calculated concentration by the product of the weight of the biological component treatment material and the weight proportion of the water-insoluble material in the biological component treatment material.

[0087] The content of the second ligand per gram of dry weight of the water-insoluble material contained in the biological component treatment material can be determined, for example, by measuring the dry weight of the biological component treatment material, hydrolyzing the biological component treatment material by heating it in hydrochloric acid, and determining the amount of carboxylic acid derived from the second ligand contained in the solution. １ The carboxylic acid concentration in the solution can be determined by measuring with H-NMR and creating a calibration curve using an internal standard to calculate the carboxylic acid concentration in the solution, and then dividing the calculated concentration by the product of the weight of the biological component treatment material and the weight proportion of the water-insoluble material in the biological component treatment material.

[0088] One method for evaluating the blood component adsorption capacity of a biological component treatment material is to impregnate the biological component treatment material with fetal bovine serum (FBS) in which cytokines have been dissolved, measure the amount of decrease in cytokine concentration in the FBS after impregnation, and calculate the cytokine adsorption rate.

[0089] Cytokines are a type of protein released into the blood, and the greater the reduction in cytokine concentration due to impregnation with a biological component processing material, the higher the blood component adsorption ability of the biological component processing material can be determined to be.

[0090] The adsorption of IL-6 or IL-8 to the biocomponent processing material is thought to be a Langmuir-type equilibrium reaction that coats the surface with a single layer, and therefore is thought to reach adsorption equilibrium after approximately 4 hours of adsorption treatment, regardless of the concentration of the substance to be adsorbed. Therefore, the IL-6 or IL-8 adsorption rate of the biocomponent processing material is preferably 100% after 4 hours of adsorption treatment, and since this rate is time-dependent, the biocomponent processing material can be considered to have sufficient IL-6 or IL-8 adsorption capacity if it is 50% or higher after 2 hours of adsorption treatment.

[0091] The rate of performance decline of a biological component processing material after autoclave sterilization can be calculated by calculating the IL-6 or IL-8 adsorption rate of the biological component processing material before and after autoclave sterilization, and then calculating the ratio of the decline in adsorption rate before and after autoclave sterilization. Performance decline due to autoclave sterilization is undesirable because it prevents the biological component processing material from achieving the target performance it was designed for. Considering variations in biocomponent adsorption, a decline in performance of 20% or more before and after autoclave sterilization can be considered to indicate a decline in performance.

[0092] The pH of the filling liquid for the biological component processing material after high-pressure steam sterilization can be determined by performing high-pressure steam sterilization while the biological component processing material is immersed in the filling liquid, removing the biological component processing material, and measuring the pH of the filling liquid.

[0093] If the filler solution becomes acidic or basic as a result of autoclaving a biological component processing column, there is a risk of adverse effects on the biological components being handled, and so it is necessary to pre-wash the column until it becomes neutral. Therefore, the further the pH of the filler solution after autoclaving deviates from 7, the longer the washing time becomes, making it difficult to handle. If the filler solution becomes more acidic than pH 4 or more basic than pH 11, it is necessary to remove 99.9% of the filler solution, or it can be considered difficult to handle because it cannot be neutralized without neutralization.

[0094] The biocomponent treatment material of the present invention will be described in detail below using examples, but the present invention is not limited to these examples. In the examples, wt% means weight %, M means mol / L, and mM means mmol / L. Unless otherwise specified, the weights of the substrate, water-insoluble material, and biocomponent treatment material are dry weights. Total fineness refers to the weight (grams) per 10,000 m of fiber and is expressed as dtex. pH measurements in acid-base titration were performed by immersing the electrode of a HORIBA benchtop pH meter F-74BW (includes a Standard ToupH electrode 9615S-10D) in a solution at 25°C. Prior to pH measurement, calibration was performed using a pH standard solution set [phthalate standard solution (pH 4.01, Horiba, Ltd.), neutral phosphate standard solution (pH 6.86, Horiba, Ltd.), and borate standard solution (pH 9.18, Horiba, Ltd.)]. For the NMR measurement, a JNM-EZ400R (manufactured by JEOL Ltd.) was used. １ The H-NMR measurement conditions were an accumulation of 32 times and a relaxation time of 5 seconds, and the subsequent NMR measurements were performed under the same conditions unless otherwise specified.

[0095] <Preparation of biological component processing material>

[0096] (Preparation of Substrate A) Polystyrene was used as the sea component and polypropylene as the island component, and these were separately melted and weighed, and the melts were fed into a spin pack equipped with a sea-island composite spinneret having 704 distribution holes for the island components per nozzle to form a sea-island composite stream, which was then melt-extruded. The island ratio was controlled to 50 wt %, and islands-in-sea type solid composite fibers with a single fiber size of 3.0 dtex (fiber diameter 20 μm) were obtained. The density adjustment scale of a cylindrical knitting machine (model name: circular knitting machine MR-1, Maruzen Sangyo Co., Ltd.) was adjusted to obtain a basis weight of 39 g / m ２ , bulk density is 0.22 g / cm ３ A cylindrical knitted fabric (hereinafter referred to as substrate A) was obtained. The polypropylene of the island component corresponds to the reinforcing material.

[0097] (Preparation of Substrate B) Polypropylene fibers having a single fineness of 16 dtex were spun under spinning conditions of a spinning speed of 1200 m / min, and 10 of these fibers were bundled to obtain a multifilament having a total fineness of 160 dtex. The obtained multifilament was knitted into a weight per unit area of ​​50 g / m using a cylindrical knitting machine in the same manner as in the preparation of Substrate 1. ２ The knitted fabric was made as follows.

[0098] Further, polysulfone pellets were dissolved in chloroform to prepare a 20% by mass polysulfone solution. The knitted fabric was immersed in the solution for 1 hour. After the impregnation, the substrate was pulled out and air-dried to obtain a polysulfone solution having a mass per unit area of ​​65 g / m. ２ A substrate B made of polypropylene coated with 23 wt % of polysulfone based on the total weight was obtained. The polypropylene fibers correspond to a reinforcing material.

[0099] (Preparation of Substrate C) 20 g of atactic polystyrene pellets were dissolved in 200 g of dichloromethane to prepare a polymer solution. 300 mL of ion-exchanged water was placed in a 500 mL separable flask, and 0.5 g of polyvinyl alcohol (PVA) was added. The PVA solution was prepared by stirring at 300 rpm using a three-one motor. Next, 4 mL of the polymer solution was filled into a 5 mL syringe, and the attached needle (inner diameter 0.23 mm) was poured into the PVA solution being stirred with a three-one motor. The polymer solution was discharged over 5 minutes and reprecipitated to obtain particles. The resulting particles were stirred for 1 hour. Only the resulting particles were sucked up with a pipette and collected in a centrifuge tube. The operation of discharging the polymer solution and collecting particles from the remaining PVA solution was repeated until the polymer solution was completely used. The particles collected in the centrifuge tube were vacuum-dried for 24 hours to obtain 17 g of substrate C composed of polystyrene particles. The particle diameter of the resulting particles was 35 μm. The substrate C does not include a reinforcing material.

[0100] (Preparation of Biological Component Treatment Material 1) 2.3 g of NMCA was added to a mixed solution of 31 g of nitrobenzene and 31 g of 98 wt% sulfuric acid, and the mixture was stirred at 10°C until the NMCA dissolved to prepare an NMCA solution. Next, 0.2 g of PFA was added to a mixed solution of 2.0 g of nitrobenzene and 2.0 g of 98 wt% sulfuric acid, and the mixture was stirred at 20°C until the PFA dissolved to prepare a PFA solution. 4.2 g of the PFA solution cooled to 5°C was mixed with 64.3 g of the NMCA solution, and the mixture was stirred for 5 minutes. 1 g of substrate A was then added to the resulting solution and impregnated for 2 hours. After the impregnation, substrate A was immersed in 200 mL of nitrobenzene at 0°C to stop the reaction, and the nitrobenzene adhering to substrate A was washed with methanol.

[0101] 0.24 g of tetraethylenepentamine (hereinafter referred to as TEPA) and 2.1 g of triethylamine were dissolved in 51 g of DMSO, and the substrate A, which had been washed with methanol, was added as it was and impregnated for 3 hours at 40° C. The substrate A was filtered on a glass filter and washed with 500 mL of DMSO.

[0102] To 47 g of DMSO that had been dehydrated and dried over activated molecular sieves 3A in advance, 0.075 g of parachlorophenyl isocyanate (hereinafter referred to as p-Cl) was added under a nitrogen atmosphere and heated to 30°C, and the washed substrate A was immersed in the entire amount for 1 hour. Substrate A was then filtered off on a glass filter to obtain a biological component processing material 1.

[0103] (Preparation of Biological Component Processing Material 2) Biological component processing material 2 was obtained in the same manner as in preparation of biological component processing material 1, except that 2.3 g of NMCA was changed to 1.725 g of NMCA and 0.575 g of NMA.

[0104] (Preparation of Biological Component Processing Material 3) Biological component processing material 3 was obtained using the same method as for preparing biological component processing material 1, except that 2.3 g of NMCA was replaced by 1.725 g of NMCA and 0.575 g of NMA, the amount of TEPA added was changed from 0.24 g to 0.48 g, and the amount of p-Cl added was changed from 0.075 g to 0.15 g.

[0105] (Preparation of Biological Component Processing Material 4) Biological component processing material 4 was obtained using the same method as for preparing biological component processing material 1, except that 2.3 g of NMCA was replaced by 1.725 g of NMCA and 0.575 g of NMA, the amount of TEPA added was changed from 0.24 g to 0.96 g, and the amount of p-Cl added was changed from 0.075 g to 0.3 g.

[0106] (Preparation of Biological Component Processing Material 5) Biological component processing material 5 was obtained in the same manner as in preparation of biological component processing material 1, except that 2.3 g of NMCA was changed to 1.15 g of NMCA and 1.15 g of NMA.

[0107] (Preparation of Biological Component Processing Material 6) Biological component processing material 6 was obtained using the same method as for preparing biological component processing material 1, except that 2.3 g of NMCA was replaced with 2.3 g of NMA. Note that, because NMCA, which is a partial structure of the first ligand, was not added to biological component processing material 6, the first ligand was not bound thereto.

[0108] (Preparation of Biological Component Processing Material 7) Biological component processing material 7 was obtained using the same method as for preparing biological component processing material 1, except that 2.3 g of NMCA was replaced with 0.575 g of NMCA and 1.725 g of NMA, 0.24 g of TEPA was replaced with 0.48 g of PEI, and the amount of p-Cl added was changed from 0.075 g to 0.15 g.

[0109] (Preparation of Biological Component Processing Material 8) Biological component processing material 8 was obtained in the same manner as in preparation of biological component processing material 1, except that 2.3 g of NMCA was changed to 2.3 g of NMCA and 0.46 g of NMBA.

[0110] (Preparation of Biological Component Processing Material 9) Biological component processing material 9 was obtained in the same manner as in preparation of biological component processing material 1, except that 2.3 g of NMCA was changed to 2.3 g of NMCA and 0.46 g of NMPA.

[0111] (Preparation of Biological Component Processing Material 10) A biological component processing material 10 was obtained using the same method as for preparing the biological component processing material 2, except that the substrate A was changed to the substrate B.

[0112] (Preparation of biocomponent processing material 11) Biocomponent processing material 11 was obtained in the same manner as biocomponent processing material 1, except that 0.24 g of TEPA was replaced with 0.48 g of PEI and the amount of p-Cl added was changed from 0.075 g to 0.15 g.

[0113] (Preparation of Biological Component Processing Material 12) A biological component processing material 12 was obtained in the same manner as in the preparation of the biological component processing material 1, except that 2.3 g of NMCA was replaced with 2.3 g of N-(hydroxymethyl)nicotinamide and 1.15 g of NMA.

[0114] (Preparation of Biological Component Processing Material 13) 1.7 g of NMCA was added to a mixed solution of 31 g of nitrobenzene and 31 g of 98 wt% sulfuric acid, and the mixture was stirred at 10°C until the NMCA dissolved, to obtain an NMCA solution. Next, 0.6 g of PFA was added to a mixed solution of 4.0 g of nitrobenzene and 4.0 g of 98 wt% sulfuric acid, and the mixture was stirred at 20°C until the PFA dissolved. 0.6 g of 2-hydroxyacetamide was then added and allowed to react to obtain a 2-hydroxy-N-(hydroxymethyl)acetamide / PFA mixed solution. 9.2 g of the 2-hydroxy-N-(hydroxymethyl)acetamide / PFA solution cooled to 5°C was mixed with 63.7 g of the NMCA solution, and the mixture was stirred for 5 minutes. 1 g of substrate A was then added to the resulting solution and impregnated for 2 hours. After the impregnation, substrate A was immersed in 200 mL of nitrobenzene at 0°C to stop the reaction, and the nitrobenzene adhering to substrate A was then washed with methanol.

[0115] 0.24 g of TEPA and 2.1 g of triethylamine were dissolved in 51 g of DMSO, and the substrate A, which had been washed with methanol, was added as it was and impregnated for 3 hours at 40° C. The substrate A was filtered on a glass filter and washed with 500 mL of DMSO.

[0116] 0.075 g of p-Cl was added to 47 g of DMSO that had been dehydrated and dried in advance using activated molecular sieves 3A under a nitrogen atmosphere, and the mixture was heated to 30°C, and the washed substrate A was immersed in the entire amount for 1 hour. Substrate A was then filtered off on a glass filter to obtain a biological component processing material 13.

[0117] (Preparation of Biological Component Processing Material 14) A biological component processing material 14 was obtained using the same method as for preparing the biological component processing material 13, except that 2-hydroxyacetamide was replaced with N-hydroxyoctanamide.

[0118] (Preparation of Biological Component Processing Material 15) A biological component processing material 15 was obtained using the same method as for preparing the biological component processing material 13, except that 2-hydroxyacetamide was replaced with 2-(4-butoxyphenyl)acetohydroxamic acid.

[0119] (Preparation of Biological Component Processing Material 16) Biological component processing material 16 was obtained in the same manner as in preparation of biological component processing material 1, except that substrate A was replaced with substrate C, and NMCA 2.3 g was replaced with NMCA 1.725 g and NMA 0.575 g.

[0120] <Evaluation Method> The evaluation method for the biological component treatment material is described below.

[0121] (Measurement of the weight proportion of water-insoluble material contained in the biocomponent treatment material) Approximately 5.0 g of the biocomponent treatment material was cut off and accurately weighed. The biocomponent treatment material and 100 mL of xylene were added to a 200 mL recovery flask and refluxed at 160°C for 24 hours to remove the polypropylene added as a reinforcing material. 2 L of xylene heated to 100°C was quickly added to the refluxed solution, and the insoluble components were directly filtered out using filter paper. The solution was washed with methanol and left to stand in a dryer at 80°C for 48 hours to obtain the water-insoluble material. The obtained water-insoluble material was dried, and the weight of the water-insoluble material was measured, and the weight proportion of the water-insoluble material was obtained using Equation 1. Weight proportion of water-insoluble material contained in the biocomponent treatment material = weight of water-insoluble material (g) / weight of biocomponent treatment material (g) ...Equation 1

[0122] (Measurement of Amino Group Content in the First Ligand of a Water-Insoluble Material Contained in a Biological Component Treatment Material) 1.0 g of the biological component treatment material and 50 mL of 6 M sodium hydroxide solution were added to a polypropylene container and stirred for 30 minutes. The biological component treatment material was then filtered using filter paper. The filtered biological component treatment material was then added to 50 mL of ion-exchanged water, stirred for 30 minutes, and filtered using filter paper. This process of adding the biological component treatment material to ion-exchanged water and filtering was repeated until the pH of the ion-exchanged water containing the biological component treatment material reached 7, yielding a desalted biological component treatment material. The desalted biological component treatment material was then allowed to stand at 80°C under normal pressure for 48 hours, after which 1.0 g of the biological component treatment material and 30 mL of 0.1 M hydrochloric acid were added to a polypropylene container and stirred for 10 minutes. After stirring, 5 mL of the solution was removed and transferred to another polypropylene container. Next, 0.1 mL of 0.1 M sodium hydroxide solution was added dropwise to the resulting solution. After the addition, the solution is stirred for 10 seconds using a vortex mixer, and the pH of the solution is measured. The stirring and pH measurement after the addition are repeated 50 times in the same manner. The amount of sodium hydroxide solution added when the pH of the solution exceeds 8.5 is taken as the titration amount. The value obtained using the titration amount and the following formula 2 is rounded to one decimal place to calculate the amino group content in the first ligand per 1 g of dry weight of the water-insoluble material. Amino group content in the first ligand per 1 g of water-insoluble material (mmol / g) = [{amount of 0.1 M hydrochloric acid added (30 mL) / amount of hydrochloric acid removed (5 mL)} × {amount of neutralization solution (5 mL) - titration amount (mL)} × concentration of sodium hydroxide solution (0.1 M)] / {dry weight of added biological component processing material (1 g) × weight proportion of water-insoluble material contained in biological component processing material} ...Formula 2

[0123] (Measurement of the content of the second ligand of the water-insoluble material contained in the biological component treatment material) The content of the second ligand of the water-insoluble material contained in the biological component treatment material is measured by hydrolyzing the biological component treatment material and quantifying the amount of eluate derived from the eluted second ligand. Details are described below.

[0124] When the biocomponent processing material is in the form of a knitted fabric (when the biocomponent processing material is made using base material A or B), the biocomponent processing material should be 2 cm thick per piece. ２Six pieces of the same size are cut out, and if the biocomponent processing material is in particulate form (in the case of a biocomponent processing material prepared using substrate C), 1 mL of the biocomponent processing material is drawn up with a pipette. The collected biocomponent processing material is dried and its dry weight is measured. The biocomponent processing material after its dry weight measurement is then placed in a pressure-resistant glass bottle and heated at 110°C for 20 hours. After 20 hours of heating, 0.99 mL of the solution in the pressure-resistant glass bottle is collected and 0.01 mL of phenol is added as an internal standard to prepare a measurement solution. 6 M aqueous sodium hydroxide solution is added in 0.05 mL increments until the pH exceeds 7.0. 0.1 mL of the resulting solution is removed and mixed with 10 mg of sodium trimethylsilylpropanesulfonate (hereinafter referred to as DSS) and 0.6 mL of heavy water. １ H-NMR is measured. The peak derived from DSS is set to 0 ppm, and the integral value of the phenol peak (δ = 7.2-7.3 ppm) is normalized to 2 to calculate the integral value of the peak derived from the second ligand. The value obtained using Equation 3 is rounded to one decimal place to calculate the content of the second ligand per gram of dry weight of the water-insoluble material. Content of second ligand per gram of dry weight of water-insoluble material (mmol / g) = [{integral value of the peak derived from the second ligand / number of protons of the peak derived from the second ligand} × phenol concentration in the measurement solution (0.112 mol / L) × volume of degradation solution (3 mL)] / {weight (g) of added biological component processing material × weight proportion of water-insoluble material contained in the biological component processing material} ...Equation 3

[0125] (Measurement of IL-6 adsorption rate of biological component processing material) To confirm the IL-6 adsorption capacity of the biological component processing material, the biological component processing material was immersed in a liquid containing IL-6 for a predetermined period of time, then removed, and the IL-6 adsorption rate was measured from the difference in the amount of IL-6 in the liquid before and after impregnation. The measurement method is shown below.

[0126] When the biocomponent processing material is in knitted form, four disks with a diameter of 6 mm are cut out from the biocomponent processing material, the thickness is measured with a micrometer, the total volume of the four disks is calculated, and the entire amount is placed in a polypropylene container. When the biocomponent processing material is in particulate form, 0.05 mL of the biocomponent processing material is sucked up with a pipette and the entire amount is placed in a polypropylene container. FBS adjusted to an IL-6 concentration of 2000 pg / mL is added to this container, and the total volume of the biocomponent processing material is 1 cm. ３ The solution is added to the above container at a solid-liquid ratio of 30 mL to 1 mL. After mixing by inversion for 2 hours in a 37°C incubator, the IL-6 concentration in the FBS is measured by ELISA. The IL-6 adsorption rate is calculated by rounding off the value obtained from the IL-6 concentration before and after mixing by inversion using the following formula 4. A microplate reader (Spectra Max M5, manufactured by Molecular Devices) is used to measure absorbance, with a measurement wavelength of 450 nm and a reference wavelength of 595 nm, after a blank measurement is performed in advance. IL-6 adsorption rate (%) = 100 × {IL-6 concentration before mixing by inversion (pg / mL) - IL-6 concentration after mixing by inversion (pg / mL)} / IL-6 concentration before mixing by inversion (pg / mL) ... formula 4

[0127] (Measurement of the rate of decrease in IL-6 adsorption capacity of a biocomponent processing material before and after autoclaving) 10 mL of physiological saline solution was added to a 15 mL glass vial per 1 g of dry weight of the biocomponent processing material, and the biocomponent processing material was autoclaved using an autoclave (manufactured by Tony Seiko Co., Ltd.) at a sterilization temperature of 117°C for a sterilization time of 105 minutes. The IL-6 adsorption capacity of the biocomponent processing material after autoclaving was determined in the same manner as in the above-mentioned "Measurement of the IL-6 adsorption capacity of a biocomponent processing material," except that the obtained autoclaved biocomponent processing material was used. The rate of decrease in IL-6 adsorption capacity of the biocomponent processing material after autoclaving was calculated by rounding the value obtained using Equation 5 to one decimal place. Rate of decrease in IL-6 adsorption capacity (%) = 100 × {IL-6 adsorption capacity (%) - IL-6 adsorption capacity after autoclaving (%)} / IL-6 adsorption capacity (%) Equation 5

[0128] (Measurement of IL-8 adsorption rate for biocomponent processing material) The IL-8 adsorption rate was determined by rounding off the value obtained using Equation 6 to one decimal place in the same manner as in the above-mentioned "Measurement of IL-6 adsorption rate for biocomponent processing material" except for changing IL-6 to IL-8. IL-8 adsorption rate (%) = 100 × {IL-8 concentration before mixing by inversion (pg / mL) - IL-8 concentration after mixing by inversion (pg / mL)} / IL-8 concentration before mixing by inversion (pg / mL) ...Equation 6

[0129] (Measurement of the rate of decrease in IL-8 adsorption performance of a biological component processing material before and after autoclave sterilization) The IL-8 adsorption performance of a biological component processing material after autoclave sterilization is determined using the same method as for measuring the rate of decrease in IL-6 adsorption performance of a biological component processing material before and after autoclave sterilization, except that IL-6 is replaced with IL-8. The value obtained using Equation 7 is then rounded to the nearest whole number to obtain the rate of decrease in IL-8 adsorption performance. IL-8 adsorption performance decrease rate (%) = 100 × {IL-8 adsorption rate (%) - IL-8 adsorption rate after autoclave sterilization (%)} / IL-8 adsorption rate (%) ... Equation 7

[0130] (Measurement of pH of filling solution after autoclaving of biocomponent processing material) 10 mL of physiological saline solution was added to a 15 mL glass vial for every 1 g of dry weight of biocomponent processing material, and the biocomponent processing material was autoclaved using an autoclave (manufactured by Tony Seiko Co., Ltd.) at a sterilization temperature of 117°C for a sterilization time of 105 minutes. The biocomponent processing material was removed from the glass vial, and the pH of the remaining solution was measured. The value obtained was rounded to one decimal place and used as the pH of the filling solution after autoclaving.

[0131] <Evaluation of biological component processing materials>

[0132] (Measurement of weight proportion of water-insoluble material contained in biological component treatment materials 1 to 16) The weight proportion of water-insoluble material contained in each of biological component treatment materials 1 to 16 was obtained according to the procedure of "Measurement of weight proportion of water-insoluble material contained in biological component treatment material" described above.

[0133] (Measurement of Amino Group Content in First Ligand of Water-Insoluble Material Contained in Biological Component Treatment Materials 1 to 16) The amino group content in the first ligand of the water-insoluble material contained in each of Biological Component Treatment Materials 1 to 16 was determined according to the procedure described above in "Measurement of Amino Group Content in First Ligand of Water-Insoluble Material Contained in Biological Component Treatment Materials." The measurement results for Biological Component Treatment Materials 1 to 10 are shown in Table 1, and the measurement results for Biological Component Treatment Materials 11 to 16 are shown in Table 2.

[0134] (Measurement of the content of the second ligand of the water-insoluble material contained in biological component treatment materials 1 to 16) For biological component treatment materials 1 and 11, the compound from which the second ligand is derived was not added, and therefore the content of the second ligand was set to 0 mmol / g. For biological component treatment materials 2 to 7, 9, 10, 12, and 16, the content of the second ligand of the water-insoluble material contained in each of biological component treatment materials 2 to 7, 9, 10, 12, and 16 was obtained according to the procedure described above in "Measurement of the content of the second ligand of the water-insoluble material contained in biological component treatment material," with the peak derived from the second ligand being 1.9-2.1 ppm and the number of protons of the peak derived from the second ligand being 3. For biological component treatment material 8, the content of the second ligand of the water-insoluble material contained in biological component treatment material 8 was obtained according to the procedure described above in "Measurement of the content of the second ligand of the water-insoluble material contained in biological component treatment material," with the peak derived from the second ligand being 8.0-8.2 ppm and the number of protons of the peak derived from the second ligand being 2. For biological component processing material 13, the content of the second ligand of the water-insoluble material contained in biological component processing material 13 was obtained according to the procedure of "Measuring the content of the second ligand of the water-insoluble material contained in the biological component processing material" described above, with the peak derived from the second ligand being at 4.2-4.3 ppm and the number of protons of the peak derived from the second ligand being 2. For biological component processing material 14, the content of the second ligand of the water-insoluble material contained in biological component processing material 14 was obtained according to the procedure of "Measuring the content of the second ligand of the water-insoluble material contained in the biological component processing material" described above, with the peak derived from the second ligand being at 0.7-0.9 ppm and the number of protons of the peak derived from the second ligand being 3. For biological component processing material 15, the content of the second ligand of the water-insoluble material contained in biological component processing material 15 was obtained according to the procedure of "Measuring the content of the second ligand of the water-insoluble material contained in the biological component processing material" described above, with the peak derived from the second ligand being at 7.2-8.1 ppm and the number of protons of the peak derived from the second ligand being 4. The results for biological component processing materials 1 to 10 are shown in Table 1, and the results for biological component processing materials 11 to 16 are shown in Table 2.

[0135] Example 1 For biological component processing material 2, the IL-6 adsorption rate, IL-6 adsorption rate reduction rate, IL-8 adsorption rate, IL-8 adsorption rate, IL-8 adsorption rate, and IL-8 adsorption rate reduction rate, as well as the pH of the filling solution after autoclave sterilization, were measured according to the procedures described above in "Measurement of IL-6 adsorption rate for biological component processing material," "Measurement of the rate of decrease in IL-6 adsorption performance for biological component processing material before and after autoclave sterilization," "Measurement of the rate of decrease in IL-8 adsorption performance for biological component processing material before and after autoclave sterilization," and "Measurement of the pH of the filling solution after autoclave sterilization of biological component processing material." The results are shown in Table 3.

[0136] For biological component processing material 3, the IL-6 adsorption rate, the IL-6 adsorption performance reduction rate, the IL-8 adsorption rate, the IL-8 adsorption performance reduction rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0137] For the biological component processing material 4, the IL-6 adsorption rate, the IL-6 adsorption performance reduction rate, the IL-8 adsorption rate, the IL-8 adsorption performance reduction rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0138] For biological component processing material 5, the IL-6 adsorption rate, the IL-6 adsorption performance reduction rate, the IL-8 adsorption rate, the IL-8 adsorption performance reduction rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0139] For the biological component processing material 7, the IL-6 adsorption rate, the IL-6 adsorption performance reduction rate, the IL-8 adsorption rate, the IL-8 adsorption performance reduction rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0140] For biological component processing material 8, the IL-6 adsorption rate, the IL-6 adsorption performance decline rate, the IL-8 adsorption rate, the IL-8 adsorption performance decline rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0141] For the biological component processing material 9, the IL-6 adsorption rate, the IL-6 adsorption performance reduction rate, the IL-8 adsorption rate, the IL-8 adsorption performance reduction rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0142] For the biological component processing material 10, the IL-6 adsorption rate, the IL-6 adsorption performance decline rate, the IL-8 adsorption rate, the IL-8 adsorption performance decline rate, and the pH of the filling solution after autoclave sterilization were measured in the same manner as in Example 1. The results are shown in Table 3.

[0143] For the biological component processing material 13, the IL-6 adsorption rate, the IL-6 adsorption performance decline rate, the IL-8 adsorption rate, the IL-8 adsorption performance decline rate, and the pH of the filling solution after autoclave sterilization were measured in the same manner as in Example 1. The results are shown in Table 3.

[0144] For the biological component processing material 16, the IL-6 adsorption rate, the IL-6 adsorption performance decline rate, the IL-8 adsorption rate, the IL-8 adsorption performance decline rate, and the pH of the filling solution after autoclave sterilization were measured in the same manner as in Example 1. The results are shown in Table 3.

[0145] Comparative Example 1 For biological component processing material 1, the IL-6 adsorption rate, IL-6 adsorption performance decline rate, IL-8 adsorption rate, IL-8 adsorption performance decline rate, and pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0146] For biological component processing material 6, the IL-6 adsorption rate, the IL-6 adsorption performance decline rate, the IL-8 adsorption rate, the IL-8 adsorption performance decline rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0147] For the biological component processing material 11, the IL-6 adsorption rate, the IL-6 adsorption performance reduction rate, the IL-8 adsorption rate, the IL-8 adsorption performance reduction rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0148] Comparative Example 4 For the biological component processing material 12, the IL-6 adsorption rate, the IL-6 adsorption performance reduction rate, the IL-8 adsorption rate, the IL-8 adsorption performance reduction rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0149] Comparative Example 5 For the biological component processing material 14, the IL-6 adsorption rate, the IL-6 adsorption performance decline rate, the IL-8 adsorption rate, the IL-8 adsorption performance decline rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0150] Comparative Example 6 For the biological component processing material 15, the IL-6 adsorption rate, the IL-6 adsorption performance reduction rate, the IL-8 adsorption rate, the IL-8 adsorption performance reduction rate, and the pH of the filling solution after autoclaving were measured in the same manner as in Example 1. The results are shown in Table 3.

[0151]

[0152]

[0153] In Tables 1 and 2, "content of amino groups in first ligand" means the content of amino groups in the first ligand per gram of dry weight of the water-insoluble material, and "content of second ligand" means the content of second ligand per gram of dry weight of the water-insoluble material.

[0154]

[0155] The results of Comparative Examples 1 and 3 in Table 3 indicate that conventional biological component processing materials exhibit high rates of decline in IL-6 and IL-8 adsorption performance due to autoclaving, making them unsuitable for autoclaving. In contrast, the results of Examples 1 to 5 indicate that the inclusion of a second ligand in addition to a first ligand can suppress the decline in IL-6 and IL-8 adsorption performance due to autoclaving, and furthermore, can suppress the acidification of the pH of the filling solution after autoclaving. Furthermore, the results of Example 5 and Comparative Example 4 indicate that these effects are exhibited within the range of the chemical structure represented by general formula (I), regardless of the chemical structure of the first ligand. Furthermore, the results of Examples 6 and 7 and Comparative Examples 5 and 6 indicate that these effects are exhibited within the range of the chemical structure represented by general formula (IIa) or (IIb), regardless of the chemical structure of the second ligand. Furthermore, the results of Example 8 indicate that these effects are exhibited regardless of the substrate material.

[0156] The biological component processing material of the present invention can be suitably used as an adsorption material in biological component processing columns intended to treat unwanted biological components, particularly in blood purification columns for treating inflammatory diseases.

Claims

1. A material for treating a biological component, comprising a water-insoluble material including a first ligand having an amino group and an amide group, a second ligand having a hydrocarbon group and an amide group which may be substituted with a hydroxy group, and a substrate, wherein the first ligand is bound to the substrate by a structure represented by the following general formula (I), and the second ligand is bound to the substrate by a structure represented by the following general formula (IIa) or (IIb). [In the formula, the wavy line represents the bonding position to the substrate, and X represents a chemical structure that contains an amino group as a partial structure and may also contain a phenyl group.] [In the formula, the wavy line represents the bonding position to the substrate, A represents an alkyl group having 1 to 6 carbon atoms or a phenyl group which may be substituted with a hydroxy group, and n represents an integer of 1 to 6.] 2. The material for treating a biological component according to claim 1, wherein the content of said second ligand is 0.3 to 5.0 mmol per 1 g of the dry weight of said water-insoluble material.

3. The material for treating a biological component according to claim 1 or 2, wherein the content of amino groups in said first ligand is 1.0 to 4.0 mmol per gram of dry weight of said water-insoluble material.

4. A column for treating a biological component comprising the biological component treating material according to any one of claims 1 to 3.

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