Inflammatory mediator absorbent

Abstract

To provide an inflammatory mediator absorbent capable of efficiently absorbing an inflammatory mediator, a manufacturing method of the inflammatory mediator absorbent, and a use of the inflammatory mediator absorbent.SOLUTION: An inflammatory mediator absorbent according to the present invention includes a water-insoluble carrier, and a ligand with respect to an inflammatory mediator. The ligand has an aromatic hydrocarbon group having two or more phenolic hydroxyl groups and the ligand is bonded to the water-insoluble carrier.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an inflammatory mediator adsorbent capable of efficiently adsorbing inflammatory mediators, a method for producing the same, and uses thereof. [Background technology]

[0002] Inflammatory mediators are a general term for chemicals involved in inflammatory responses and are involved in the body's defense responses, but they can also cause excessive tissue damage and excessive immunosuppression. Furthermore, inflammatory mediators are primarily intended to act in local immune responses; however, if their blood levels rise, they can actually act as endotoxin-inducing factors, leading to conditions such as sepsis. For this reason, various antibody drugs against inflammatory mediators have been developed, but systems have also been developed to rapidly reduce blood levels of inflammatory mediators by drawing blood, selectively adsorbing and removing inflammatory mediators, and then returning the purified blood.

[0003] For example, Patent Documents 1 and 2 disclose adsorbents for TNFα and interleukin, respectively, in which a hydrophobic compound with a log P value of 2.50 or more, such as cetylamine, is immobilized on a water-insoluble porous carrier.

[0004] Furthermore, Patent Document 3 discloses an adsorption carrier for hepatitis treatment, in which a functional group having an amino group is introduced onto the surface of a water-insoluble carrier, and which adsorbs interleukins and the like. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Publication No. 8-257115 [Patent Document 2] Japanese Patent Application Publication No. 8-257398 [Patent Document 3] Japanese Patent Application Laid-Open No. 2012-5827 Summary of the Invention [Problem to be solved by the invention]

[0006] As mentioned above, various adsorbents have been developed to adsorb and remove inflammatory mediators from blood, and these mainly utilize hydrophobic interactions or electrostatic bonds such as amino groups. However, in recent years, infectious diseases have become a problem, and the risk of systemic inflammatory response syndrome has increased, so there is a demand for adsorbents that can adsorb and remove inflammatory mediators more efficiently. Therefore, an object of the present invention is to provide an inflammatory mediator adsorbent capable of efficiently adsorbing inflammatory mediators, a method for producing the same, and uses thereof. [Means for solving the problem]

[0007] The present inventors have conducted extensive research to solve the above problems, and as a result have found that a ligand having an aromatic hydrocarbon group with two or more phenolic hydroxyl groups can efficiently adsorb inflammatory mediators, thereby completing the present invention. The present invention will now be described.

[0008] [1] A pharmaceutical composition comprising a water-insoluble carrier and a ligand for an inflammatory mediator, the ligand has an aromatic hydrocarbon group having two or more phenolic hydroxyl groups, An inflammatory mediator adsorbent, characterized in that the ligand is bound to the water-insoluble carrier. [2] The inflammatory mediator adsorbent according to [1], wherein at least two of the phenolic hydroxyl groups are adjacent to each other on the aromatic hydrocarbon group. [3] The inflammatory mediator adsorbent according to [1] or [2] above, wherein the aromatic hydrocarbon group is a catechol group. [4] The ligand is attached to the water-insoluble carrier by -N(R 1 )-group (wherein, R 1 is H or C 1-6The inflammatory mediator adsorbent according to any one of the above [1] to [3], wherein the inflammatory mediator is covalently bonded via a group (representing an alkyl group). [5] The inflammatory mediator adsorbent according to any one of [1] to [4], wherein the inflammatory mediator is one or more inflammatory mediators selected from IL-6, TNFα, IL-10, IL-8, calprotectin, and HMGB1. [6] The adsorbent for inflammatory mediators according to any one of [1] to [5] above, wherein the water-insoluble carrier comprises a cellulose derivative.

[0009] [7] A housing comprising: the housing having a liquid inlet and an outlet; An inflammatory mediator adsorber, characterized in that the housing is filled with the inflammatory mediator adsorbent according to any one of [1] to [6] above.

[0010] [8] A method for producing an inflammatory mediator adsorbent, comprising: A method characterized by comprising a step of covalently bonding a ligand to a water-insoluble support by reacting a ligand precursor compound having an aromatic hydrocarbon group having two or more phenolic hydroxyl groups with the support. [9] The method according to [8] above, wherein the ligand precursor compound is L-3,4-dihydroxyphenylalanine or dopamine. [Effects of the Invention]

[0011] The adsorbent of the present invention has a ligand having an aromatic hydrocarbon group with two or more phenolic hydroxyl groups. It is believed that such a ligand exhibits high affinity for inflammatory mediators, likely due to both hydrophobic interactions derived from the aromatic hydrocarbon group and hydrogen bonding through two or more phenolic hydroxyl groups per aromatic hydrocarbon group. The adsorbent of the present invention can efficiently adsorb and remove IL-6 and TNFα in particular. IL-6 and TNFα are particularly problematic among inflammatory mediators, and their overexpression is particularly problematic, as they promote sepsis and also promote the differentiation and activation of osteoclast-like cells, which are the cause of rheumatoid arthritis. Therefore, the present invention is industrially highly advantageous as a technology for removing inflammatory mediators from biological fluids such as blood. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 is a schematic diagram showing an example of an inflammatory mediator adsorber according to the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0013] The present invention will be described in more detail below with reference to specific examples, but the present invention is not limited to the following specific examples. The inflammatory mediator adsorbent according to the present invention comprises a water-insoluble carrier and a ligand for an inflammatory mediator.

[0014] The adsorbent according to the present invention adsorbs inflammatory mediators. Inflammatory mediators are physiologically active substances released from leukocytes, mast cells, macrophages, and the like that infiltrate damaged tissues or inflammatory sites, and are a general term for endogenous substances that initiate and maintain inflammatory responses in the body. Examples of inflammatory mediators include inflammatory cytokines such as IL-1β, IL-6, IL-8, IL-10, IL-18, TNFα, and HMGB1; pain-inducing substances such as bradykinin, serotonin, and histamine; prostanoids such as prostaglandins; complement components such as C3a and C5a; leukotrienes, platelet-activating factor (PAF), elastase, collagenase, and calprotectin.

[0015] In particular, the adsorption targets of the present invention include inflammatory cytokines that promote inflammatory responses, particularly IL-6 and TNFα, which are involved in excessive inflammatory responses and immune system dysregulation, leading to conditions such as rheumatoid arthritis. IL-8 secretion is increased by oxidative stress, leading to local inflammation by recruiting inflammatory cells and further promoting inflammation. IL-8 has also been shown to be associated with angiogenesis, obesity, promotion of cancer cell division and metastasis, and the risk of schizophrenia in pregnant women. Furthermore, alarmins such as HMGB1 and calprotectin, which are released from cells as a result of the invasion and destruction of self-tissues caused by released cytokines, have been shown to contribute to the exacerbation of inflammatory conditions such as septic shock and SIRS. Although IL-10 is an anti-inflammatory cytokine, its overexpression can excessively suppress the immune system and lead to secondary bacterial and fungal infections. Therefore, in pathological conditions in which excessive cytokines are produced in the body, such as sepsis, it is necessary to reduce not only inflammatory cytokines and alarmins but also anti-inflammatory cytokines in a balanced manner. Therefore, for the sake of convenience, in the present disclosure, inflammatory mediators include not only inflammatory cytokines and alarmins but also anti-inflammatory cytokines.

[0016] The material of the water-insoluble carrier is not particularly limited as long as it is insoluble in water, but examples include polysaccharides, polystyrene, styrene-divinylbenzene copolymers, polyacrylamide, poly(meth)acrylic acid, poly(meth)acrylic acid esters, and derivatives thereof. These may have a coating layer of a polymer material having a hydroxyl group, such as polyhydroxyethyl methacrylate, or a graft copolymer such as a copolymer of a monomer having a polyethylene oxide chain and another polymerizable monomer. Among these, polysaccharides are preferred because they have hydroxyl groups on the carrier surface and active groups can be easily introduced via the hydroxyl groups.

[0017] Polysaccharides are preferred because they are easily available industrially and are highly safe for living organisms. There are no particular limitations on the polysaccharides that can be used in the water-insoluble carrier of the present invention, but examples include agarose, cellulose, dextrin, chitosan, chitin, and derivatives thereof.

[0018] Furthermore, the material of the water-insoluble carrier of the present invention preferably contains cellulose and / or a cellulose derivative. Porous carriers containing cellulose or cellulose derivatives have relatively high mechanical strength and are tough, so they are less likely to break or produce fine particles, and when packed in a column, they are relatively resistant to compaction even when liquid is passed through them at high linear velocity. Furthermore, from the standpoints of strength and cost, cellulose is the most preferred material for the water-insoluble carrier of the present invention.

[0019] The shape of the water-insoluble carrier is not particularly limited, but may be, for example, spherical, bead-like, monolithic, fibrous, or membrane-like (including hollow fibers). The molecular weight of the material of the water-insoluble carrier is also not particularly limited, but preferably has an exclusion limit molecular weight of 1,000 or more and 1,000,000 or less. The exclusion limit molecular weight is more preferably 10,000 or more, even more preferably 20,000 or more, and more preferably 500,000 or less, even more preferably 100,000 or less.

[0020] The size of the water-insoluble carrier may be adjusted as appropriate, but for example, an average particle size of 20 μm or more and 1000 μm or less is preferred. If the average particle size of the water-insoluble carrier is 20 μm or more, compaction is unlikely to occur, and if it is 1000 μm or less, the surface area is sufficiently large, resulting in a sufficiently large amount of adsorption of inflammatory mediators. The average particle size of the water-insoluble carrier is preferably 50 μm or more, more preferably 100 μm or more, even more preferably 200 μm or more, and preferably 800 μm or less, more preferably 600 μm or less. The average particle size of the water-insoluble carrier can be determined by measuring the diameter of each particle individually using image analysis software from a magnified photograph of the carrier taken with a stereoscopic microscope or the like, and then determining the particle size on a number basis.

[0021] The ligands of the present invention have an aromatic hydrocarbon group with two or more phenolic hydroxyl groups, and exhibit strong affinity for inflammatory mediators, probably due to a combination of hydrophobic interactions by the aromatic hydrocarbon group and hydrogen bonds by the phenolic hydroxyl groups.

[0022] Examples of the aromatic hydrocarbon group possessed by the ligand include C groups such as phenyl, naphthyl, indenyl, and biphenyl groups. 6-12 Examples include aromatic hydrocarbon groups, preferably a phenyl group or a naphthyl group, and more preferably a phenyl group.

[0023] Two or more phenolic hydroxyl groups on one aromatic hydrocarbon group are preferably located at the p- or o-position relative to each other on the phenyl group. Two phenolic hydroxyl groups located at the p- or o-position form a quinone structure. Such a quinone structure may improve affinity for inflammatory mediators. Furthermore, two phenolic hydroxyl groups located at the o-position relative to each other may improve affinity for inflammatory mediators. For the same reason, it is also preferable that at least two phenolic hydroxyl groups are adjacent on an aromatic hydrocarbon group other than a phenyl group. Here, "phenolic hydroxyl groups are adjacent" means that the carbon atoms substituted with phenolic hydroxyl groups are covalently bonded to each other.

[0024] The number of phenolic hydroxyl groups per aromatic hydrocarbon group is not particularly limited as long as it is substitutable, but can be, for example, 2 or more and 10 or less. The number is preferably 7 or less or 5 or less, more preferably 4 or less, and even more preferably 2 or 3.

[0025] Examples of aromatic hydrocarbon groups having two or more phenolic hydroxyl groups include catechol groups such as a 3,4-hydroxyphenyl group, a 3,4,6-trihydroxyphenyl group, a 1,8-dihydroxynaphthyl group, a 5,7-dihydroxynaphthyl group, a 5,8-dihydroxynaphthyl group, and a 6,7-dihydroxynaphthyl group.

[0026] In the inflammatory mediator adsorbent of the present invention, a ligand is bound to a water-insoluble carrier. The type of binding is not particularly limited, as long as it prevents the ligand from falling off the water-insoluble carrier to an extent that does not impair the effect, for example, when treating a liquid containing the inflammatory mediator to reduce the inflammatory mediator. Examples of the binding include covalent bonding, electrostatic bonding, and hydrogen bonding. Furthermore, at least a portion of the ligand may be polymerized with another ligand to coat at least a portion of the water-insoluble carrier.

[0027] The ligand is preferably covalently bonded to the water-insoluble carrier directly or indirectly via a linker group, and more preferably indirectly via a linker group. For example, an aromatic hydrocarbon group having a phenolic hydroxyl group is preferably covalently bonded to the water-insoluble carrier via a linker group. The linker group has the role of increasing the positional freedom of the ligand and facilitating the introduction of the aromatic hydrocarbon group into the water-insoluble carrier. The linker group is not particularly limited as long as it has such a function, but examples thereof include C 1-6 Alkylene group, divalent amino group (-N(R 1 )-, where R 1 is H (hydrogen atom) or C 1-6 Examples of the linked group include a C alkyl group (-O-), an ether group (-O-), a thioether group (-S-), a carbonyl group (-C(=O)-), a thionyl group (-C(=S)-), an ester group (-O-C(=O)- or -C(=O)-O-), an amide group (-NH-C(=O)- or -C(=O)-NH-), a urea group (-NH-C(=O)-NH-), a thiourea group (-NH-C(=S)-NH-), and a group in which two or more and five or less of these groups are linked together. Examples of the linked group include a C alkyl group having at one or both ends a group selected from the group consisting of a divalent amino group, an ether group, a thioether group, a carbonyl group, a thionyl group, an ester group, an amide group, a urea group, a thiourea group, a polyalkylene glycol group, and a polyvinyl alcohol group. 1-6Examples of the linker group include an alkylene group. The linker group may also have a substituent such as a carboxy group. Electrostatic bonding via a divalent amino group or carboxy group may further improve affinity for inflammatory mediators.

[0028] In this disclosure, "C 1-6 The term "alkyl group" refers to a linear or branched monovalent saturated aliphatic hydrocarbon group having 1 to 6 carbon atoms. Examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, t-butyl, n-pentyl, and n-hexyl. Preferably, C 1-4 alkyl group, more preferably C 1-2 It is preferably an alkyl group, and more preferably methyl. 1-6 The term "alkylene group" refers to a linear or branched divalent saturated aliphatic hydrocarbon group having 1 to 6 carbon atoms. Examples include methylene, ethylene, methylmethylene, n-propylene, methylethylene, n-butylene, methylpropylene, dimethylethylene, n-pentylene, and n-hexylene. Preferably, C 1-4 alkylene, more preferably C 3-4 It is alkylene.

[0029] The amount of ligand in the adsorbent of the present invention is not particularly limited as long as inflammatory mediators are adsorbed satisfactorily, but can be, for example, 10 μmol or more and 200 μmol or less per mL of sedimentation volume of the adsorbent. The ratio is preferably 15 μmol / mL or more or 20 μmol / mL or more, more preferably 30 μmol / mL or more or 40 μmol / mL or more, even more preferably 50 μmol / mL or more, and preferably 150 μmol / mL or less, more preferably 100 μmol / mL or less. The sedimentation volume of the adsorbent refers to, for example, the volume measured when approximately 5 mL of the adsorbent is placed in a 15 mL centrifuge tube and shaken until the adsorbent volume no longer decreases.

[0030] The method for producing the inflammatory mediator adsorbent according to the present invention will be described below.

[0031] 1. Activation process of water-insoluble carrier In this step, the surface of the raw material water-insoluble support is activated. This step is optional, and for example, when using a water-insoluble support on the surface of which active groups have been introduced, this step does not need to be performed.

[0032] Since polysaccharides have hydroxyl groups and poly(meth)acrylic acid has carboxyl groups, these functional groups can be activated or active groups can be introduced via these functional groups. Since polystyrene and the like do not have functional groups, functional groups can be introduced onto the surface by coating the raw water-insoluble support with a polymer material having hydroxyl groups, such as polyhydroxyethyl methacrylate, or a graft copolymer, such as a copolymer of a monomer having a polyethylene oxide chain and another polymerizable monomer.

[0033] For example, in the case of a water-insoluble support having hydroxyl groups on its surface, highly reactive epoxy groups may be introduced by reacting epihalohydrin such as epichlorohydrin with the hydroxyl groups. Furthermore, the introduced epoxy groups may be oxidized to formyl groups. Carboxy groups may be converted to acid halide groups using thionyl chloride or the like, or may be activated with a base, water-soluble carbodiimide, or the like.

[0034] 2. Ligand binding process In this step, a ligand precursor compound is reacted with the activated water-insoluble support to bind the ligand.

[0035] The ligand precursor compound has an aromatic hydrocarbon group with two or more phenolic hydroxyl groups, and can be reacted with the active groups of the activated water-insoluble support to covalently bond the desired ligand to the water-insoluble support. Examples of the ligand precursor compound include 3,4-dihydroxybenzylamine, dopamine, 4-(3-aminopropyl)-1,2-benzenediol, 3,4,6-trihydroxybenzylamine, 6-hydroxydopamine, 4-(3-aminopropyl)-1,2,6-benzenetriol, L-dopa (L-3,4-dihydroxyphenylalanine), and 6-hydroxy-L-dopa.

[0036] For example, in the case of a ligand precursor compound having an amino group (-NH), since the amino group has strong nucleophilicity, it reacts with an epoxy group and is supported on a water-insoluble support via the -NH- group. In addition, when the water-insoluble support has a formyl group on the surface, the ligand precursor compound having an amino group can be supported on a water-insoluble support via a reductive amination reaction to form a -N(R 1 A ligand precursor compound having an amino group can be covalently bonded to a water-insoluble support via an amide group.

[0037] The ligand is -N(R 1 Preferably, the compound is covalently bonded to the water-insoluble carrier via an —N(R )- group. 1 The -N(R)- group may further enhance the adsorption of inflammatory cytokines. 1 The phrase "covalently bonded to the water-insoluble carrier via an -N(R)- group" means that the ligand and the water-insoluble carrier are bonded to each other via an -N(R)- group. 1 The ligand and the water-insoluble carrier may be directly linked by an -N(R 1 This means that they may be attached by a linker group containing a - group.

[0038] After the ligand is introduced into the water-insoluble carrier, a conventional post-treatment may be carried out, for example, by washing with water, an acidic solution, a buffer solution, or the like.

[0039] The adsorbent according to the present invention has excellent adsorption capacity for inflammatory mediators, and can therefore be used as an adsorbent for adsorbing inflammatory mediators by filling a housing having an inlet and outlet for liquid with the adsorbent according to the present invention.

[0040] When at least a portion of the water-insoluble support is to be coated with a polymer of the ligand, the ligand may be polymerized on the raw water-insoluble support whose surface has been activated or on the raw water-insoluble support whose surface has not been activated.

[0041] An example of an adsorber is shown schematically in Figure 1. The adsorber in Figure 1 has a housing 6 filled with an adsorbent 3 according to the present invention. Filters 4 and 5 are attached to both ends of the housing 6, preventing the adsorbent 3 from leaking while allowing liquid to flow through the adsorbent 3. Covers 1b and 2b, which can be fixed to the housing 6 in a liquid-tight manner with screws or the like, are attached to the surfaces of the housing 6 on which the filters 4 and 5 are attached. Liquid can be supplied to the adsorbent 3 in the housing 6 through a liquid inlet 1a formed in these covers 1b and 2b, and liquid that has come into contact with the adsorbent 3 in the housing 6 can be discharged through an outlet 2a.

[0042] By using an adsorbent filled with the inflammatory mediator adsorbent according to the present invention, it is possible to adsorb and remove at least a portion of the inflammatory mediators contained in a liquid, thereby reducing their concentration. The liquid containing the inflammatory mediator is not particularly limited as long as it is one in which the concentration of the inflammatory mediator should be reduced, but examples thereof include biological fluids such as blood, lymph, plasma, serum, synovial fluid, and cerebrospinal fluid, with blood being preferred.

[0043] The biologically derived fluid in which the concentration of inflammatory mediators has been reduced by the inflammatory mediator adsorber of the present invention may be returned to the patient. Therefore, the inflammatory mediator adsorber of the present invention can also be incorporated into a hemodialysis machine for use, for example. [Example]

[0044] The present invention will be described in more detail below with reference to examples. However, the present invention is not limited to the following examples, and it is possible to carry out the invention by making appropriate modifications within the scope of the above and below-described aims, and all such modifications are included in the technical scope of the present invention.

[0045] Example 1: Preparation of dopamine-immobilized cellulose beads An alkaline aqueous solution was added to a suspension (100 mL) of porous cellulose beads (molecular weight exclusion limit: 40,000-60,000, particle size: 400-500 μm) to bring the total volume to 270 mL, and then epichlorohydrin (30 mL) was added and stirred at 40°C for 2 hours. After the reaction, the beads were thoroughly washed with water to obtain epoxidized cellulose beads. The resulting epoxidized cellulose beads were mixed with an aqueous dopamine solution, adjusted to a pH of 8.5 or higher with an alkaline solution, and then shaken at 60°C for 16 hours or more. After that, the beads were thoroughly washed with an acidic solution and water to obtain dopamine-immobilized cellulose beads. The amount of immobilized dopamine was measured by the Folin-Ciocalteu method and found to be 43 μmol / mL. Because dopamine was reacted under alkaline conditions, it is possible that at least a portion of it self-polymerized to coat the surface of the cellulose beads. However, the Folin-Ciocalteu method is a colorimetric method for quantifying tungsten molybdenum blue, a blue pigment formed by a redox reaction between phenolic hydroxyl groups and the Folin reagent, which is synthesized from sodium tungstate, sodium molybdate, and phosphoric acid. Therefore, it is possible to quantify reactive phenolic hydroxyl groups, and therefore active phenolic hydroxyl groups reactive with inflammatory mediators.

[0046] Example 2: Preparation of L-Dopa-immobilized cellulose beads L-DOPA-immobilized cellulose beads were obtained in the same manner as in Example 1, except that the dopamine aqueous solution was replaced with an L-DOPA aqueous solution. The amount of L-dopa immobilized was measured by the Folin-Ciocalteu method and found to be 30 μmol / mL.

[0047] Example 3: Preparation of 6-hydroxydopamine-immobilized cellulose beads 6-Hydroxydopamine-immobilized cellulose beads were obtained in the same manner as in Example 1, except that the dopamine aqueous solution was replaced with a 6-hydroxydopamine aqueous solution. The amount of immobilized 6-hydroxydopamine was measured by the Folin-Ciocalteu method and found to be 70 μmol / mL.

[0048] Example 4: Preparation of 6-hydroxy-L-dopa-immobilized cellulose beads 6-Hydroxy L-dopa-immobilized cellulose beads were obtained in the same manner as in Example 1, except that the dopamine aqueous solution was replaced with a 6-hydroxy L-dopa aqueous solution. The amount of immobilized 6-hydroxy-L-dopa was measured by the Folin-Ciocalteu method and found to be 35 μmol / mL.

[0049] Example 5: Preparation of L-dopa coated cellulose beads The L-DOPA solution was added to a suspension (100 mL) of porous cellulose beads (molecular weight exclusion limit: 40,000-60,000, particle size: 400-500 μm), and the pH was adjusted to 8.5 or higher with an alkaline solution. The mixture was then shaken at 60°C for 16 hours or more. The beads were then thoroughly washed with an acidic solution and water to obtain L-DOPA-coated cellulose beads. The amount of L-dopa immobilized was measured by the Folin-Ciocalteu method and found to be 20 μmol / mL. It is believed that the amount of active phenolic hydroxyl groups was slightly reduced compared to the L-DOPA-immobilized cellulose beads of Example 2, in which L-DOPA was covalently bonded via epoxy groups, because L-DOPA self-polymerized to coat the cellulose beads.

[0050] Comparative Example 1: Preparation of n-hexadecylamine-immobilized cellulose beads An aqueous ethanol solution containing n-hexadecylamine was added to the epoxidized gel produced in Example 1, and the mixture was allowed to react for 7 hours at 70° C. After the reaction was completed, the beads were thoroughly washed with ethanol and then water to obtain n-hexadecylamine-immobilized cellulose beads. The amount of immobilized n-hexadecylamine was measured by titration and found to be 17 μmol / mL.

[0051] Comparative Example 2: Preparation of phenylalanine-immobilized cellulose beads Phenylanine-immobilized cellulose beads were obtained in the same manner as in Example 1, except that the dopamine aqueous solution was replaced with a phenylalanine aqueous solution. The amount of immobilized phenylalanine was measured by elemental analysis and found to be 17 μmol / mL.

[0052] Comparative Example 3: Preparation of tyrosine-immobilized cellulose beads Tyrosine-immobilized cellulose beads were obtained in the same manner as in Example 1, except that the dopamine aqueous solution was replaced with a tyrosine aqueous solution. The amount of immobilized tyrosine was measured by the Folin-Ciocalteu method and found to be 14 μmol / mL.

[0053] Comparative Example 4: Preparation of glycine-immobilized cellulose beads Glycine-immobilized cellulose beads were obtained in the same manner as in Example 1, except that the dopamine aqueous solution was replaced with a glycine aqueous solution. The amount of immobilized glycine was measured by elemental analysis and found to be 13 μmol / mL.

[0054] Comparative Example 5: Preparation of diol-immobilized cellulose beads The amount of epoxy immobilized in the epoxidized gel produced in Example 1 was measured by titration. The epoxidized gel was sterilized by high-pressure steam in an autoclave at 120° C. for 1 hour to obtain diol-immobilized cellulose beads in which the epoxy groups had been ring-opened.

[0055] Test Example 1: Adsorption test of inflammatory mediators Inflammatory mediator solutions were prepared by dissolving bovine serum albumin (Sigma-Aldrich) at a concentration of 1% by mass and recombinant IL-6, TNFα, IL-10, IL-8, or calprotectin at the specified concentrations in phosphate buffer solution ("PBS(-)" Fujifilm Wako Pure Chemical Industries, Ltd.). Each inflammatory mediator solution (7.2 mL) was added to the centrifuge tube containing the adsorbent (0.4 mL) of Examples 1 to 5 and Comparative Examples 1 to 5, and the mixture was shaken for 2 hours at 37° C. Each inflammatory mediator solution (7.2 mL) was also added to a centrifuge tube containing no adsorbent, and the mixture was shaken in the same manner. After shaking, the concentrations of each inflammatory mediator in the supernatant of each centrifuge tube were measured by ELISA. The removal rate (%) of each inflammatory mediator by the adsorbent was calculated from the difference between the supernatant concentrations in the centrifuge tubes without adsorbent and those with adsorbent. The removal rates (%) of each inflammatory mediator are shown in Table 1. The concentrations of IL-6, TNFα, IL-10, IL-8, calprotectin, and HMGB1 in the centrifuge tubes without adsorbent after shaking were 3906 pg / mL, 1028 pg / mL, 857 pg / mL, 137 pg / mL, and 75 ng / mL, respectively.

[0056] [Table 1]

[0057] As shown in Table 1, the adsorption performance of inflammatory mediators was not sufficient when only an amine compound was attached as a ligand. Even when phenylalanine, which has only one phenolic hydroxyl group on the phenyl ring, was attached, the adsorption performance of inflammatory mediators was not at all sufficient. In contrast, when a ligand having two phenolic hydroxyl groups on the phenyl ring was attached, inflammatory mediators were successfully adsorbed. Thus, it was demonstrated that the adsorbent according to the present invention can efficiently adsorb and remove inflammatory mediators. [Explanation of symbols]

[0058] 1a: Inlet, 1b: Lid, 2a: Outlet, 2b: Lid, 3: Adsorbent, 4: Filter, 5: Filter, 6: Housing

Claims

1. a water-insoluble carrier and a ligand for an inflammatory mediator; the ligand has an aromatic hydrocarbon group having two or more phenolic hydroxyl groups, An inflammatory mediator adsorbent, characterized in that the ligand is bound to the water-insoluble carrier.

2. 2. The inflammatory mediator adsorbent according to claim 1, wherein at least two of said phenolic hydroxyl groups are adjacent to each other on said aromatic hydrocarbon group.

3. 2. The inflammatory mediator adsorbent according to claim 1, wherein the aromatic hydrocarbon group is a catechol group.

4. The ligand is attached to the water-insoluble carrier by -N(R 1 )-group (wherein, R 1 is H or C 1-6 2. The inflammatory mediator adsorbent according to claim 1, wherein the inflammatory mediator is covalently bonded via a group (representing an alkyl group).

5. The inflammatory mediator adsorbent according to claim 1, wherein the inflammatory mediator is one or more inflammatory mediators selected from IL-6, TNFα, IL-10, IL-8, calprotectin, and HMGB1.

6. 2. The inflammatory mediator adsorbent according to claim 1, wherein the water-insoluble carrier comprises a cellulose derivative.

7. a housing; the housing having a liquid inlet and an outlet; An inflammatory mediator adsorbent, characterized in that the housing is filled with the inflammatory mediator adsorbent according to any one of claims 1 to 6.

8. A method for producing an inflammatory mediator adsorbent, comprising: A method comprising the step of covalently bonding a ligand to a water-insoluble support by reacting a ligand precursor compound having an aromatic hydrocarbon group having two or more phenolic hydroxyl groups with the support.

9. The method of claim 8, wherein the ligand precursor compound is L-3,4-dihydroxyphenylalanine or dopamine.

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