Water-insoluble carrier for removal of activated platelets

Abstract

The purpose of the present invention is to provide an insoluble carrier for removing activated platelets. The present invention provides a water-insoluble carrier which has a surface kurtosis of 0.1 to 16.0 and which removes activated platelets.

Description

Technical Field

[0001] The present invention relates to a water-insoluble carrier for removing activated platelets. Background Art

[0002] Thrombosis is involved in the development of morbid conditions such as arteriosclerosis, cancer, infection or renal disease, and activated platelets are involved in the formation of thrombus. Therefore, in order to suppress the formation of thrombus, attempts have been made to treat by giving antiplatelet drugs, but antiplatelet drugs not only suppress the function of activated platelets, but also suppress the function of non-activated platelets, so there is concern about bleeding in the skin, nosebleeds or gums as side effects. Wherein, in the case of severe bleeding, platelet preparations must sometimes be transfused, so the method for selectively removing activated platelets is expected. In addition, the growth factors released by activated platelets are also involved in the development of morbid conditions such as arteriosclerosis, cancer, infection or renal disease. Wherein, it is also known that platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF) are involved in the tumor growth of cancer, and vascular endothelial growth factor (VEGF) is involved in the development of infection etc. Therefore, removing these growth factors, the method for suppressing morbid development is also expected.

[0003] Patent Document 1 discloses a material that can be removed by phagocytosis of activated leukocyte-activated platelet complexes by setting the surface development length ratio of a water-insoluble carrier to which a compound containing a charged functional group is bound to the surface to 4 to 7. This material can be used to treat inflammatory diseases such as respiratory diseases.

[0004] Patent Document 2 discloses that a protein adsorbing material in which the generation of fine particles is suppressed can be used for adsorbing latent transforming growth factor-β by specifying the cross-sectional shape of the sea-island composite fiber.

[0005] Patent Document 3 discloses an adsorbent material capable of adsorbing immunosuppressive proteins by specifying the amino group amount and carbon atom number of the aliphatic primary and secondary amines, and for use in cancer treatment for inhibiting platelet adsorption.

[0006] Patent Document 4 discloses an adsorbent material capable of adsorbing immunosuppressive leukocytes by specifying the structure, number of carbon atoms, and arithmetic mean roughness of a polyamine, and for use in cancer treatment by inhibiting platelet adsorption.

[0007] Prior art literature

[0008] Patent Literature

[0009] Patent Document 1: International Publication No. 2018 / 225764

[0010] Patent Document 2: International Publication No. 2019 / 045031

[0011] Patent Document 3: International Publication No. 2019 / 049961

[0012] Patent Document 4: International Publication No. 2019 / 049962 Summary of the Invention

[0013] Problems to be solved by the invention

[0014] However, the adsorbent described in Patent Document 1 targets the activated leukocyte-activated platelet complex for removal, and does not mention at all a technique for removing activated platelets alone.

[0015] The adsorbent described in Patent Document 2 is a protein adsorbent that suppresses the generation of microparticles, and does not mention at all a technique for removing activated platelets.

[0016] The purpose of the adsorption materials described in Patent Documents 3 or 4 is to adsorb immunosuppressive proteins or immunosuppressive leukocytes. In contrast to the present application, they are materials that inhibit platelet adsorption and do not mention any technology related to the removal of activated platelets.

[0017] Therefore, an object of the present invention is to provide an insoluble carrier for removing activated platelets.

[0018] Means for solving problems

[0019] The configuration of the present invention for solving the above-mentioned problems is as follows.

[0020] (1) A water-insoluble carrier having a surface kurtosis of 0.1 to 16 and capable of removing activated platelets.

[0021] (2) The water-insoluble carrier according to (1) above, wherein the surface has an electric charge.

[0022] (3) The water-insoluble carrier according to (1) or (2) above, wherein the surface charge is 0.3 to 3.0 mmol per dry mass 1 g.

[0023] (4) The water-insoluble carrier according to any one of (1) to (3) above, wherein the surface has an amino group.

[0024] (5) The water-insoluble carrier according to any one of (1) to (4) above, which is a fiber or a particle.

[0025] (6) The water-insoluble carrier according to (5) above, which has a diameter of 1 to 100 μm.

[0026] (7) The water-insoluble carrier according to any one of (1) to (6) above, comprising a polymer selected from the group consisting of polystyrene, polypropylene, polysulfone, polyethersulfone and cellulose.

[0027] (8) The water-insoluble carrier according to any one of (1) to (7) above, wherein platelet-derived growth factor, vascular endothelial growth factor, and / or fibroblast growth factor are removed.

[0028] (9) A blood purifier comprising the water-insoluble carrier according to any one of (1) to (8) above.

[0029] (10) The blood purifier according to (9) above, which is a blood purification column.

[0030] (11) The blood purifier according to (9) or (10) above, which is used to inhibit thrombosis.

[0031] Effects of the Invention

[0032] According to the water-insoluble carrier, blood purifier and blood purification column of the present invention, activated platelets can be removed efficiently. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 : is a schematic diagram showing an example of an image used for kurtosis analysis.

[0034] Figure 2 : Schematic diagram showing the platelet fraction among CD41-positive cells in flow cytometry. DETAILED DESCRIPTION

[0035] The present invention is a water-insoluble carrier having a surface kurtosis of 0.5 to 16.0 and capable of removing activated platelets.

[0036] A "water-insoluble carrier" refers to a carrier that is insoluble in water. In this context, "water-insoluble" means that the change in dry mass of the carrier before and after immersion in water is 1% by mass or less. This change in dry mass is the ratio of the dry mass of the solid component remaining after immersing the carrier in 9 times the dry mass of the carrier at 37°C water for 1 hour, lifting the carrier with tweezers, etc., and vacuum drying the remaining water at 50°C or below to a constant weight, relative to the dry mass of the carrier before immersion. If the carrier is not water-insoluble, there is a risk of increased elution during actual use, which is not preferred from a safety perspective.

[0037] The material of the water-insoluble carrier of the present invention is not particularly limited as long as it can maintain the strength of the water-insoluble carrier. From the perspective of maintaining strength and easiness of processing the surface shape of the water-insoluble carrier, for example, synthetic polymers such as polystyrene, polypropylene, polyethylene, polysulfone, polyethersulfone, polyester, nylon, or polyvinyl alcohol, or natural polymers such as cellulose, collagen, chitin, chitosan, or dextran, or modified natural polymers such as cellulose esters such as cellulose triacetate can be mentioned. These polymers can be homopolymers or copolymers, or two or more polymers can be blended or alloyed for use.

[0038] Especially when used for blood purification, from the perspective of proven biocompatibility, it is preferred to include one or more polymers selected from polystyrene, polypropylene, polysulfone, polyethersulfone, and cellulose. Among them, polystyrene is particularly preferred due to its high number of aromatic rings per unit mass and ease of introduction of various functional groups and reactive functional groups via Friedel-Crafts reactions. It should be noted that the polymers contained in these water-insoluble carriers can be commercially available products or those produced by known methods.

[0039] The surface of the water-insoluble carrier of the present invention, when kurtosis is used as an indicator, refers to the region of the water-insoluble carrier where surface irregularities can be detected using a laser microscope in accordance with JIS B 0601:2001. When charge is used as an indicator, it refers to the region of the water-insoluble carrier where charged functional groups or compounds containing charged functional groups are bound. For example, when the water-insoluble carrier is porous, the surface layer within each pore is also included in the surface.

[0040] The shape of the water-insoluble carrier of the present invention can be, for example, a film, a particle or a fiber. Among them, from the viewpoint of easily ensuring the blood flow path, fibers or particles are preferred. Furthermore, in the case of being used for blood purification purposes, fibers with a large specific surface area, flexible deformation and excellent operability are more preferred. From the viewpoint of maintaining strength and being easy to process into a blood purifier, sea-island composite fibers are further preferred. In addition, from the viewpoint of ease of filling the water-insoluble carrier during use and that the liquid flow path can be uniform, the fiber is preferably processed into a form such as a non-woven fabric, a knitted fabric or a fabric for use.

[0041] The water-insoluble carrier for removing activated platelets of the present invention may be a water-insoluble carrier alone or a carrier obtained by fixing or mixing a suitable reinforcing material onto the water-insoluble carrier. The fixing or mixing operation may be performed before or after processing into an adsorption carrier such as a blood purifier.

[0042] When the water-insoluble carrier of the present invention is in the form of fibers or particles, the diameter of the carrier is preferably 1 μm or greater. From the perspective of ensuring a flow path through which blood cells can pass, it is more preferably 3 μm or greater, and even more preferably 5 μm or greater. Furthermore, from the perspective of ensuring a specific surface area for adsorption, it is preferably 100 μm or less, more preferably 60 μm or less, and even more preferably 40 μm or less. In summary, the diameter of the water-insoluble carrier is preferably 1 to 100 μm, more preferably 3 to 60 μm, and even more preferably 5 to 40 μm. Any preferred lower limit value may be combined with any preferred upper limit value.

[0043] "Kurtosis" (hereinafter referred to as "Rku") is an indicator that quantifies the sharpness of the height distribution defined in JIS B 0601:2001. It is a parameter that allows the sharpness of the surface shape of a water-insoluble carrier to be determined based on its numerical value. Rku can be calculated by taking an image of the surface of the water-insoluble carrier using a laser microscope and analyzing the image using analysis software. To account for surface variations in the water-insoluble carrier, it is appropriate to analyze the image extracted from the surface.

[0044] like Figure 1 As shown, Rku represents the fourth-power average of Z(x) in the reference length obtained by dimensionlessly converting the root mean square height (Zq) of the extracted portion to the fourth power, extracted along a right angle direction with a reference length l in an image of the surface of the water-insoluble carrier obtained using a laser microscope. It can be calculated from an image obtained using a laser microscope (e.g., Ultra-Deep Color 3D Profiling Microscope VK-9710, manufactured by KEYENCE Corporation) using a surface roughness analysis function (VKViewer ver. 2.5 and VKAnalyzer ver. 2.5, manufactured by KEYENCE Corporation) using the following formula (1). Specifically, the measurement is performed by the "Measurement of Kurtosis" described later.

[0045] [Number 1]

[0046]

[0047] The reference length l extracted from the surface image of the water-insoluble carrier is set to 5 to 100 μm. If the shortest side of the water-insoluble carrier in the extracted image is less than 5 μm or greater than 200 μm, the reference length l is set to 1 / 5 to 1 / 2 of the length of the shortest side. If the water-insoluble carrier is a fiber, the diameter calculated from the area of ​​the cross section perpendicular to the fiber's elongation direction can be used as the length of the shortest side. In the case of particles, the diameter of a circle having the same area as the particle's cross-sectional area in the extracted image can be used.

[0048] In the case of removing activated platelets, when the height distribution of the surface of the water-insoluble carrier is in a shape that is biased toward the upper part (mountain) relative to the average surface, that is, when the surface has a gentle concave-convex shape, the activated platelets extend pseudopodia (or false feet), thereby promoting aggregation and adhesion on the material and improving the removal performance. On the other hand, when the height distribution is in a shape that is protruding into a needle-like shape relative to the average surface, that is, when the surface has a sharp concave-convex shape, the aggregation and adhesion of the activated platelets are suppressed and the removal performance is reduced. Therefore, the Rku of the surface needs to be 0.1 to 16.0, preferably 0.1 to 13.0, more preferably 0.1 to 10.0, further preferably 0.1 to 6.0, and particularly preferably 0.1 to 3.0. Any preferred lower limit value can be combined with any preferred upper limit value.

[0049] Rku can be controlled by, for example, adjusting the temperature or time of immersing the water-insoluble carrier in a solution capable of dissolving the water-insoluble carrier, adjusting the concentration of the crosslinking agent that inhibits dissolution of the water-insoluble carrier, or adjusting the temperature or time of reacting the water-insoluble carrier with the crosslinking agent.

[0050] When the water-insoluble carrier is a polymer having an aromatic ring such as polystyrene, for example, paraformaldehyde (hereinafter referred to as "PFA") can be used as the cross-linking agent.

[0051] The water-insoluble carrier of the present invention preferably has an electric charge on its surface.

[0052] "Surface charge" refers to the presence of positively or negatively charged functional groups or compounds containing positively or negatively charged functional groups on the surface of the water-insoluble carrier. The compound containing positively or negatively charged functional groups is not limited as long as it can interact with the target substance, such as a growth factor. Examples of chemical structures include compounds containing positively charged functional groups (cationic functional groups), such as amino groups, or compounds containing negatively charged functional groups (anionic functional groups), such as sulfonic acid groups or carboxyl groups. It should be noted that these functional groups may be a combination of multiple identical or different functional groups.

[0053] It should be noted that compounds containing a charged functional group may also contain uncharged functional groups as long as they contain the above-mentioned charged functional group. For example, compounds in which an aryl group such as an alkyl group such as a methyl group or an ethyl group, a phenyl group, or a phenyl group substituted with an alkyl group (e.g., p-(p)-methylphenyl, m-(m)-methylphenyl, o-(o)-methylphenyl, p-(p)-ethylphenyl, m-(m)-ethylphenyl, or o-(o)-ethylphenyl), or a phenyl group substituted with a halogen atom (e.g., p-(p)-fluorophenyl, m-(m)-fluorophenyl, o-(o)-fluorophenyl, p-(p)-chlorophenyl, m-(p)-chlorophenyl, or o-(o)-chlorophenyl) is bonded to a charged functional group are also included in compounds containing a charged functional group.

[0054] From the perspective of achieving sufficient interaction with growth factors and ensuring the degree of freedom of the steric configuration of the introduced functional groups while achieving appropriate interaction with the removal target substance, the surface charge of the water-insoluble carrier of the present invention is preferably 0.3 to 3.0 mmol, more preferably 0.5 to 2.0 mmol, and even more preferably 0.5 to 1.5 mmol per 1 g of the dry mass of the water-insoluble carrier. Any preferred lower limit value may be combined with any preferred upper limit value.

[0055] The charge amount on the surface of the water-insoluble carrier can be controlled by, for example, adjusting the type of functional group introduced into the water-insoluble carrier or the concentration of the solution of the compound containing a functional group that is charged during the reaction, or the reaction time.

[0056] From the viewpoint of excellent biocompatibility, the water-insoluble carrier of the present invention preferably has an amino group on the surface. Examples of compounds having an amino group include compounds having an amino group derived from a primary amine such as methylamine, ethylamine, propylamine, butylamine, pentylamine, hexylamine, heptylamine, octylamine or dodecylamine, compounds having an amino group derived from a secondary amine such as methylhexylamine, diphenylmethylamine or dimethylamine, amino groups derived from amines having an unsaturated alkyl chain such as allylamine, compounds having an amino group derived from a tertiary amine such as trimethylamine, triethylamine, dimethylethylamine, phenyldimethylamine or dimethylhexylamine, 1-(3-aminopropyl)imidazole, pyridine-2-yl pyridine, 1-hydroxy-1-methyl-2-imidazol ... -A compound having an amino group derived from an amine having an aromatic ring, such as tris(2-aminoethyl)amine, ethylenediamine, diethylenetriamine, triethylenetetramine, tetraethylenepentamine (hereinafter referred to as "TEPA"), dipropylenetriamine, polyethyleneimine, N-methyl-2,2'-diaminodiethylamine, N-acetylethylenediamine or 1,2-bis(2-aminoethoxyethane) (hereinafter referred to as "polyamine"), to which two or more amino groups are bonded via an alkyl chain, an aromatic compound, a heterocyclic compound or a monocyclic compound. Among them, from the viewpoint of being able to efficiently adsorb growth factors released by activated platelets, the amino group possessed by the water-insoluble carrier on the surface is preferably an amino group derived from a polyamine. In addition, from the viewpoint of having a high degree of freedom in the stereo configuration of the functional group and being able to efficiently adsorb growth factors released by activated platelets, the amino group is more preferably an amino group derived from a primary amine or a secondary amine.

[0057] The compound having a sulfonic acid group may be any compound having at least one sulfonic acid group, and examples thereof include aliphatic sulfonic acids such as sulfonic acid and methanesulfonic acid, aromatic sulfonic acids such as benzenesulfonic acid, p-(p)-phenolsulfonic acid, and 4-methylbenzenesulfonic acid, and halogenated sulfonic acids such as fluorosulfonic acid and chlorosulfonic acid.

[0058] The compound having a carboxyl group may be any compound having at least one carboxyl group, and examples thereof include aliphatic carboxyl groups such as acetic acid and propionic acid, and aromatic carboxyl groups such as phenylcarboxyl groups.

[0059] The water-insoluble carrier and the compound containing a charged functional group may be directly bonded, or a spacer derived from a reactive functional group may be interposed between the water-insoluble carrier and the compound containing a charged functional group. The spacer may be any spacer having an electrically neutral chemical bond such as an amide bond, a urea bond, an ether bond, or an ester bond, with spacers having an amide bond or a urea bond being preferred.

[0060] Examples of the reactive functional group that mediates the bond between the water-insoluble carrier and the compound containing a charged functional group include active halogen groups such as a halomethyl group, a haloacetyl group, a haloacetamidomethyl group, or a haloalkyl group, an epoxide group, a carboxyl group, an isocyanate group, a thioisocyanate group, or an acid anhydride group. Among these, active halogen groups are preferred, and haloacetamidomethyl groups are more preferred, from the perspective of having moderate reactivity.

[0061] The reactive functional group can be combined with the water-insoluble carrier by reacting the water-insoluble carrier with an appropriate reagent in advance. For example, when the water-insoluble carrier is polystyrene and the reactive functional group is a chloroacetamidomethyl group, a polystyrene bound to a chloroacetamidomethyl group can be obtained by reacting the polystyrene with N-hydroxymethyl-2-chloroacetamide (hereinafter referred to as "NMCA"). By reacting the polystyrene bound to a chloroacetamidomethyl group with, for example, TEPA having an amino group, a polystyrene to which TEPA is bound via an acetamidomethyl group can be obtained. In this case, TEPA is a compound containing a functional group with a charge. It should be noted that the starting materials and reagents used in the manufacture of the water-insoluble carrier for removing activated platelets can use commercially available products or those manufactured by known methods.

[0062] Since the compound containing a charged functional group needs to interact with the substance to be removed in the blood, it is preferably bound to at least the surface of the water-insoluble carrier that comes into contact with the blood.

[0063] The charge of a water-insoluble carrier can be measured by acid-base titration using hydrochloric acid or sodium hydroxide aqueous solution. Specifically, it can be measured using the methods described below in "Determination of Positive Charge" or "Determination of Negative Charge." Both positive and negative charges are expressed as absolute values.

[0064] The "fiber diameter" can be determined by the following method. First, randomly sample 100 dry fiber samples and, using a scanning electron microscope or the like, take a photograph of a cross section perpendicular to the fiber's direction of extension for each sample at a magnification of 1000 to 3000. Next, measure the diameter of each fiber cross section and calculate the average of these values ​​(the average of the diameters of all 100 fiber cross sections) to determine the "fiber diameter." If the fiber cross section is not circular, the diameter of the fiber is the diameter of a circle having the same area as the cross section.

[0065] The "diameter of the particles" can be determined by the following method. First, 10 samples of dried particles are randomly collected and a photograph of each sample is taken at a magnification of 1000 to 3000 using a scanning electron microscope or the like. Next, the diameters of 10 particles are measured for each photograph, and the average of these values ​​(the average of the diameters of a total of 100 particles) is calculated to determine the "diameter of the particles." If the shape of the particles photographed is not circular, the diameter of the particle is the diameter of a circle having the same area as the area of ​​the particles in the photograph.

[0066] "Blood" means a liquid containing proteins, lipids, blood cell components, etc. Specifically, it includes buffer solutions containing proteins, lipids, blood cell components, etc., body fluids, blood, plasma, serum, etc.

[0067] "Blood components" refer to components that constitute blood. Examples include blood cell components such as red blood cells, white blood cells, and platelets, and fluid factors such as growth factors released from these cells. When the goal is to inhibit thrombosis, it is preferable to remove growth factors from fluid factors in addition to activated platelets.

[0068] “Activated platelets” refer to platelets that have the function of extending amoeba-like projections called pseudopodia (or pseudopodia) and aggregating to form thrombi due to reactions such as bleeding, inflammation, immunity, infection defense, arteriosclerosis, or cancer metastasis.

[0069] The degree of platelet activation can be determined by measuring the expression of platelet surface antigens using flow cytometry or the like. Quantitative measurement of platelet surface antigen expression using flow cytometry or the like is preferred from the perspective of quantitatively evaluating platelet activation over morphologically subjective determination of platelet pseudopodia elongation, deformation, adhesion, or aggregation.

[0070] In addition, known platelet surface antigens include CD41, CD42, CD42b, CD61, and CD62P. From the perspective of detecting activated platelets, a method for detecting CD62P, which is specifically expressed on the platelet membrane, is preferred. Furthermore, activated platelets may bind to red blood cells or white blood cells, but the term "activated platelets" as used herein refers to activated platelets that are not bound to red blood cells or white blood cells.

[0071] "Non-activated platelets" refer to platelets in which CD62P cannot be detected using flow cytometry or other methods, or platelets that do not extend amoebic protrusions called pseudopodia (or pseudopods) and deform, adhere, or aggregate, thus lacking the ability to form thrombi. The degree of inactivation can be determined by measuring the expression of platelet surface antigens using flow cytometry or other methods.

[0072] The activated platelet concentration can be calculated, for example, by reacting a platelet fraction from peripheral blood with an activation detection reagent (activated platelet binding reagent) that specifically binds to activated platelets, and measuring the fraction in the platelet fraction from peripheral blood that specifically binds to the activated platelet binding reagent.

[0073] The activated platelet detection reagent does not bind to non-activated platelets but has binding properties to activated platelets. For example, activated platelets can be detected by using an anti-CD62P antibody (e.g., anti-human CD62P (P-selectin) antibody, BioLegend.) that detects CD62P, which is known to be a specific cell surface marker for activated platelets.

[0074] The water-insoluble carrier of the present invention is characterized in that it removes activated platelets. Removal of activated platelets means that the activated platelet removal rate [A] of the water-insoluble carrier is divided by the non-activated platelet removal rate [B] of the water-insoluble carrier, that is, the activated platelet removal ratio is greater than 1.5 ([A] / [B]>1.5). The larger the activated platelet removal ratio, the more selectively activated platelets are removed. Therefore, the activated platelet removal ratio is preferably 2.0 or greater, more preferably 3.0 or greater, and even more preferably 5.0 or greater.

[0075] From the viewpoint of inhibiting thrombosis, the activated platelet removal rate is preferably 20% or more, more preferably 30% or more, further preferably 40% or more, and particularly preferably 60% or more.

[0076] "Growth factor" means a molecule with physiological activity that promotes the growth of cells or tissues. Useful growth factors in the present invention include: platelet-derived growth factor (hereinafter referred to as "PDGF"), vascular endothelial growth factor (hereinafter referred to as "VEGF"), fibroblast growth factor (hereinafter referred to as "FGF") and mixtures of the above factors. PDGF includes PDGF-AA, PDGF-BB, PDGF-AB, PDGF-CC and PDGF-DD. VEGF includes VEGF-A, VEGF-B, VEGF-C, VEGF-D and VEGF-E. In addition, FGF includes FGF1 to 23. In addition, the above growth factors can also be combined with other proteins.

[0077] The water-insoluble carrier of the present invention preferably has PDGF, VEGF, and / or FGF removed. By removing activated platelets and simultaneously removing PDGF, VEGF, and / or FGF, not only can thrombosis be inhibited, but the progression of conditions in which PDGF, VEGF, and / or FGF are involved can also be inhibited. Examples of conditions in which PDGF, VEGF, and / or FGF are involved include arteriosclerosis, cancer, inflammatory diseases, infections, and renal disease. From the perspective of inhibiting the progression of these conditions, the PDGF, VEGF, and / or FGF removal rate is preferably 20% or greater, more preferably 50% or greater, further preferably 70% or greater, and particularly preferably 80% or greater.

[0078] "Blood purifier" means a device that removes substances from blood to purify the blood.

[0079] The blood purifier of the present invention is characterized by comprising the water-insoluble carrier of the present invention. As long as it can purify blood, the shape of the device is not limited, and a blood purification column, a blood purification filter, a blood purification bag, a blood purification unit, a blood purification housing, or a blood purification syringe can be suitably used.

[0080] The “blood purification column” refers to a blood purifier having at least a blood inlet, a blood outlet, and a shell, and a water-insoluble carrier inside the shell.

[0081] The blood purification column of the present invention is characterized by comprising the water-insoluble carrier of the present invention. As the shape of the blood purification column, for example, a radial flow column is preferably used.

[0082] A "thrombus" is a clot formed by the aggregation and coagulation of platelets in the blood. When blood vessels are narrowed by a thrombus and blood circulation is reduced, it can cause circulatory problems and organ dysfunction. Furthermore, if blood is not flowing, nutrients cannot reach the organs ahead, leading to cell death and dysfunction.

[0083] The blood purifier comprising the water-insoluble carrier of the present invention can be suitably used for inhibiting thrombosis. The application is not particularly limited as long as it is for inhibiting thrombosis, and can be used for diseases caused by thrombosis, for example, venous thrombosis in which thrombosis occurs in veins or arterial thrombosis in which thrombosis occurs in arteries.

[0084] Examples of diseases for which therapeutic effects can be expected by inhibiting thrombosis include atherosclerosis, myocardial infarction, ischemic heart disease, cerebral infarction, stroke, valvular heart disease, pulmonary embolism, angina pectoris, dementia, hyperlipidemia, dyslipidemia, diabetes, economy class syndrome, arterial occlusive disease, deep vein thrombosis, portal vein thrombosis, intermittent claudication, quadriplegia, hearing impairment, pain, sepsis, disseminated intravascular coagulation (DIC), bacteremia, viral infection, toxin infection, COVID-19 infection, acute lung injury (ALI), acute respiratory distress syndrome (also known as ARDS, acute respiratory distress syndrome, acute respiratory distress syndrome), pneumonia, acute respiratory insufficiency, septic shock, toxic shock syndrome, multiple organ failure, chronic obstructive pulmonary disease, Kawasaki disease, and cancer.

[0085] Example

[0086] The present invention is described below in detail using experimental examples and comparative examples, but the present invention is not limited to these examples. First, the measurement and evaluation methods are described below. As an example, the measurement method is described for a case where the water-insoluble carrier is a knitted fabric, but the same method can also be used for measurements in forms other than knitted fabrics.

[0087] (Determination of Kurtosis)

[0088] Take out about 0.3 g of the water-insoluble carrier and vacuum dry it at 15 to 40°C for more than 8 hours. Use double-sided tape to fix the vacuum-dried water-insoluble carrier on the observation platform of a laser microscope (ultra-deep color 3D shape measurement microscope VK-9710, manufactured by KEYENCE Corporation). The objective lens uses a long-distance lens (model: EPIPLAN / ELWD100, manufactured by KEYENCE Corporation), sets the magnification to 100 times, the optical zoom to 1 times, and does not use a neutral density filter. Use a laser microscope to take a picture of the surface of the water-insoluble carrier, and use the surface roughness analysis function (VKViewerver.2.5 and VKAnalyzerver.2.5, manufactured by KEYENCE Corporation) on the obtained image to calculate Rku by the following formula (1).

[0089] Here, Zq means root mean square height, which is expressed by the following formula (2): Analysis of images captured by a laser microscope for calculating Rku is performed as follows because the obtained value may vary depending on the conditions.

[0090] The analysis was conducted using the method described in JIS B 0601:2001, employing surface roughness without moiré correction. Images collected at one location constituted 10 fields of view, and images collected at three locations were analyzed for a total of 30 fields of view. The reference length l was set to 10 μm vertically and 10 μm horizontally, and the extracted area was analyzed as one field of view.

[0091] Taking into account surface variations, the Rku value of each sample of the water-insoluble carrier was calculated as the average value of 30 fields of view. However, if the surface shape of the water-insoluble carrier varies, appropriate evaluation cannot be performed, so the average value was calculated excluding the outliers.

[0092] The interquartile range (IQR) was used to determine the outliers. The quartiles were calculated from the Rku of 30 fields of view, and the outliers were set as "values ​​greater than the value obtained by adding 1.5 times the interquartile range to the 3rd quartile" and "values ​​less than the value obtained by subtracting 1.5 times the interquartile range from the 1st quartile" and excluded from the calculation of the average value. It should be noted that when there are more than 7 fields of view as outliers, the same analysis is performed again starting from the image read into the laser microscope. It should be noted that the value of Rku is rounded to the second decimal place.

[0093] [Number 2]

[0094]

[0095] [Number 3]

[0096]

[0097] (Determination of positive charge)

[0098] The cationic charge of the water-insoluble carrier is calculated by acid-base back titration of the cationic functional groups in the water-insoluble carrier.

[0099] Use a punching machine (diameter: 28mm) to punch out 6 pieces of knitted fabric as a water-insoluble carrier. Add the punched water-insoluble carrier to a polypropylene centrifuge tube (50mL). Then, add a 6mol / L sodium hydroxide aqueous solution to the extent of immersion of the water-insoluble carrier, and stir it upside down with a rotator for more than 30 minutes (desalination treatment of positive charge). Discard the 6mol / L sodium hydroxide aqueous solution from the centrifuge tube, add ion exchange water to the extent of immersion of the water-insoluble carrier, stir it with a vortex for more than 1 second, and discard the cleaning solution. Repeat this cleaning operation more than 5 times, and use phenolphthalein solution to confirm that the pH of the final cleaning solution is neutral.

[0100] Next, the water-insoluble carrier was placed in a vacuum dryer and dried at 15-40°C. The vacuum drying endpoint was set at least 8 hours after drying. Vacuum drying was terminated when the mass reduction rate of the second value measured using an electronic balance at least 15 minutes after the first measurement was less than 5%. If the mass reduction rate was 5% or more, vacuum drying was continued for an additional 15 minutes or more until the mass reduction rate was less than 5%. The same procedure was repeated to determine the dry mass [a] of the water-insoluble carrier.

[0101] Next, the vacuum-dried water-insoluble carrier was placed in a polypropylene centrifuge tube (50 mL), 25 mL of 0.1 mol / L hydrochloric acid [b] was added, and the mixture was stirred inverted on a rotator for 30 minutes. The stirred solution [c] was dispensed into centrifuge tubes in 5 mL increments, and 30 μL of methyl red solution and phenolphthalein solution were added to prepare the measurement solution. Three tubes of measurement solution were prepared.

[0102] Next, titrate with a 0.05 mol / L sodium hydroxide aqueous solution [d]. The endpoint is the time when the test solution turns yellow after being inverted and mixed three times. The three tubes of test solution are titrated, and the average amount of sodium hydroxide aqueous solution added used in the titration is set as the average amount added [e]. The positive charge is calculated using the following formula (3).

[0103] Positive charge per dry mass 1g (mmol / g) = (amount of dispensed solution of [c] × concentration of [b] / concentration of [d] - [e]) × concentration of [d] × (amount added of [b] / amount of dispensed solution of [c]) / [a] ···Formula (3)

[0104] In addition, the positive charge amount is a value obtained by rounding off to the second decimal place.

[0105] (Determination of negative charge)

[0106] The negative charge of the water-insoluble carrier is calculated by acid-base back titration of the anionic functional groups in the water-insoluble carrier.

[0107] Use a punching machine (diameter: 28mm) to punch out 6 pieces of knitted fabric serving as a water-insoluble carrier. Add the punched water-insoluble carrier to a polypropylene centrifuge tube (50mL). Then, add 6mol / L hydrochloric acid to the extent of immersing the water-insoluble carrier, and stir with a rotator for more than 30 minutes (negative charge desalination treatment). Discard the 6mol / L hydrochloric acid from the centrifuge tube, add ion exchange water to the extent of immersing the water-insoluble carrier, stir with a vortex for more than 1 second, and discard the cleaning solution. Repeat this cleaning operation more than 5 times, and use a methyl red solution to confirm that the pH of the final cleaning solution is neutral.

[0108] Next, the water-insoluble carrier was placed in a vacuum dryer and dried at 15-40°C. The endpoint of vacuum drying was set at least 8 hours after drying. Vacuum drying was terminated when the mass reduction rate of the second value measured using an electronic balance at least 15 minutes after the first measurement was less than 5%. If the mass reduction rate was 5% or greater, vacuum drying was continued for an additional 15 minutes or more until the mass reduction rate was less than 5%. The same procedure was repeated to determine the dry mass [f] of the water-insoluble carrier.

[0109] Next, the vacuum-dried water-insoluble carrier was placed in a polypropylene centrifuge tube (50 mL), 25 mL of a 0.1 mol / L sodium hydroxide aqueous solution [g] was added, and the mixture was stirred inverted for 30 minutes using a rotator. 5 mL of the stirred solution [h] was dispensed into the centrifuge tube, and 5 mL of 0.1 mol / L hydrochloric acid [i] was added. 30 μL of methyl red solution and phenolphthalein solution were added to prepare the measurement solution. 3 tubes of measurement solution were prepared.

[0110] Next, titrate with a 0.05 mol / L sodium hydroxide aqueous solution [j]. The endpoint is the time when the test solution turns yellow after inverting and mixing three times. The three tubes of test solution are titrated, and the average amount of sodium hydroxide aqueous solution added used in the titration is set as the average amount of droplets [k]. The negative charge is calculated using the following formula (4).

[0111] Negative charge per dry mass 1g (mmol / g) = [k] × [j] concentration × ([g] added amount / [h] dispensed amount) / [f] Formula (4)

[0112] In addition, the negative charge amount is a value rounded off to the second decimal place.

[0113] (Determination of fiber diameter)

[0114] 100 fibers were randomly collected from a knitted fabric serving as a dry, water-insoluble carrier. A scanning electron microscope was used to photograph a cross-section perpendicular to the fiber's direction of extension at 2000x magnification for each sample. Next, the cross-sectional area of ​​each fiber was determined using analysis software (Photoshop Elements 14.0, manufactured by Adobe Systems). The diameter of a circle having the same area as the obtained cross-sectional area was calculated. The average of these values ​​(the average of the diameters of all 100 fiber cross-sections) was defined as the fiber diameter. Note that the fiber diameter was rounded off to the first decimal place.

[0115] (Determination of removal rate of activated platelets and non-activated platelets)

[0116] The water-insoluble carriers cut into 1 cm diameter disks were stacked and filled into a cylindrical column (1 cm inner diameter × 1.2 cm height, 0.94 cm inner volume) with a solution inlet and outlet. 3 , outer diameter 2 cm, made of polycarbonate), thereby preparing a column.

[0117] Heparin was added to the blood of healthy volunteers to achieve a concentration of 5 units / mL. Adenosine diphosphate (ADP) was then added to the blood of healthy volunteers to achieve a concentration of 2 μmol / L. The blood was then activated by shaking in a hot water bath at 37°C for 30 minutes at 65 rpm.

[0118] Next, activated blood was pumped through the column at a flow rate of 0.63 mL / min, and blood was collected at the column inlet and outlet. The column inlet sample was collected from blood immediately before the column was immersed in a hot water bath. The column outlet sample was collected from blood flowing out of the column outlet between 3.5 and 6.5 minutes after the start of the flow, with the time at which blood flowed into the column being set as 0 minutes.

[0119] For the obtained samples, the fluorescent labeled antibodies shown in Table 1 were used to stain the surface antigens of the platelets. Further, after adding 0.1% by mass PFA solution to fix the cells, the cells were kept in an ice-cold, dark place and the number of cells contained in each sample was rapidly determined. It should be noted that the platelet count was determined using a multi-item automatic blood cell analyzer XT-1800i (Sysmex company system). Surface antigen determination was performed using flow cytometry (BD Cytometer Setup and Tracking Beads, Becton, Dickinson and Company system).

[0120] The analysis was performed using BDFACS Diva (registered trademark) software Version 6.1.3 (manufactured by Becton, Dickinson and Company) or FLOWJO (manufactured by Tommy Digital Biology Co., Ltd.). The ratio of activated platelets was calculated by the following formula (5), the ratio of non-activated platelets was calculated by the following formula (6), the concentration of activated platelets was calculated by the following formula (7), and the concentration of non-activated platelets was calculated by the following formula (8). Using the obtained values, the activated platelet removal rate, non-activated platelet removal rate, and activated platelet removal ratio were calculated by the following formulas (9), (10), and (11), respectively.

[0121] Ratio of activated platelets (%) = Number of CD62-positive cells in the platelet fraction among CD41-positive cells / Number of platelets among CD41-positive cells × 100 Formula (5)

[0122] Ratio of non-activated platelets (%) = Number of CD62-negative cells in the platelet fraction among CD41-positive cells / Number of platelets among CD41-positive cells × 100 Formula (6)

[0123] Activated platelet concentration (cells / μL) = platelet number × ratio of activated platelets / 100 ···Formula (7)

[0124] Non-activated platelet concentration (cells / μL) = platelet number × ratio of non-activated platelets / 100 ···Formula (8)

[0125] Activated platelet removal rate (%) = {(activated platelet concentration at the column inlet side) - (activated platelet concentration at the column outlet side)} / (activated platelet concentration at the column inlet side) × 100 ···Formula (9)

[0126] Non-activated platelet removal rate (%) = {(non-activated platelet concentration at the column inlet) - (non-activated platelet concentration at the column outlet)} / (non-activated platelet concentration at the column inlet) × 100 ···Formula (10)

[0127] Activated platelet removal ratio = activated platelet removal rate / non-activated platelet removal rate ···Formula (11)

[0128] The activated platelet removal rate and the non-activated platelet removal rate were rounded off to the first decimal place, and the activated platelet removal ratio was rounded off to the second decimal place.

[0129] Here, as Figure 2As shown, the platelet fraction in CD41-positive cells means a platelet fraction 3 obtained by removing a microparticle fraction 4 in which platelets are finely fragmented and a platelet fraction 2 bound to red blood cells, etc., based on the cell population when the CD41-positive cell population is expanded using forward scattered light (FSC) on the horizontal axis and side scattered light (SSC) on the vertical axis.

[0130] [Table 1]

[0131] [Table 1]

[0132]

[0133] (Determination of PDGF Removal Rate)

[0134] The thickness of the water-insoluble carrier was measured using a micrometer (CLM2-10QMB or CLM2-10QMX, manufactured by Mitutoyo Co., Ltd.), and the volume of the knitted fabric was calculated using the following formula (12).

[0135] Next, four pieces of knitted fabric punched out to a diameter of 8 mm were placed in a polypropylene container. Inactivated fetal bovine serum (hereinafter referred to as "FBS") prepared to a concentration of 1000 pg / mL of PDGF (Product No. 220-BB, manufactured by R&D Systems, Inc.) was added to the container (inactivation conditions: 56°C, immersion for 2 hours) to adjust the volume of the knitted fabric relative to the liquid to 0.12 cm. 3 / mL, rotated at 27 rpm in a rotator in a 37°C incubator, and mixed by inversion for 24 hours. In addition, a blank was prepared by mixing FBS prepared in the same manner in a 37°C incubator for 24 hours by inversion except that no knitted fabric was added to the container.

[0136] The PDGF concentration in the blank and the inverted mixed FBS was measured using an ELISA kit (Product No. DBB00, manufactured by R&D Systems, Inc.), and the PDGF removal rate was calculated according to the following formula (13).

[0137] Volume of knitted fabric (cm 3 ) = (diameter of the knitted fabric to be punched out / 2) 2 ×π×thickness of knitted fabric×number of sheets···Formula (12)

[0138] PDGF removal rate (%) = {(PDGF concentration in the blank) - (PDGF concentration after inversion mixing)} / (PDGF concentration in the blank) × 100 ··· Formula (13)

[0139] In addition, the volume of the knitted fabric was rounded off to the third decimal place, and the PDGF removal rate was rounded off to the first decimal place.

[0140] (Determination of VEGF Removal Rate)

[0141] The thickness of the water-insoluble carrier was measured using a micrometer (CLM2-10QMB or CLM2-10QMX, manufactured by Mitutoyo Co., Ltd.), and the volume of the knitted fabric was calculated using Formula (12).

[0142] Next, four pieces of knitted fabric punched out to a diameter of 8 mm were placed in a polypropylene container. Inactivated fetal bovine serum (hereinafter referred to as "FBS") prepared to a concentration of 1000 pg / mL of VEGF (Product No. 293-VE / CF, manufactured by R&D Systems, Inc.) was added to the container so that the volume of the knitted fabric relative to the liquid volume was 0.12 cm. 3 / mL, rotated at 27 rpm in a rotator in a 37°C incubator, and mixed by inversion for 24 hours. In addition, a blank was prepared by mixing FBS prepared in the same manner in a 37°C incubator for 24 hours by inversion except that no knitted fabric was added to the container.

[0143] The VEGF concentration in the blank and the inverted mixed FBS was measured using an ELISA kit (Product No.: DVE00, manufactured by R&D Systems, Inc.), and the VEGF removal rate was calculated according to the following formula (14).

[0144] VEGF removal rate (%) = {(VEGF concentration in the blank) - (VEGF concentration after inversion mixing)} / (VEGF concentration in the blank) × 100 ··· Formula (14)

[0145] In addition, the volume of the knitted fabric was rounded off to the third decimal place, and the VEGF removal rate was rounded off to the first decimal place.

[0146] (Determination of FGF Removal Rate)

[0147] The thickness of the water-insoluble carrier was measured using a micrometer (CLM2-10QMB or CLM2-10QMX, manufactured by Mitutoyo Co., Ltd.), and the volume of the knitted fabric was calculated using the following formula (12). Next, four pieces of knitted fabric punched out to a diameter of 8 mm were placed in a polypropylene container. Normal human serum (Product No.: 12181201, Cosmo Bio Co., Ltd.) prepared with a concentration of 10,000 pg / mL of FGF (Product No.: 230-00791, manufactured by Ray Biotech) was added to the container so that the volume of the knitted fabric relative to the amount of liquid was 0.12 cm 3 / mL, rotated at 27 rpm in a rotator in a 37°C incubator, and mixed by inversion for 1 hour. In addition, normal human serum prepared in the same manner was mixed by inversion for 1 hour in a 37°C incubator, except that no knitted fabric was added to the container.

[0148] The FGF concentration in the blank and reverse mixed normal human serum was measured using an ELISA kit (Product No.: ELH-bFGF, manufactured by RayBiotech), and the FGF removal rate was calculated according to the following formula (15).

[0149] FGF removal rate (%) = {(FGF concentration in the blank) - (FGF concentration after inversion mixing)} / (FGF concentration in the blank) × 100 Formula (15)

[0150] In addition, the volume of the knitted fabric was rounded off to the third decimal place, and the FGF removal rate was rounded off to the first decimal place.

[0151] (Production of knitted fabrics)

[0152] Using the following components, sea-island composite fibers (fiber diameter: 3 dtex, 20 μm) having 704 islands per filament were bundled into 36 filaments under spinning conditions of a spinning speed of 1250 m / min to obtain fibers.

[0153] Island composition: Polypropylene

[0154] Sea ingredients: Polystyrene

[0155] Composite ratio (mass ratio): island: sea = 50:50

[0156] The obtained fiber was flat-knitted using a knitting machine (model name: circular knitting machine MR-1, Maruzen Industrial Co., Ltd.) to produce a knitted fabric with a unit area weight of 70 g / m 2 knitted fabric.

[0157] (Example 1)

[0158] (Production of Chloroacetamide Methylated Knitted Fabrics)

[0159] 3.3 g of NMCA was added to a mixture of 26 mL of nitrobenzene and 17 mL of 98% by mass sulfuric acid, and stirred at 10°C until the NMCA dissolved, preparing an NMCA solution. Next, 0.2 g of PFA was added to a mixture of 2 mL of nitrobenzene and 1.3 mL of 98% by mass sulfuric acid, and stirred at 20°C until the PFA dissolved, preparing a PFA solution. This 3.3 mL of PFA solution was cooled to 5°C and mixed with 43 mL of the NMCA solution to prepare Mixed Solution A. After stirring the mixture for 5 minutes, 1 g of the knitted fabric was added and impregnated for 0.25 hours. The impregnated knitted fabric was immersed in 43 mL of nitrobenzene at 10°C to stop the reaction. The nitrobenzene adhering to the fabric was then washed off with methanol and then rinsed by immersion in water to obtain Knitted Fabric A.

[0160] (Example 2)

[0161] A knitted fabric B was obtained by the same operation as in Example 1 except that the impregnation time of the knitted fabric in the mixed solution A was changed to 0.5 hours during the preparation of the chloroacetamide methylated knitted fabric.

[0162] (Example 3)

[0163] Knitted fabric C was obtained by carrying out the same operation as in Example 1 except that the impregnation time of the knitted fabric in the mixed solution A was changed to 1 hour during the preparation of the chloroacetamide methylated knitted fabric.

[0164] (Example 4)

[0165] Knitted fabric D was obtained by the same operation as in Example 1 except that the immersion time of the knitted fabric in the mixed solution A during the preparation of the chloroacetamide methylated knitted fabric was changed to 1.5 hours.

[0166] (Example 5)

[0167] Knitted fabric E was obtained by the same operation as in Example 1 except that the immersion time of the knitted fabric in the mixed solution A was changed to 2 hours during the preparation of the chloroacetamide methylated knitted fabric.

[0168] (Comparative Example 1)

[0169] Knitted fabric F was obtained by carrying out the same operation as in Example 1 except that the impregnation time of the knitted fabric in the mixed solution A was changed to 4 hours during the preparation of the chloroacetamide methylated knitted fabric.

[0170] (Example 6)

[0171] (Production of Tetraethylenepentamine-p-Chlorophenylated Knitted Fabric)

[0172] To a mixture of 1.5 mL of TEPA and 2.9 mL of triethylamine dissolved in 40 mL of dimethyl sulfoxide (hereinafter referred to as "DMSO"), the chloroacetamidomethylated knitted fabric A obtained in Example 1, which had been washed with methanol, was directly added and impregnated at 40°C for 3 hours. The knitted fabric was filtered out using a glass filter and washed with 40 mL of DMSO. Subsequently, 0.1 g of p-chlorophenyl isocyanate was added to 25 mL of DMSO, which had been dehydrated and dried over activated molecular sieves 3A, under a nitrogen atmosphere, and the mixture was heated to 30°C. The entire amount of the washed chloroacetamidomethylated knitted fabric was added and impregnated for 1 hour. The knitted fabric was filtered out using a glass filter, washed by immersion in the same amount of DMSO as the reaction solution, then washed by immersion in methanol, and then washed by immersion in water to obtain knitted fabric G.

[0173] (Example 7)

[0174] Knitted fabric H was obtained by the same operation as in Example 6 except that the knitted fabric used in the preparation of the tetraethylenepentamine-parachlorophenylated knitted fabric was made into knitted fabric B after washing with methanol.

[0175] (Example 8)

[0176] Knitted fabric I was obtained by carrying out the same operation as in Example 6 except that the knitted fabric in the preparation of the tetraethylenepentamine-p-chlorophenylated knitted fabric was made into knitted fabric C after washing with methanol.

[0177] (Example 9)

[0178] Knitted fabric J was obtained by carrying out the same operation as in Example 6 except that the knitted fabric in the preparation of the tetraethylenepentamine-p-chlorophenylated knitted fabric was made into knitted fabric D after washing with methanol.

[0179] (Example 10)

[0180] Knitted fabric K was obtained by the same operation as in Example 6 except that the knitted fabric used in the preparation of the tetraethylenepentamine-p-chlorophenylated knitted fabric was made into knitted fabric E after washing with methanol.

[0181] (Example 11)

[0182] Knitted fabric L was obtained by carrying out the same operation as in Example 7 except that the amount of TEPA added during the preparation of the tetraethylenepentamine-parachlorophenylated knitted fabric was changed to 0.5 mL.

[0183] (Example 12)

[0184] A knitted fabric M was obtained by carrying out the same operation as in Example 7 except that the amount of TEPA added during the preparation of the tetraethylenepentamine-parachlorophenylated knitted fabric was changed to 2 mL.

[0185] (Example 13)

[0186] Knitted fabric N was obtained by carrying out the same operation as in Example 7 except that the amount of TEPA added during the preparation of the tetraethylenepentamine-parachlorophenylated knitted fabric was changed to 4 mL.

[0187] (Example 14)

[0188] Knitted fabric O was obtained by carrying out the same operation as in Example 7 except that the amount of TEPA added during the preparation of the tetraethylenepentamine-parachlorophenylated knitted fabric was changed to 6 mL.

[0189] (Example 15)

[0190] (Production of Sulfonated Chloroacetamide Methylated Knitted Fabric)

[0191] 3.3 g of NMCA was added to a mixture of 26 mL of nitrobenzene and 17 mL of 98% by mass sulfuric acid, and stirred at 10°C until the NMCA dissolved, preparing an NMCA solution. Next, 0.2 g of PFA was added to a mixture of 2 mL of nitrobenzene and 1.3 mL of 98% by mass sulfuric acid, and stirred at 20°C until the PFA dissolved, preparing a PFA solution. 3.3 mL of this PFA solution was mixed with 43 mL of the NMCA solution and heated to 40°C to prepare a mixed solution A. After stirring the mixture for 5 minutes, 1 g of the knitted fabric was added and impregnated for 0.25 hours to allow the sulfonation reaction to proceed. The impregnated knitted fabric was immersed in 43 mL of nitrobenzene at 10°C to stop the reaction. The nitrobenzene adhering to the fabric was then washed off with methanol and then rinsed by immersion in water, obtaining a knitted fabric P.

[0192] (Example 16)

[0193] Knitted fabric Q was obtained by performing the same operation as in Example 7 except that the amount of TEPA added during the preparation of the tetraethylenepentamine-parachlorophenylated knitted fabric was changed to 10 mL.

[0194] (Example 17)

[0195] Knitted fabric R was obtained by carrying out the same operation as in Example 7 except that the amount of TEPA added during the preparation of the tetraethylenepentamine-parachlorophenylated knitted fabric was changed to 30 mL.

[0196] The resulting knitted fabrics A to R were measured for Rku, fiber diameter, activated platelet and non-activated platelet removal rates, PDGF removal rates, VEGF removal rates, and FGF removal rates using the above-described methods. Furthermore, the positive charge values ​​of knitted fabrics A to O and knitted fabrics Q to R were measured, while the negative charge value of knitted fabric P was measured, and the absolute values ​​were determined. The results are shown in Tables 2 and 3.

[0197] [Table 2]

[0198] [Table 2]

[0199]

[0200] [Table 3]

[0201] [Table 3]

[0202] VEGF removal rate FGF removal rate unit ％ ％ Example 1 - - Example 2 3 2 Example 3 - - Example 4 - - Example 5 - - Comparative Example 1 - - Example 6 64 50 Example 7 66 55 Example 8 73 67 Example 9 67 60 Example 10 62 53 Example 11 10 10 Example 12 81 68 Example 13 65 54 Example 14 38 33 Example 15 56 43 Example 16 25 23 Example 17 11 10

[0203] The above experimental results show that the water-insoluble carrier of the present invention has excellent activated platelet removal performance. Furthermore, it is shown that growth factors can also be adsorbed on water-insoluble carriers with a charge on their surface.

[0204] Industrial Applicability

[0205] The water-insoluble carrier of the present invention can remove activated platelets and can be used in the treatment of diseases for which therapeutic effects can be expected by inhibiting thrombosis.

[0206] Explanation of symbols

[0207] 1 Analysis range of one field of view in kurtosis measurement

[0208] 2. Platelet fraction bound to erythrocytes and other cells in CD41-positive cells

[0209] 3 Platelet fraction in CD41-positive cells

[0210] 4 Microparticle fraction of CD41-positive cells

[0211] l Reference length

Claims

1. A water-insoluble carrier having a surface kurtosis of 0.1 to 16.0 and capable of removing activated platelets.

2. The water-insoluble carrier according to claim 1, wherein The surface carries an electrical charge.

3. The water-insoluble carrier according to claim 1 or 2, wherein The surface charge is 0.3 to 3.0 mmol relative to 1 g of dry mass.

4. The water-insoluble carrier according to any one of claims 1 to 3, wherein The surface has amino groups. The water-insoluble carrier according to any one of claims 1 to 4, which is in the form of fibers or particles. The water-insoluble carrier according to claim 5 , which has a diameter of 1 to 100 μm. 7 . The water-insoluble carrier according to claim 1 , comprising a polymer selected from the group consisting of polystyrene, polypropylene, polysulfone, polyethersulfone and cellulose. 8 . The water-insoluble carrier according to claim 1 , wherein platelet-derived growth factor, vascular endothelial growth factor and / or fibroblast growth factor are removed.

9. A blood purifier comprising the water-insoluble carrier according to any one of claims 1 to 8.

10. The blood purifier according to claim 9, which is a blood purification column. The blood purifier according to claim 10 , which is used for inhibiting thrombosis.

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