JP7290132B2 - Material for removal of activated granulocyte-activated platelet complexes - Google Patents

Description

The present invention relates to materials for removing activated granulocyte-activated platelet complexes.

Until now, cytokines have been used as therapeutic methods for inflammatory diseases such as acute respiratory distress syndrome (ARDS), acute lung injury (ALI), systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, ulcerative colitis, and Crohn's disease. Extracorporeal circulation therapy using materials capable of removing blood components such as leukocytes and platelets, which are sources of inflammatory substances, from the body is used.

In the case of extracorporeal circulation therapy, removal of lymphocytes from the blood of the treated patient generally lowers the immune function of the treated patient, leading to the risk of developing an infectious disease during or after the treatment. Therefore, when a material for removing blood components is used for extracorporeal circulation therapy, it is desirable that the amount of lymphocytes removed by the material is small.

In recent years, activated granulocyte-activated platelet complexes have attracted attention as a new causative substance that causes inflammatory diseases. The activated granulocyte-activated platelet complex has a higher ability to migrate to tissues exhibiting inflammatory reactions compared to granulocytes alone or activated granulocytes alone, and the interaction between activated granulocytes and activated platelets is high. Its action has been reported to increase the release of inflammatory substances compared to granulocytes alone or activated granulocytes alone (Non-Patent Document 1).

As a material that can be used for extracorporeal circulation therapy, for example, Patent Document 1 discloses a material for removing activated granulocyte-activated platelet complexes with a defined amount of amide groups.

Patent Document 2 discloses a blood component adsorption column in which the open area ratio and packing density of the knitted fabric are specified.

Patent Document 3 discloses a material for blood purification that has the ability to remove activated granulocyte-activated platelet complexes with defined amounts of amide groups and amino groups.

US Pat. No. 5,300,001 discloses porous particles made of a composition containing an ethylene-vinyl alcohol copolymer for producing a material for selective removal of granulocytes and monocytes.

JP 2019-136499 A JP 2018-102659 A WO2018/047929 JP 2010-233825 A

Alexander Zarbock et al., J. Clin. Invest. 2006, vol. 116, p. 3211-3219

However, the average open area of the fibers used in the examples of Patent Document 1 is estimated to be about 0.041 to 0.045 mm ² from the manufacturing method, and lymphocytes are removed when used for extracorporeal circulation therapy. it is conceivable that. Further, in Patent Document 1, blood cell removal is controlled by a specific amide group structure, but there is no description about the average pore area, and there is no relationship between the amount of amide groups and the average pore area, and the average pore area. No association with leukocyte removal performance is suggested. Furthermore, Patent Document 1 does not contain any description related to suppression of removal of lymphocytes.

The blood component adsorption column described in Patent Document 2 describes lymphocytes (white blood cells) as objects to be removed, and there is no concept of suppressing the removal of lymphocytes. Furthermore, the average open area of the fibers used in the examples is estimated to be 0.079 mm ² or less from the manufacturing method. Patent Document 2 does not describe the relationship between the blood component adsorption column and the activated granulocyte-activated platelet complex, nor does it describe the removal of the activated granulocyte-activated platelet complex. In addition, leukocytes and activated granulocyte-activated platelet complexes differ in the types of proteins expressed on the surface of blood cells, and the size of the blood cells also differs. It is thought that the mechanism is not the same when removing the complex.

The material for blood purification described in Patent Document 3 describes leukocyte components as objects to be removed, and has no idea of suppressing the removal of lymphocytes. Furthermore, the average open area of the fibers used in the examples is estimated to be about 0.008 to 0.027 mm ² from the manufacturing method, and it is believed that lymphocytes are removed when the fibers are used for extracorporeal circulation therapy.

The removal material described in Patent Document 4 is characterized by being porous particles, and there is no concept of average open pore area. In addition, granulocytes and monocytes and activated granulocyte-activated platelet complexes differ in the types of proteins expressed on the surface of blood cells and also in the size of the blood cells. It is thought that the mechanism is not the same in removing activated granulocyte-activated platelet complexes.

Under the above background, development of a material capable of suppressing removal of lymphocytes and removing activated granulocyte-activated platelet complexes is desired.

Accordingly, an object of the present invention is to provide a material capable of suppressing removal of lymphocytes and selectively removing activated granulocyte-activated platelet complexes.

In order to solve the above problems, the present inventors conducted studies and found that a fibrous material having a ligand containing an amide group and having a specific average pore area formed activated granulocyte-activated platelet complexes. We have found that it can be removed selectively.

That is, the present invention includes the following [1] to [6].
[1] A knitted fabric or woven fabric made of fibers having a ligand containing an amide group is included, and the average open area per stitch of the knitted fabric or the texture of the fabric is 0.080 mm ² or more and 0.100 mm ² Removed materials for activated granulocyte-activated platelet complexes, which are:
[2] The removal material according to [1], wherein the knitted fabric is a weft knitted fabric, and the aspect ratio of the openings in the stitches is 0.75 to 1.30.
[3] The removal material according to [1] or [2], wherein the fiber contains polystyrene or a polystyrene derivative.
[4] The fiber is a sea-island composite fiber, the sea component of the sea-island composite fiber is polystyrene or a polystyrene derivative, and the island component of the sea-island composite fiber is polypropylene. A removal material according to any one of the preceding claims.
[5] The removal material according to any one of [1] to [4], wherein the average open area is 0.083 mm ² or more and 0.089 mm ² or less.
[6] A blood purifier comprising the removal material according to any one of [1] to [5].

The removal material of the present invention can suppress the removal of lymphocytes contained in the blood of patients with inflammatory diseases, etc., and selectively remove activated granulocyte-activated platelet complexes.

FIG. 10 shows the apertures in the knitted fabric-shaped activated granulocyte-activated platelet complex removal material. FIG. 10 shows the apertures in the fabric-shaped activated granulocyte-activated platelet complex removal material. FIG. 10 is a diagram showing a method of calculating the aspect ratio of the apertures in the knitted fabric-shaped material from which the activated granulocyte-activated platelet complexes have been removed.

The present invention will be described in detail below.

The material for removing activated granulocyte-activated platelet complexes of the present invention comprises a knitted fabric or fabric comprising fibers having a ligand containing an amide group, and The average open area is characterized by being 0.080 mm ² or more and 0.100 mm ² or less.

A "knitted fabric" is a structure made by knitting fibers. There is no particular limitation on the knitting method of the knitted fabric. Examples include weft knitted fabrics (flat knitted fabric, rubber knitted fabric, pearl knitted fabric) and warp knitted fabrics (single tricot knitted fabric, single cord knitted fabric, single atlas knitted fabric). However, the weft knitted fabric is preferable from the viewpoint of the shape of the openings suitable for permeation of blood cells.

A "fabric" is a structure made by weaving fibers. There is no particular limitation on the weaving method of the fabric. For example, a plain weave fabric, a twill weave fabric, a dutch weave fabric, and a twilled dutch weave fabric may be mentioned, but from the viewpoint of the shape of the openings suitable for permeation of blood cells, a plain weave fabric or a twill weave fabric is preferable.

A "stitch" is a structure surrounded by a combination of loops of fibers that constitute a knitted fabric, and has an opening.

A "texture" is a structure formed by being surrounded by warp and weft threads that constitute a fabric, and has openings.

"Aperture part" refers to a region where there are no fibers constituting a knitted or woven fabric when viewed from directly above a piece of knitted or woven fabric grounded on a plane, and one stitch or The weave has one aperture. For example, "1" and "2" in FIG. 1 and "4" in FIG. 2 are apertures. Here, a single knitted or woven fabric grounded on a flat surface refers to a knitted or woven fabric that does not warp, overlap, twist or break even if any part of the fibers that make up the knitted or woven fabric. . In addition, a portion including an end portion of a knitted or woven fabric is not considered to be an "opening portion" even if the portion does not contain fibers.

The “average open area” is the average area of open holes per stitch of a knitted fabric or per stitch of a woven fabric. The number of apertures measured when calculating the average aperture area should be 100 or more. A method for measuring the average open area is shown below. An image of a single knitted fabric or woven fabric grounded on a flat surface is obtained from directly above, and the image analysis software ImageJ (National Institutes of Health, USA) is used from the acquired image. Analyze the area of the perforation per eye. A method of analyzing the area of the aperture is as follows. First, by binarizing the image with the "Threshold" function, only the apertures are extracted. Next, select the range to be analyzed and execute “Analyze Particles (Size is set to 0.001 mm ² to 0.5 mm ² )”. The average value (output as Average Size) of the calculated areas of the pore portions (output as Area) is taken as the average pore area. A range that includes the ends of knitted fabrics or woven fabrics should not be selected as the range to be analyzed. Here, the high ratio of removal rate of activated granulocyte-activated platelet complexes to removal rate of lymphocytes makes it possible to achieve extracorporeal circulation treatment with reduced risk. Therefore, the average open area should be 0.080 mm ² or more and 0.100 mm ² or less, preferably 0.083 mm ^{2 or} more and 0.089 mm ² or less.

The term "opening ratio" refers to the ratio of openings in the field of view when a single knitted or woven fabric that is grounded on a flat surface is viewed from directly above. In addition, 100 or more apertures must be included in the field of view when the aperture ratio is measured. A method for measuring the porosity is shown below. An image of a piece of knitted fabric or woven fabric grounded on a flat surface is obtained as viewed from directly above, and the porosity is calculated from the obtained image using image analysis software ImageJ (National Institutes of Health, USA). First, by binarizing the image with the "Threshold" function, only the apertures are extracted. Next, the range to be analyzed is selected, and "Analyze Particles (Size is set to 0.001 mm ² to 0.5 mm ² )" is executed to obtain the pore area. The ratio of the total open area (output as Total Area) to the area of the analysis range is defined as the open area ratio (output as %Area). A range that includes the ends of knitted fabrics or woven fabrics should not be selected as the range to be analyzed.

The "aspect ratio of the openings" is the average value obtained by dividing the length of the openings in the wale direction by the length of the openings in the course direction. Here, the length of the opening in the wale direction is the maximum value of the length of a straight line parallel to the wale direction that can be drawn in the opening (“8” in the case of the opening “7” in FIG. 3). length). The length of the opening in the course direction is the maximum value of the length of a straight line parallel to the course direction that can be drawn in the opening (the length of "9" in the case of opening "7" in FIG. 3). ). Therefore, the aspect ratio of the openings can also be said to be the average value of the values obtained by dividing the maximum length of the openings in the wale direction by the maximum length of the openings in the course direction. For ten randomly selected apertures, the length in the wale direction of each aperture and the length in the course direction of the aperture were measured, and the length in the wale direction of the aperture was measured. A value obtained by dividing the length of the hole in the course direction is calculated, and the average value of the obtained 10 values is taken as the aspect ratio of the hole. The aspect ratio of the apertures is preferably 0.75 to 1.30, more preferably 0.85 to 1.15, from the viewpoint of removal performance.

The average pore area, pore opening ratio, and aspect ratio of the pore portions of the knitted fabric can be controlled by the needle density of the knitting needles of the knitting machine used for knitting and the value of the stitch adjustment scale. For example, when the needle density is lowered, the length of the openings of the knitted fabric in the course direction is shortened, so that the average opening area and the opening ratio are decreased, and the aspect ratio of the openings is increased. In addition, when the value of the density adjustment scale is increased, the length of the openings of the knitted fabric in the wale direction increases, so the average opening area and opening ratio increase, and the aspect ratio of the openings also increases.

The average open area and open area of the fabric can be controlled by warp density, weft density and fiber diameter. For example, as a method of increasing the average open area or opening ratio, there is a method of decreasing the warp density, decreasing the weft density, and decreasing the fiber diameter.

The fibers that make up the knitted or woven fabric may be single component fibers or multicomponent fibers. The fiber preferably contains polystyrene or a polystyrene derivative as a component thereof, because of the ease of imparting ligands to the fiber. Here, the polystyrene derivative refers to a polymer in which a part of polystyrene (eg, benzene ring) is substituted with a functional group. -methylstyrene, poly-p-tert-butoxystyrene, poly-p-acetoxystyrene, poly-p-(1-ethoxyethoxy)styrene and the like. When polystyrene or a polystyrene derivative is included as a fiber component, it is preferable to use a reinforcing material from the viewpoint of workability.

The form of the fiber using the reinforcing material is not particularly limited, and may be a fiber obtained by blending the reinforcing material with other components, a core-sheath fiber using the reinforcing material as a core material, or a sea-island composite fiber using the reinforcing material as an island component. However, from the viewpoint of securing the adsorption of the removed substances to the fiber surface, core-sheath fibers with a reinforcing material as a core material and sea-island composite fibers with a reinforcing material as an island component are preferable, and the effect of improving the strength of the fiber is High sea-island composite fibers are more preferred.

A sea-island composite fiber is a fiber having a cross-sectional structure in which island components made of a certain polymer are interspersed in sea components made of a different polymer.

The sea component means a polymer existing on the surface side of the sea-island composite fiber.

When the fiber is a sea-island composite fiber, the sea component is preferably polystyrene or a polystyrene derivative.

The island component means a polymer different from the sea component scattered in the sea component when viewed in the direction perpendicular to the fiber axis direction of the sea-island composite fiber.

When the fiber is a sea-island composite fiber, the island component is preferably selected from polypropylene, polycarbonate, or polyethylene from the viewpoint of ensuring the strength of the fiber. A preferred embodiment of the sea-island composite fiber includes, for example, a sea-island composite fiber in which the sea component is polystyrene or a polystyrene derivative and the island component is polypropylene.

By "ligand containing an amide group" is meant a chemical structure containing an amide group attached to fibers to confer ability to clear activated granulocyte-activated platelet complexes. The structure of the ligand containing an amide group is not particularly limited as long as it is a chemical structure containing at least one amide group. Therefore, it is desirable that the chemical structure adjacent to the amide group does not contain a cyclic structure (eg, aromatic hydrocarbon or cyclic hydrocarbon and its substitutes). As a preferred embodiment of the ligand containing an amide group, the chemical structure adjacent to the amide group is an alkyl group optionally substituted with a halogen atom, that is, the ligand containing an amide group is an acetamidoalkyl group (e.g., acetamidomethyl group) and halogenated acetamidoalkyl groups (eg, chloroacetamidomethyl group, chloropropionamidomethyl group, chloroacetamidoethyl group, etc.), and chloroacetamidomethyl group is preferred.

As a method for imparting a ligand containing an amide group to a fiber, there is a method of fiberizing a polymer having a ligand containing an amide group, and a method of fiberizing the polymer after the polymerization reaction and then introducing the ligand into the obtained fiber. However, a method of introducing a ligand into the resulting fiber after fiberizing the polymer after the polymerization reaction is preferred. When fiberizing a polymer having a ligand containing an amide group, a monomer containing an amide group in the side chain structure may be polymerized and the resulting polymer may be fiberized, or the amide group may be bound after the polymerization reaction, followed by It may be fibrous. As a method of fiberizing a polymer having a ligand containing an amide group, for example, polystyrene and N-hydroxymethyl-2-chloroacetamide are reacted to synthesize polystyrene to which a chloroacetamidomethyl group is bound, and then the synthesized chloro A method of fiberizing acetamidomethylated polystyrene is included. When binding a ligand to the fiber obtained after making the polymer after the polymerization reaction into a fiber, there are a method of binding a ligand containing an amide group to the fiber and a method of generating an amide bond by reacting the fiber with a compound. However, the method of attaching a ligand containing an amide group to the fiber is preferred. As a method for binding a ligand containing an amide group to a fiber, for example, there is a method of reacting a halogenated hydrocarbon derivative containing an amide group with the surface of a fiber having an aromatic hydrocarbon structure. For example, in the case of a sea-island composite fiber in which the sea component is polystyrene and the island component is polypropylene, by reacting polystyrene with N-hydroxymethyl-2-chloroacetamide, the chloroacetamidomethyl group, which is a ligand, is converted to polystyrene as the sea component. A bonded sea-island composite fiber can be obtained. As a method of generating an amide bond by reacting a fiber with a compound, there are a method of reacting an amine with a fiber surface having a carboxylic acid structure, a method of reacting a carboxylic acid with a fiber surface having an amine structure, and a method of reacting a nitrile fiber with an acid. Examples include a method of decomposition. In addition, further chemical modification can be carried out as long as it does not affect the ligand containing the amide group. For example, a polyamine such as tetraethylenepentamine can be added to the chloro group of polystyrene to which a chloroacetamidomethyl group has been added.

There is no particular limitation on the binding mode between the fiber and the ligand containing an amide group, but from the viewpoint of the stability of the binding with the fiber, the fiber and the ligand are bound by ionic bond, hydrogen bond or van der Waals bond. It is preferable that they are directly bonded by a covalent bond or an ionic bond. Ligands containing amide groups are preferably attached to the fiber surface.

In the present specification, the activated granulocyte-activated platelet complex is not particularly limited as long as it is a complex in which activated granulocytes and activated platelets are bound. In patients with inflammatory diseases, activated granulocyte-activated platelet complexes are thought to be directly involved in pathology through phagocytosis into self-tissues and release of cytokines. Therefore, removal of activated granulocyte-activated platelet complexes is considered necessary for the treatment of inflammatory diseases. For measuring the concentration of activated granulocyte-activated platelet complexes, for example, a fluid containing activated granulocyte-activated platelet complexes is added with an activation detection reagent that specifically binds to activated platelets, and an activated This is done by reacting an activated detection reagent that specifically binds to granulocytes and measuring the fraction of blood cells bound to both reagents. A flow cytometer can be used for the measurement.

“Activated granulocytes” means granulocytes that release cytokines, active oxygen, proteolytic enzymes, etc. by cytokines, lipopolysaccharide (hereinafter, LPS), etc. Examples thereof include activated neutrophils. The degree of activation can be determined by measuring the amount of cytokines, active oxygen, and proteolytic enzymes released by granulocytes. In addition, since the surface antigens of activated granulocytes and non-activated granulocytes are different, the expression of surface antigens peculiar to activated granulocytes can be determined by immunofluorescent staining or flow cytometry. , CD11b (activated)-positive granulocytes can be defined as activated granulocytes.

By "activated platelets" is meant platelets that release cytokines, adenosine diphosphate, serotonin and the like. The degree of activation can be determined by measuring the amount of cytokines released by platelets. In addition, since the surface antigens of activated platelets and non-activated platelets are different, it can be determined by measuring the expression of surface antigens specific to activated platelets by immunofluorescence staining, flow cytometry, etc. For example, CD62P positive Platelets can be activated platelets.

The material for removing activated granulocyte-activated platelet complexes of the present invention can be used as a carrier for an extracorporeal circulation column for removing activated granulocyte-activated platelet complexes.

Further, the present invention is characterized by providing a blood purifier comprising the activated granulocyte-activated platelet complex-removing material.

"Blood purifier" refers to a product that at least partially contains medical materials for the purpose of circulating blood outside the body and removing waste products and harmful substances from the blood. Extracorporeal circulation columns packed with fabric are included.

Furthermore, the blood purifier comprising the activated granulocyte-activated platelet complex-removing material can be suitably used for treatment of inflammatory diseases. When used for the treatment of inflammatory diseases, a patient is connected to the blood purifier provided with the material for removing activated granulocyte-activated platelet complexes by a blood circuit, and the blood purifier of the present invention is fed from the patient to the blood purifier. A method of extracorporeal blood circulation, in which the blood withdrawn is passed through and returned to the patient, is preferred.

The blood purifier comprising the activated granulocyte-activated platelet complex-removing material may be used in combination with other body fluid treatment methods, medical devices, and pharmaceuticals. Other body fluid treatment methods, medical devices, and pharmaceuticals include, for example, plasmapheresis, peritoneal dialysis, plasma separators, hemofilters, heart-lung machines or ECMO (extracorporeal membrane oxygenation), anticoagulants, platelet preparations, and vasopressors. .

As a method for evaluating the activated granulocyte-activated platelet complex-removing performance and the lymphocyte-removing performance of the material for removing activated granulocyte-activated platelet complexes, for example, activated granulocyte-activated A liquid containing activated platelet complexes and lymphocytes is impregnated with the removal material, and after impregnation, the amount of decrease in activated granulocyte-activated platelet complexes and lymphocytes in the liquid is evaluated, and their removal rates are calculated. method. In addition, a container having an inlet and an outlet is filled with the above-mentioned removal material, and a liquid containing activated granulocyte-activated platelet complexes and lymphocytes is passed through, and the change in their concentrations at the inlet and outlet causes them to be removed. can also be used.

The concentration of lymphocytes is measured, for example, by allowing a lymphocyte detection reagent to react with a liquid containing lymphocytes and measuring the blood cell fraction bound to the reagent. A flow cytometer or a hemocytometer can be used for the measurement.

The platelet concentration is measured, for example, by allowing a platelet detection reagent to react with a liquid containing platelets and measuring the blood cell fraction bound to the reagent. A flow cytometer or a hemocytometer can be used for the measurement.

The activated platelet detection reagent does not bind to non-activated platelets and granulocytes, but has binding properties to activated platelets. For example, anti-CD62P antibody that recognizes an activated platelet-specific cell surface marker). can be used. In addition, the activated granulocyte detection reagent does not bind to non-activated granulocytes and platelets, but has binding properties to activated granulocytes, and contains cell surface markers specific to or common to desired leukocyte components. Antibodies can be mentioned, and an anti-CD11b antibody can be used as a detection reagent for activated granulocytes, for example. Among others, activated granulocytes can be specifically detected by using an activated anti-CD11b antibody that can specifically detect the activated conformation. Also, an anti-CD45 antibody can be used to detect granulocytes and lymphocytes. For the detection of granulocytes, forward scattered light and side scattered light may be combined, anti-CD45 antibody and side scattered light may be combined, or anti-CD45 antibody and anti-CD66b antibody may be combined. For detection of lymphocytes, forward scattered light and side scattered light may be combined, anti-CD45 antibody and side scattered light may be combined, anti-CD4 antibody or anti-CD8 antibody may be combined, and A cell population obtained by subtracting CD66b-positive cells and CD14-positive cells from CD45-positive cells can also be defined as lymphocytes. An anti-CD41 antibody can be used to detect platelets.

Preferably, the detection reagent is labeled with an indicator for confirmation of binding. The index is arbitrarily selected according to the detection method employed. Measurement by a flow cytometer is preferably used from the viewpoint of ease of operation and quantification, but in this case the detection reagent is fluorescently labeled. The fluorescent label is not particularly limited, and for example, FITC (fluorescein isothiocyanate), PE (phycoerythrin), or APC (allophycocyanin) labeling can be employed. The activated granulocyte detection reagent and the activated platelet detection reagent are labeled with different fluorescent substances. These labeled detection reagents can be produced according to conventional methods, and are also available as commercial products.

The material for removing activated granulocyte-activated platelet complexes of the present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.

(spinning)
A fiber (hereinafter referred to as fiber A) was obtained by bundling 36 filaments of sea-island fibers (fineness: 3 dtex, fiber diameter: 20 μm) with 256 islands per filament at a spinning speed of 1250 m/min using the following components. .
Island component: Polypropylene Sea component: Polystyrene Composite ratio (weight ratio): Island: Sea = 50:50

(Production of knitted fabric A)
Using fiber A, adjust the thread adjustment scale of a tubular knitting machine (model name: circular knitting machine MR-1, Maruzen Sangyo Co., Ltd.) to 2.50, set the diameter of the hook of the tubular knitting machine to 7.00 inches, and the number of needles. was knitted with 480 needles (needle density: 8.6 needles/cm) to obtain a knitted fabric A.

(Production of knitted fabric B)
A knitted fabric B was obtained by performing the same operation as for the knitted fabric A, except that the stitch adjustment scale of the tubular knitting machine was adjusted to 2.75.

(Production of knitted fabric C)
A knitted fabric C was obtained by performing the same operation as for the knitted fabric A, except that the stitch adjustment scale of the tubular knitting machine was adjusted to 3.00.

(Production of knitted fabric D)
A knitted fabric D was obtained by performing the same operation as for the knitted fabric A, except that the stitch adjustment scale of the tubular knitting machine was adjusted to 3.50.

(Production of knitted fabric E)
A knitted fabric E was obtained by performing the same operation as for the knitted fabric A, except that the stitch adjustment scale of the tubular knitting machine was adjusted to 5.00.

(Production of knitted fabric F)
Knitted fabric F was obtained by performing the same operation as knitted fabric A except that the diameter of the hook of the tubular knitting machine was 5.25 inches and the number of needles was 252 (needle density: 6.0 / cm). Obtained.

(Production of knitted fabric G)
A knitted fabric G was obtained by performing the same operation as for the knitted fabric F, except that the stitch adjustment scale of the tubular knitting machine was adjusted to 2.75.

(Production of knitted fabric H)
A knitted fabric H was obtained by performing the same operation as for the knitted fabric F, except that the stitch adjustment scale of the tubular knitting machine was adjusted to 3.00.

(Production of knitted fabric I)
A knitted fabric I was obtained by performing the same operation as for the knitted fabric F, except that the stitch adjustment scale of the tubular knitting machine was adjusted to 3.50.

(Production of removal material A)
Nitrobenzene 46 wt%, sulfuric acid 46 wt%, paraformaldehyde 1 wt% and N-hydroxymethyl-2-chloroacetamide (hereinafter referred to as NMCA) 7 wt% were mixed at 10°C or less, stirred, and dissolved in a reaction solution (hereinafter referred to as NMCA reaction solution ) was prepared. 1.0 g of the knitted fabric A was added to 40 mL of the NMCA reaction liquid cooled to 5°C, and the reaction liquid was allowed to react for 2 hours while the reaction liquid was kept at 5°C. After that, the knitted fabric A was taken out from the reaction liquid, and the knitted fabric A was immersed in 40 mL of nitrobenzene and washed. Subsequently, the knitted fabric A was taken out, immersed in methanol and washed to obtain an amidated knitted fabric A (hereinafter referred to as removed material A).

(Preparation of removal material B)
Amidated knitted fabric B (hereinafter referred to as removed material B) was obtained by performing the same operation as in the manufacturing method of removed material A, except that knitted fabric A was changed to knitted fabric B.

(Preparation of removal material C)
An amidated knitted fabric C (hereinafter referred to as "removed material C") was obtained by performing the same operation as in the manufacturing method of the removed material A, except that the knitted fabric A was changed to the knitted fabric C.

(Preparation of removed material D)
Amidated knitted fabric D (hereinafter referred to as removed material D) was obtained by performing the same operation as in the production method of removed material A except that knitted fabric A was changed to knitted fabric D.

(Preparation of removal material E)
An amidated knitted fabric E (hereinafter referred to as "removed material E") was obtained by performing the same operation as in the manufacturing method of the removed material A, except that the knitted fabric A was changed to the knitted fabric E.

(Preparation of removal material F)
Amidated knitted fabric F (hereinafter referred to as removed material F) was obtained by performing the same operation as in the production method of removed material A, except that knitted fabric A was changed to knitted fabric F.

(Preparation of removal material G)
An amidated knitted fabric G (hereinafter referred to as removed material G) was obtained by performing the same operation as in the manufacturing method of the removed material A, except that the knitted fabric A was changed to the knitted fabric G.

(Preparation of removal material H)
Amidated knitted fabric H (hereinafter referred to as removed material H) was obtained by performing the same operation as in the manufacturing method of removed material A except that knitted fabric A was changed to knitted fabric H.

(Preparation of removal material I)
Amidated knitted fabric I (hereinafter referred to as removed material I) was obtained by performing the same operation as in the manufacturing method of removed material A except that knitted fabric A was changed to knitted fabric I.

(Example 1)
In order to confirm the ability of the removed material F to remove activated granulocyte-activated platelet complexes and lymphocytes, the removed material F was layered and filled in a column, brought into contact with healthy human blood for a predetermined period of time, and the solution before and after the contact was measured. The concentrations of activated granulocyte-activated platelet complexes and lymphocytes in the medium were measured. The measurement method is shown below.

After cutting out the removed material F into a disk shape with a diameter of 1 cm, the thickness was measured. In a cylindrical column (inner diameter 1 cm x height 1.2 cm, inner volume 0.94 cm ³ , outer diameter 2 cm, made of polycarbonate) with upper and lower liquid inlets and outlets, the product of the measured thickness and the number of packed columns is 0.98 cm. The removal material F cut out in the form of a disc was laminated and filled. When the number of filling sheets did not become an integer, the number after the decimal point was rounded off. Healthy human blood to which LPS was added to 70 EU/mL was shaken at 37° C. for 30 minutes at 65 rpm to activate blood cells, and then the blood was passed through the column at a flow rate of 0.63 mL/min. Then, blood was collected at the inlet side and the outlet side of the column. The blood on the outlet side of the column was collected at 3.5 to 6.5 minutes after the time when the blood flowed into the column was defined as 0 minutes. The blood obtained after the end of the passage was measured with a hemocytometer (Sysmex, multi-item automatic blood cell analyzer XT-1800i) to calculate the concentrations of granulocytes and lymphocytes. In addition, blood samples were prepared by staining the surface antigens of blood cells present in the blood after passage with the fluorescence-labeled antibodies shown in Table 1, followed by hemolysis using VersaLyse. Prepared blood samples were stored on ice in the dark until measurement using a flow cytometer. After that, by measurement using a flow cytometer (BD Cytometer Setup and Tracking Beads (Becton, Dickinson and Company)), the number of activated granulocyte-activated platelet complexes with granulocytes as the parameter in each blood sample was determined. Percentages were measured. The concentration of activated granulocyte-activated platelet complexes was calculated according to Equation 1. In addition, ethidium monoazide bromide was used for determination of viable cells. FLOWJO (Tomy Digital Biology Co., Ltd.) was used for the analysis. The removal rate of activated granulocyte-activated platelet complexes and the removal rate of lymphocytes when liquid was passed through the column were calculated by the following equations 2 and 3, respectively. Then, the ratio of the removal rate of the activated granulocyte-activated platelet complex and the removal rate of the lymphocyte was calculated according to Equation 4. Table 2 shows the results.

Concentration of activated granulocyte-activated platelet complexes = (concentration of granulocytes) x (percentage of activated granulocyte-activated platelet complexes when granulocytes are used as parameters) Formula 1

Removal rate (%) of activated granulocyte-activated platelet complex = {(concentration of activated granulocyte-activated platelet complex at column inlet side)-(activated granulocyte-activated platelet at column outlet side Concentration of complexes)}/(concentration of activated granulocyte-activated platelet complexes on column inlet side)×100 Equation 2

Lymphocyte removal rate (%)={(concentration of lymphocytes on column inlet side)−(concentration of lymphocytes on column outlet side)}/(concentration of lymphocytes on column inlet side)×100 Equation 3

Ratio of activated granulocyte-activated platelet complex removal rate to lymphocyte removal rate=(activated granulocyte-activated platelet complex removal rate)/(lymphocyte removal rate) Formula 4

(Example 2)
The removal material G was subjected to the same operation as in Example 1, and the removal rate of activated granulocyte-activated platelet complexes, the removal rate of lymphocytes, the removal rate of activated granulocyte-activated platelet complexes and the lymphocyte The ratio of sphere removal rate is shown. Table 2 shows the results.

(Example 3)
The removal material H was subjected to the same operation as in Example 1, and the removal rate of activated granulocyte-activated platelet complexes, the removal rate of lymphocytes, the removal rate of activated granulocyte-activated platelet complexes and the lymphocyte The ratio of sphere removal rate is shown. Table 2 shows the results.

(Comparative example 1)
The removal material A was subjected to the same operation as in Example 1, and the removal rate of activated granulocyte-activated platelet complexes, the removal rate of lymphocytes, the removal rate of activated granulocyte-activated platelet complexes and the lymphocyte The ratio of sphere removal rate is shown. Table 2 shows the results.

(Comparative example 2)
The removal material B was subjected to the same operation as in Example 1, and the removal rate of activated granulocyte-activated platelet complexes, the removal rate of lymphocytes, the removal rate of activated granulocyte-activated platelet complexes and the lymphocyte The ratio of sphere removal rate is shown. Table 2 shows the results.

(Comparative Example 3)
For the removed material C, the same operation as in Example 1 was performed, and the removal rate of activated granulocyte-activated platelet complexes, the removal rate of lymphocytes, the removal rate of activated granulocyte-activated platelet complexes and the lymphocyte The ratio of sphere removal rate is shown. Table 2 shows the results.

(Comparative Example 4)
For the removed material D, the same operation as in Example 1 was performed, and the removal rate of activated granulocyte-activated platelet complexes, the removal rate of lymphocytes, the removal rate of activated granulocyte-activated platelet complexes and the lymphocyte The ratio of sphere removal rate is shown. Table 2 shows the results.

(Comparative Example 5)
The removal material E was subjected to the same operation as in Example 1, and the removal rate of activated granulocyte-activated platelet complexes, the removal rate of lymphocytes, the removal rate of activated granulocyte-activated platelet complexes and the lymphocyte The ratio of sphere removal rate is shown. Table 2 shows the results.

(Comparative Example 6)
The removal material I was subjected to the same operation as in Example 1, and the removal rate of the activated granulocyte-activated platelet complex, the removal rate of the lymphocyte, the removal rate of the activated granulocyte-activated platelet complex and the lymphocyte The ratio of sphere removal rate is shown. Table 2 shows the results.

(Example 4)
The average open area, open area ratio, and aspect ratio of the open area of the removed material F were measured. The measurement method is shown below.

One sheet of removed material F was placed on a flat surface, and an image of the knitted fabric was obtained with an image scanner (EPSON, GT-X980). Acquired images were analyzed using ImageJ (National Institutes of Health). The analysis method is as follows. First, the image was binarized with the "Threshold" function to extract only the apertures. Next, the range to be analyzed was selected and "Analyze Particles (Size set to 0.001 mm ² to 0.5 mm ² )" was executed. A range including the edge of the knitted fabric should not be selected as the range to be analyzed. The average value (output as Average Size) of the calculated areas of the apertures (output as Area) is defined as the average aperture area, and the sum of the aperture areas occupying the area of the analysis range (output as Total Area) ) was taken as the hole opening ratio (output as %Area). Furthermore, the length of the aperture in the wale direction and the length of the aperture in the course direction were measured using ImageJ. For ten randomly selected apertures, the length of the aperture in the wale direction and the length of the aperture in the course direction were measured. A value divided by the length in the direction was calculated, and the average value of the obtained 10 values was taken as the aspect ratio of the aperture. Table 3 shows the results.

(Example 5)
The removal material G was subjected to the same operation as in Example 4, and the average open area, open area ratio, and aspect ratio of open areas were measured. Table 3 shows the results.

(Example 6)
The removal material H was subjected to the same operation as in Example 4, and the average open area, open area ratio, and aspect ratio of the open area were measured. Table 3 shows the results.

(Comparative Example 7)
The removal material A was subjected to the same operation as in Example 4, and the average open area, open area ratio, and aspect ratio of the open area were measured. Table 3 shows the results.

(Comparative Example 8)
The removal material B was subjected to the same operation as in Example 4, and the average open area, open area ratio, and aspect ratio of open areas were measured. Table 3 shows the results.

(Comparative Example 9)
The removal material C was subjected to the same operations as in Example 4, and the average open area, open area ratio, and aspect ratio of open areas were measured. Table 3 shows the results.

(Comparative Example 10)
The removal material D was subjected to the same operation as in Example 4, and the average open area, open area ratio, and aspect ratio of the open area were measured. Table 3 shows the results.

(Comparative Example 11)
The removal material E was subjected to the same operation as in Example 4, and the average open area, open area ratio, and aspect ratio of the open area were measured. Table 3 shows the results.

(Comparative Example 12)
The removal material I was subjected to the same operation as in Example 4, and the average open area, open area ratio, and aspect ratio of open areas were measured. Table 3 shows the results.

From the results in Tables 2 and 3, the removal material with the average pore area per stitch limited to a specific range not only has a high removal rate of activated granulocyte-activated platelet complexes, but also activates It was also found that the ratio of clearance of granulocyte-activated platelet complexes and lymphocytes was increased.

Since the material for removing activated granulocyte-activated platelet complexes of the present invention can be suitably used for removing activated granulocyte-activated platelet complexes, it is useful for treating inflammatory diseases, particularly ARDS. It can be used as an extracorporeal circulation column.

1 hole part 1
2 Aperture Part 2
3 Fiber 4 Opening 5 Warp 6 Weft 7 Opening 8 Length of opening in wale direction 9 Length of opening in course direction

Claims (6)

comprising a knitted or woven fabric comprising fibers having ligands containing amide groups,
A material for removing activated granulocyte-activated platelet complexes, wherein the average open area per stitch of the knitted fabric or one stitch of the fabric is 0.080 mm ² or more and 0.100 mm ² or less. The knitted fabric is a weft knitted fabric,
The removal material according to claim 1, wherein the aspect ratio of the openings in the stitch is 0.75 to 1.30. 3. Removal material according to claim 1 or 2, wherein the fibers comprise polystyrene or polystyrene derivatives. The fiber is a sea-island composite fiber,
The sea component of the sea-island composite fiber is polystyrene or a polystyrene derivative,
The removal material according to any one of claims 1 to 3, wherein the island component of the sea-island composite fiber is polypropylene. The removal material according to any one of claims 1 to 4, wherein the average open pore area is 0.083 mm ² or more and 0.089 mm ² or less. A blood purifier comprising the removal material according to any one of claims 1-5.

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