JP2020092879A - Blood purification column - Google Patents

Abstract

To provide a blood purification column capable of suppressing an increase in pressure without deteriorating adsorption object substance adsorbing ability.SOLUTION: A blood purification column includes a fibrous adsorption carrier 5 arranged around a central pipe. When an end surface of the fibrous adsorption carrier in an axial direction in touch with the central pipe is represented as a line segment M, an end surface of the fibrous adsorption carrier in an axial direction that is in non-contact with the central pipe and in contact with or close to an inner wall of a container is represented as a line segment N, and a development length of the fibrous adsorption carrier is represented as a line segment K, and when a point where a line segment connecting the middle points m and n of the respective line segments M and N intersects with a line extending in an axial direction from the middle point k of the line segment K is represented as A, and a point where a distance equivalent to 1/10 times as long as the line segment connecting the middle points m and n of the respective line segments M and N is moved in an expansion direction from the middle point of the line segment M is represented as B, a value (A/B) obtained by dividing an amount of anticoagulant adsorption at point A by an amount of anticoagulant adsorption at point B is 0.5-1.0.SELECTED DRAWING: Figure 3

Description

The present invention relates to a blood purification column.

So far, various blood purification columns filled with adsorbent carriers and solutions such as physiological saline have been developed for the purpose of adsorbing and removing inflammatory proteins and activated blood cells in blood for patients with inflammatory diseases. There is.

When extracorporeal circulation is performed for a patient with an inflammatory disease using a blood purification column, there is priming using a solution containing an anticoagulant as a preparatory step. There are the following two reasons for performing priming before using the column. The first is to discharge fine particles or suspended matter that may remain in the blood purification column, acidic or basic eluate, etc., and the second is to fill the column with an anticoagulant, By preventing the coagulation of blood components, which is one of the causes of blood clots and thrombus, it is possible to suppress the increase in circulation pressure when using a blood purification column and suppress the decrease in the adsorption rate of the adsorption target substance due to the loss of adsorption sites. This is because. Therefore, it is a well known fact that priming is essential for using a blood purification column. However, there has been no report on a blood purification column previously filled with an anticoagulant.

Examples of the liquid that is filled in the blood purification column by priming include a solution in which an anticoagulant or the like is dissolved. Blood purification columns before priming include those in a dry state (hereinafter, dry type) and those in a wet state (hereinafter, wet type). Particularly, a wet column includes water containing no salt as a packing liquid (hereinafter, It is generally filled with saline) or physiological saline. However, if extracorporeal circulation is carried out without priming with unsalted water in the column or with a filling solution having a pH changed by the acidic or basic eluate, there is a concern that blood coagulation is promoted.

Due to various time constraints, the priming time before use of the blood purification column is set to about 5 minutes in the clinical field, particularly in the field of emergency. At the priming time, liquid replacement of the aqueous solution containing the anticoagulant in the blood purification column is performed. As a result, insufficient adsorption of the anticoagulant occurs, and in some cases the blood component that has passed through the blood purification column coagulates inside the column, and becomes a liquid inlet, liquid outlet, adsorption carrier void, etc. Blood clots may become clogged. When the circulating pressure of the blood purification column suddenly rises, the extracorporeal circulation must be stopped, and there is a risk that patients who are in a hurry cannot receive prompt treatment.

Patent Document 1 discloses a method of immobilizing heparin, which is a kind of anticoagulant, on the surface of an adsorption carrier through an amino group and removing an etiological agent by utilizing electrostatic interaction of the amino group. .. It is disclosed that a circular membrane-like or non-woven fabric-like carrier, which is filled in a filter holder and has a diameter of about 25 mm, is suitable for removing HIV and endotoxin without inducing blood coagulation.

Patent Document 2 discloses a method of suppressing activation of blood platelets and blood coagulation by coating a fat-soluble antioxidant on the potting portion of a container filled with hollow fibers. It is disclosed that it is preferable that the agent is uniformly present on the end surface of the potting portion.

Patent Document 3 discloses a method of suppressing retention of blood in the column by paying attention to the arrangement of the holes in a column provided with a central pipe having holes on the side surface, and the distance between the holes is appropriately set. Is disclosed to be suitable.

Patent Document 4 discloses a fiber containing an amide group and an amino group. It is disclosed that a knitted fabric having a porosity of 0.1 to 30% is suitable for suppressing a pressure increase when passing blood.

JP, 11-267421, A JP, 2018-171431, A JP, 2017-170140, A International Publication No. 2018/047929

When heparin is present only in the vicinity of the adsorption carrier as in the column used in the example of Patent Document 1, blood coagulation occurs in a space other than the vicinity of the carrier when the inner volume of the column container increases. I have a concern. Further, when the column of Patent Document 1 is used in a wet state, heparin on the surface of the carrier is gradually released into the filling liquid, and there is a concern that sufficient suppression of blood coagulation cannot be exhibited.

In the hollow fiber blood purifier described in the example of Patent Document 2, the fat-soluble antioxidant is coated on the potting portion on the blood inlet or outlet side, but the entire carrier is not uniformly coated. Therefore, there is a concern that blood coagulation will occur when blood retention occurs in a portion where the antioxidant coating amount is small, and it is difficult to completely suppress blood coagulation inside the column with only the above antioxidant. It is supposed to be.

Patent Document 3 describes the relationship between the arrangement of the holes in the central pipe and the retention of blood, but does not describe the amount of anticoagulant adsorbed on the carrier or priming.

Patent Document 4 describes the relationship between the porosity of the knitted fabric and the pressure increase during blood flow, but does not describe the amount of anticoagulant adsorbed on the carrier or priming.

In order to suppress the pressure rise when using the blood purification column, it is necessary to reduce the adsorption unevenness of the anticoagulant during priming, and for that purpose it is considered that the blood purification column should be pre-filled with the anticoagulant. .. In that case, from the viewpoint of suppressing blood coagulation on the surface of the adsorption carrier, it is desirable that the adsorption carrier also contains a structure or a functional group capable of immobilizing or adsorbing the anticoagulant. However, since it is necessary to achieve both chemical stability that does not affect the pH of the filling liquid and adsorption ability of the substance to be adsorbed, it is considered that such a product has not been developed so far.

Therefore, it is an object of the present invention to provide a blood purification column that can achieve suppression of pressure increase without lowering the adsorption ability of a substance to be adsorbed.

As a result of intensive studies to solve the above-mentioned problems, the present inventors have performed a treatment for adsorbing an anticoagulant on a fibrous adsorption carrier in advance and filling the column, and then an aqueous solution containing an anticoagulant. The blood purification column filled with in the container, without decreasing the adsorption ability of the adsorption target substance to the fibrous adsorption carrier, adhesion of blood clots or thrombus to the adsorption carrier due to activation of blood components, the column It was found that the circulation pressure rise can be suppressed.

That is, the present invention includes the following (1) to (6).
(1) A container having a blood inlet and a blood outlet,
Inside the container, a central pipe and a fibrous adsorption carrier having a ligand containing an acidic functional group or a basic functional group bonded to the surface thereof are provided.
The container contains an aqueous solution containing an anticoagulant,
The central pipe has a plurality of holes on the side surface,
The fibrous adsorption carrier is arranged around the central pipe,
The average adsorption amount of the anticoagulant adsorbed on the fibrous adsorption carrier is 60 to 220 IU per 1 g of the dry weight of the fibrous adsorption carrier,
An end face of the fibrous adsorption carrier in the axial direction that comes into contact with the central pipe is defined as a line segment M, and an end face of the fibrous adsorption carrier in the axial direction that is not in contact with the central pipe but is in contact with or close to the inner wall of the container is defined. In the case where the line segment is N and the developed length of the fibrous adsorption carrier is the line segment K, from the line segment connecting the midpoints of the line segment M and the line segment N and the midpoint of the line segment K, The point where the line extending in the axial direction intersects is A, and the distance corresponding to 1/10 times the line segment connecting the midpoints of the line segment M and the line segment N is from the midpoint of the line segment M. A value (A/B) obtained by dividing the adsorption amount of the anticoagulant at the point A by the adsorption amount of the anticoagulant at the point B is 0.5 to 1.0, where B is the point moved in the developing direction. , Blood purification column.
(2) The blood volume of the container is 50 to 220 cm ³ ,
The blood purification column according to (1), wherein the fibrous adsorption carrier has a packing density of 0.2 to 0.5 g/cm ³ .
(3) The blood purification column according to (1) or (2), wherein the concentration of the anticoagulant is 2 to 40 IU/cm ³ .
(4) The blood purification column according to any one of (1) to (3), wherein the fibrous adsorption carrier is a knitted fabric or a woven fabric.
(5) The blood purification column according to any one of (1) to (4), wherein the anticoagulant is heparin, nafamostat mesylate or low molecular weight heparin.
(6) The fiber constituting the fibrous adsorption carrier is a sea-island composite fiber, the sea component is selected from the group consisting of polystyrene and its derivatives, polysulfone and its derivatives, and mixtures thereof, and the island component is polypropylene. The blood purification column according to any one of (1) to (5), which is selected from the group consisting of, polyethylene, polypropylene/polyethylene copolymer, and a mixture thereof.

The blood purification column of the present invention has a high adsorption ability for a substance to be adsorbed, can suppress the adhesion of blood clots or thrombi, and can suppress the pressure increase of the column.

It is a longitudinal cross-sectional view of an example of a radial flow type blood purification column. It is a figure showing a cylindrical fibrous adsorption carrier wound around a central pipe packed in an example of a radial flow type blood purification column. FIG. 3 is a view showing an expanded fibrous adsorption carrier wound around a central pipe after disassembling an example of a radial flow type blood purification column.

Hereinafter, the present invention will be described in detail.

The blood purification column of the present invention comprises a container having a blood inlet and a blood outlet, a central pipe inside the container, and a fibrous adsorption carrier having a ligand having an acidic functional group or a basic functional group bound on the surface thereof. And, the container contains an aqueous solution containing an anticoagulant therein, the central pipe has a plurality of holes on the side surface, the fibrous adsorption carrier is arranged around the central pipe, The average adsorption amount of the anticoagulant adsorbed on the fibrous adsorption carrier is 60 to 220 IU per 1 g of the dry weight of the fibrous adsorption carrier, and the end face of the fibrous adsorption carrier in the axial direction in contact with the central pipe is A line segment M is defined as an end face of the fibrous adsorption carrier in the axial direction which is in contact with or close to the inner wall of the container without contacting the center pipe, and a development segment of the fibrous adsorption carrier is defined as a line segment N. In the case of K, the line segment connecting the midpoints of the line segment M and the line segment N and the line extending from the midpoint of the line segment K in the axial direction are referred to as A, and the line Assuming that a point equivalent to 1/10 times the line segment connecting the midpoints of the segment M and the line segment N in the developing direction from the midpoint of the line segment M is B, A value (A/B) obtained by dividing the anticoagulant adsorption amount by the anticoagulant adsorption amount at the point B is 0.5 to 1.0.

The “adsorption carrier” means a carrier having an ability to adsorb an organic substance, and the presence or absence of the adsorption ability of other substances is not particularly limited as long as it has the ability to adsorb an organic substance.

"Adsorption" means a state in which a specific substance is attached to a material and is not easily separated, or an adsorption equilibrium state. Although the principle of adsorption is not particularly limited, for example, electrostatic interaction, hydrophobic interaction, hydrogen bond, adhesion state by intermolecular force such as Van der Waals force, cell adhesion, phagocytosis of leukocytes, etc. It means a state of being adhered.

“Blood component” means a component that constitutes blood, and examples thereof include humoral factors in blood and cells in blood. The blood component to be adsorbed by the blood purification column according to the present embodiment is not particularly limited, but among the blood components, the liquid factor in blood is suitable as the adsorption target substance.

The “liquid factor in blood” refers to an organic substance dissolved in blood. Specific examples include proteins such as urea, β2-microglobulin, cytokines, IgE and IgG, and polysaccharides such as lipopolysaccharide (LPS). Among them, proteins such as urea and cytokines and polysaccharides such as LPS are preferable as the adsorption target substance, and further, cytokines are more preferable as the adsorption target substance when the purpose is to treat inflammatory diseases.

"Cytokine" means a group of proteins that are produced by various cells including immunocompetent cells and are released extracellularly by the action of stimulation such as infection and trauma, and include, for example, interferon α, interferon β, Examples include interferon γ, interleukin 1 to interleukin 15, tumor necrosis factor-α, tumor necrosis factor-β, high mobility group box-1, erythropoietin or monocyte chemoattractant.

The "cells in blood" means cells contained in blood, and examples thereof include leukocyte components such as granulocytes, monocytes, neutrophils and eosinophils, erythrocytes and platelets. For the purpose of treating inflammatory diseases, it is desirable to adsorb and remove leukocyte component, activated leukocyte or activated leukocyte-activated platelet complex in addition to the humoral factor in blood. There is.

"Activated leukocytes" means leukocytes that release cytokines or active oxygen by stimulation with cytokines or LPS, and examples thereof include activated granulocytes and activated monocytes. The degree of activation of activated leukocytes can be determined by measuring the amount of activated oxygen or measuring the expression of surface antigens by flow cytometry or the like.

“Activated platelets” means platelets that release cytokines or active oxygen by stimulation with cytokines or LPS.

The term "activated leukocyte-activated platelet complex" means that the type of leukocyte is particularly limited as long as it is a complex in which activated leukocytes and activated platelets bind to each other, have a phagocytosis action on self-tissues, and release cytokines. It is not limited, and examples thereof include activated granulocyte-activated platelet complex and activated monocyte-activated platelet complex. In particular, in the treatment of patients with inflammatory diseases, it is considered important to remove the activated granulocyte-activated platelet complex that is considered to be directly involved in the pathological condition.

“Inflammatory disease” refers to all diseases in which an inflammatory reaction is induced in the body, and includes, for example, systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, ulcerative colitis, Crohn's disease, drug-induced hepatitis, alcoholic Hepatitis, hepatitis A, hepatitis B, hepatitis C, hepatitis D or hepatitis E, sepsis (eg, sepsis derived from Gram-negative bacteria, sepsis derived from Gram-positive bacteria, culture-negative sepsis, fungal sepsis), influenza , Acute respiratory distress syndrome (ARDS, also referred to as acute respiratory distress syndrome, acute respiratory distress syndrome), acute lung injury (ALI), pancreatitis, idiopathic interstitial pneumonia (Idiopathic) Pulmonary Fibrosis (IPF), inflammatory bowel disease (eg, ulcerative colitis, Crohn's disease), blood transfusion, organ transplantation, reperfusion injury after organ transplantation, cholecystitis, cholangitis, neonatal blood group incompatibility, etc. Be done. Among inflammatory diseases, drug-causing hepatitis, alcoholic hepatitis, hepatitis A, hepatitis B, hepatitis C, hepatitis D or E, in which causative substances are released into the blood and a therapeutic effect by blood purification can be expected particularly Hepatitis C, sepsis (eg, sepsis from Gram-negative bacteria, sepsis from Gram-positive bacteria, culture-negative sepsis, fungal sepsis), influenza, acute respiratory distress syndrome, acute lung injury, pancreatitis, idiopathic interstitial pneumonia, Is mentioned. As the use of the adsorption column of the present embodiment, for example, the above-mentioned use for treating inflammatory diseases is preferable, and among them, it is difficult to treat with a drug alone, and both cytokines and activated leukocytes-activated platelets are involved. Disease (eg, sepsis derived from Gram-negative bacteria, sepsis derived from Gram-positive bacteria, culture-negative sepsis, fungal sepsis), influenza, acute respiratory distress syndrome, acute lung injury, idiopathic interstitial pneumonia The therapeutic use of is more preferable.

The “anticoagulant” means a compound having a property of inhibiting blood coagulation that progresses due to activation of a blood coagulation factor typified by thrombin. Examples of the anticoagulant include aspirin, clopidogrel sulfate, prasugrel sulfate, ticlopidine hydrochloride, dipyridamole, cilostazol, beraprost sodium, limaprost alfadex, ozagrel sodium hydrochloride, sarpogrelate hydrochloride, ethyl icosapentate, trapidil, warfarin potassium, dalteparin sodium, Parnaparin sodium, levapirin sodium, rivaroxaban, apixaban, endoxaban, dabigatran, argatroban, heparin (eg, heparin sodium, heparin potassium, heparin calcium), low molecular weight heparin, heparin derivatives such as acetylated heparin, dextran sulfate, quenching Acid, nafamostat mesylate, polyvinyl sulfonic acid, polystyrene sulfonic acid and the like can be mentioned. Among them, when packed in a blood purification column or added directly to blood, from the viewpoint of safety to the living body, heparin, nafamostat mesylate, low molecular weight heparin, heparin derivatives such as acetylated heparin, dextran sulfate, polyvinyl sulfone. Acid, polystyrene sulfonic acid and the like are preferable, and heparin, nafamostat mesylate or low molecular weight heparin, which have many clinical results in extracorporeal circulation, are more preferable. Further, heparin, nafamostat mesylate or low molecular weight heparin may or may not be purified, and may form a salt with sodium, potassium or the like, which can inhibit blood coagulation reaction. If it is, it will not be specifically limited.

The “aqueous solution containing an anticoagulant” may be an aqueous solution prepared by dissolving an anticoagulant in water or a liquid prepared by adding an anticoagulant to the following aqueous solution and dissolving the solution. The liquid before dissolving the anticoagulant is not particularly limited, and examples thereof include distilled water, salt water, other buffer solutions, and deionized water. A preferred aqueous solution is an aqueous solution containing an inorganic salt, and physiological saline is more preferred. The inorganic salt is not particularly limited, and examples thereof include sodium chloride, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, or the corresponding potassium salt of the same acid, and a phosphate buffered physiological mixture of these. Saline solution is mentioned as an example. Since these components form a buffer containing the acid and its conjugate base, the addition of acid or base causes only a relatively small change in the pH of the liquid. The buffer may further be, for example, sodium borate, 2-(N-morpholino)ethanesulfonic acid (MES), sodium hydroxide, 2,2-bis(hydroxymethyl)-2,2′,2″-nitrilotriethanol. , N-tris(hydroxymethyl)methyl-2-aminoethanesulfonic acid, citric acid, sodium citrate, sodium carbonate, sodium hydrogen carbonate, acetic acid, sodium acetate, ethylenediaminetetraacetic acid or a combination thereof. .. The aqueous solution containing the anticoagulant is filled inside the container.

The concentration of the anticoagulant agent in the aqueous solution containing previous anticoagulant to be filled in the column is preferably from ^{1.5~40IU / cm 3, 2~40IU / cm} 3 is more preferable. By setting the concentration of the anticoagulant to 1.5 IU/cm ³ or more, it is possible to further suppress the risk that blood coagulates in the column or the blood circuit when the blood flows into the column and the circulation pressure increases. In addition, by setting the concentration of the anticoagulant to 40 IU/cm ³ or less, the concentration of the anticoagulant in the blood increases, it becomes difficult to stop the bleeding at the time of bleeding, and the risk of causing serious bleeding to the patient is further suppressed. be able to.

The “blood purification column” means a container having at least a blood inlet and a blood outlet. Examples of the container shape of the column include a radial flow type column. The container shape of the blood purification column may be a container having a lid member and a case portion in addition to the blood inlet port and the blood outlet port, as long as the adsorption carrier can be filled in the case portion.

Hereinafter, the blood purification column according to the present embodiment will be described in detail with reference to the drawings, but the present invention is not limited to these aspects. Also, the proportions in the drawings do not necessarily match those in the description.

The center pipe 4 shown in FIG. 1 is provided with a plurality of holes 9 provided on the side surface in the longitudinal direction of the center pipe 4 for allowing the supplied liquid to flow out. The number of the holes is not limited and may be appropriately set depending on the embodiment. Further, as the material of the central pipe, any material may be used as long as it is inert to the solvent used and the substance to be adsorbed and removed.

The fibrous adsorption carrier 5 is a carrier filled around the central pipe 4. The liquid supplied to the blood purification column 1 flows from the blood introduction port 2 of the lid material 11 to the central pipe 4, and flows out to the fibrous adsorption carrier 5 through the hole 9. When the liquid passes through the adsorption carrier 5, the adsorption target substance contained in the liquid is adsorbed, and the liquid after the adsorption treatment is discharged from the blood discharge port 3 of the lid material 12 to the outside of the column. In FIG. 1, the fibrous adsorption carrier 5 is filled as a cylindrical carrier around the center pipe 4 by winding a sheet-shaped knitted fabric around the center pipe 4. The shape of the fibrous adsorption carrier 5 and the method of filling it in the column are not limited to the above-mentioned methods, and any shape that can be placed inside the container 8 may be used. The thing of the shape etc. are mentioned. The material of the fibrous adsorption carrier 5 is appropriately selected according to the type of the substance to be adsorbed.

The plate 6 is a plate-shaped member for preventing the liquid from coming into contact with the fibrous adsorption carrier 5 without passing through the central pipe 4. The plate 6 is arranged so that the liquid that has flowed into the blood purification column 1 is communicated with the upstream end of the central pipe 4 so as to pass through the inner cavity of the central pipe 4. The plate 7 is a plate-shaped member for preventing the liquid flowing into the blood purification column 1 from flowing out of the column without passing through the hole 9 though passing through the central pipe 4. The plate 7 is arranged at the downstream end of the central pipe 4 so as to close the downstream end of the central pipe 4 and fix the fibrous adsorption carrier 5 in the space around the central pipe 4. Here, the shapes of the plate 6 and the plate 7 are not particularly limited, and any shape that can be arranged inside the container 8 may be used. The plate 6 and the plate 7 may be made of any material as long as it is inert to the solvent used and the substance to be adsorbed/removed. The shape of the container may be a prismatic container such as a cylinder, a triangular prism, a quadrangular prism, a hexagonal prism or an octagonal prism, but is not limited to this structure.

Examples of the material of the container 8 of the blood purification column 1 include glass, plastic/resin, stainless steel, etc., but there is no particular limitation, but operability at the clinical site or measurement site and ease of disposal are required. Considering this, the material is particularly preferably plastic or resin.

The lid member 11 and the lid member 12 are members that have a blood inlet or a blood outlet and that seal the fibrous adsorption carrier 5 and the aqueous solution 10 containing the anticoagulant filled in the container 8. Examples of the material of the lid material 11 and the lid seat 12 include those made of glass, plastic/resin, stainless steel, etc., but are not particularly limited, but from the viewpoint of good adhesiveness, they are the same materials as the column container. Preferably.

The blood inlet 2 and the blood outlet 3 can be replaced with each other depending on the direction in which the liquid flows.

The center pipe 4 and the fibrous adsorption carrier 5 shown in FIG. 2 are in a form for filling the container 8, and show a state in which the sheet-shaped fibrous adsorption carrier 5 is wound around the center pipe 4 in a cylindrical shape. The fibrous adsorption carrier 5 to be wound may be cut in advance in the axial length of the center pipe 4 and wound, or after winding, the fibrous adsorption carrier 5 having an extra length may be cut from the center pipe 4. Good.

The fibrous adsorption carrier 5 shown in FIG. 3 is a view in which the blood purification column 1 is disassembled and the adsorption carrier 5 wound around the central pipe 4 shown in FIG. 2 is developed. In FIG. 3, the length in the developing direction of the fibrous adsorption carrier 5 is defined as a line segment K, and when the adsorption carrier 5 has a cylindrical shape, the end face of the carrier in contact with the central pipe 4 in the axial direction is shown. A line segment M is defined as an end face of the carrier in the axial direction that is in non-contact with the central pipe 4 and is in contact with or close to the inner wall of the container 8, and the midpoint of the line segment K is k. When the middle point is m and the middle point of the line segment N is n, the point where the line segment connecting the middle points m and n and the line extending in the axial direction from the middle point k intersect is A. , B is a point corresponding to 1/10 of the line segment connecting the midpoint m and the midpoint n in the developing direction from the midpoint m, and 1/the line segment connecting the midpoint n and the midpoint m. Let C be a point that has moved a distance corresponding to 10 times from the midpoint n in the developing direction.

Hereinafter, the adsorption amount of the anticoagulant will be described by taking heparin as an example, but the anticoagulant of the present invention is not limited to heparin.

The "anticoagulant adsorption amount A" means the anticoagulant adsorption amount of the fibrous adsorption carrier at the intersection A shown in FIG. To measure the anticoagulant adsorption amount A, three fibrous adsorption carriers at the intersection point A were cut into a circle of Φ8 mm and reacted according to the operation procedure described in the package insert of Test Team Heparin S (Sekisui Medical Co., Ltd.), and the absorbance at 405 nm was measured. It can be determined by measuring with N=3 by a microplate reader (Corona Electric Co., Ltd., MTP-300). The “anticoagulant adsorption amount B” and the “anticoagulant adsorption amount C” can also be obtained by the same method as the anticoagulant adsorption amount A using each fibrous adsorption carrier at the intersection B and the intersection C.

The "average anticoagulant adsorption amount" is obtained by calculating the average value of the sum of the anticoagulant adsorption amounts on the respective fibrous adsorption carriers at the intersection points A, B and C obtained by the above method. , Can be calculated by the following equation 1.

Anticoagulant average adsorption amount (IU/g)=[{anticoagulant adsorption amount A(IU/g)}+{anticoagulant adsorption amount B(IU/g)}+{anticoagulant adsorption amount C(IU /G)}]/3...Equation 1

The average adsorption amount of the anticoagulant is set to 60 IU/g or more from the viewpoint of suppressing the coagulation of blood in contact with the fibrous adsorption carrier, whereby the anticoagulant is gradually released from the fibrous adsorption carrier during blood flow. It is difficult and sufficient suppression of blood coagulation can be achieved. Further, by setting it to 220 IU/g or less, it is possible to suppress the excessive adsorption of the anticoagulant on the adsorption carrier, and reduce the concern that the adsorption ability for the adsorption target substance such as cytokine may be reduced. That is, the average amount of the anticoagulant adsorbed needs to be 60 to 220 IU per 1 g of the dry weight of the fibrous adsorption carrier, preferably 80 to 150 IU, more preferably 80 to 100 IU.

The “value obtained by dividing A by B (A/B)” is the value obtained by dividing the anticoagulant adsorption amount at the intersection A calculated by the above method by the anticoagulant adsorption amount at the intersection B calculated by the above method. This means that it is an index showing unevenness of the anticoagulant adsorption amount in the whole fibrous adsorption carrier. The closer the value of A/B is to 1.0, the amount of anticoagulant adsorbed in the vicinity of the center pipe of the fibrous adsorption carrier wound in a cylindrical shape on the central pipe and the anticoagulation inside the adsorption carrier wound in a cylindrical shape. This indicates that there is no difference in the amount of adsorbed agent, and it is possible to suppress the generation of a blood clot or thrombus inside the cylindrical fibrous adsorption carrier. When the value of A/B is low, it means that the amount of the anticoagulant adsorbed inside the cylindrical adsorption carrier is low, and the blood flowing into the blood inlet and the center pipe contact portion passes through the fibrous adsorption carrier. At this time, there is a concern that a blood clot or a blood clot may be generated at the time when it reaches the inside of the cylindrical adsorption carrier. Therefore, the value of A/B needs to be 0.5 to 1.0, preferably 0.6 to 1.0, more preferably 0.7 to 1.0, and 0.8 to 1.0. It is more preferable that there is. In order to bring the value of A/B closer to 1.0, the sheet-shaped adsorption carrier is impregnated with an aqueous solution containing an anticoagulant before the column is filled with the fibrous adsorption carrier, and the entire adsorption carrier is This is achieved by uniformly adsorbing the anticoagulant on the surface. Further, by controlling the content of the acidic functional group or the basic functional group contained in the fibrous adsorption carrier within the range of 0.8 to 1.5 mmol per 1 g of the dry weight of the fibrous adsorption carrier, the value of A/B is further increased. Can be brought closer to 1.0.

When the packing density of the fibrous adsorption carrier in the blood purification column of the present embodiment is 0.2 g/cm ³ or more, the adsorption ability of the substance to be adsorbed can be more exerted, and is 0.5 g/cm ³ or less. ensuring blood volume by, since it can be filled with an aqueous solution containing a sufficient amount of anticoagulant is preferably 0.2-0.5 g / cm ^3, is 0.2-0.4 g / cm ³ More preferable.

The “packing density” is the dry weight (g) of the packed fibrous adsorption carrier per unit internal volume (cm ³ ) before packing the fibrous adsorption carrier inside the blood purification column, for example, the content of the column. When a container having a volume of 1 cm ³ is filled with 1 g of dry weight of the fibrous adsorption carrier, the packing density is 1 g÷1 cm ³ =1 g/cm ³ .

The “blood volume” can be calculated as a value obtained by subtracting the volume of the fibrous adsorption carrier in a wet state filled in the column container from the column internal volume. When the packing density is increased by increasing the filling amount of the fibrous adsorption carrier with respect to the inner volume of the column, the blood volume becomes relatively small. By setting the blood volume to 50 cm ³ or more, the blood flow path can be sufficiently secured, and by setting it to 220 cm ³ or less, the retention of blood and the filling of the fibrous adsorption carrier with the anticoagulant become uneven. The blood volume is preferably from 50 to 220 cm ³ , more preferably from 60 to 200 cm ³ , and even more preferably from 100 to 150 cm ³ , since it can be suppressed from occurring. Any of the preferred lower limits can be combined with any of the preferred upper limits.

The method for measuring the blood volume is shown below.
The blood purification column filled with the fibrous adsorption carrier is filled with water with the blood outlet blocked so that air does not enter. Next, it can be calculated by opening the blood outlet and taking out all the water filled in the column into a graduated cylinder and measuring the amount of the taken out water.

The method for measuring the dry weight of the fibrous adsorption carrier packed in the blood purification column is shown below.

First, a fibrous adsorption carrier having the same length and the same inner diameter as the space that can be filled with the adsorption carrier in the blood purification column container is prepared. When analyzing what has already been packed in the blood purification column, all the fibrous adsorption carrier in the column is taken out. After measuring the weight before drying, the fibrous adsorption carrier is allowed to stand for 8 hours in a vacuum state of 1 mmHg or less by a vacuum dryer set at 30°C. After standing still, the mass of the fibrous adsorption carrier is measured by an electronic balance. In order to determine that the adsorption carrier has dried, the difference in mass when the mass is measured hourly is 1 mass% or less as an index.

In the fibrous adsorption carrier, the cross-sectional shape of the fiber used is not particularly limited, and examples thereof include sea-island composite fibers having a sea-island cross section. The sea-island composite fibers include sea-island composite fibers alone and sea-island composite fibers having a suitable reinforcing material fixed or mixed. Here, the operation of fixing or mixing may be performed before or after processing the higher-order structure of the fiber.

There is no particular limitation on the component constituting the fibrous adsorption carrier, for example, when the shape of the fiber is a sea-island composite fiber, as the sea component, for example, polyethylene terephthalate, polybutylene terephthalate, polyaromatic vinyl compound, polyester, Mention may be made of polymers selected from the group consisting of polysulfones, polystyrenes and polyvinyl alcohols. The island component includes, for example, a polymer selected from the group consisting of polypropylene, polyethylene, an alloy or copolymer of polypropylene and polyethylene, and a mixture thereof. Regarding the sea component, from the viewpoint of improving the interaction with the organic substance to be adsorbed, polystyrene and its derivatives are obtained because the ligand containing an acidic functional group or a basic functional group is bonded to the surface of the fibrous adsorption carrier. , A polymer selected from the group consisting of polysulfones and their derivatives, and mixtures thereof, and the island component is selected from the group consisting of polypropylene, polyethylene, polypropylene/polyethylene copolymers and mixtures thereof. It is preferably a polymer.

The single yarn diameter of the fiber constituting the fibrous adsorption carrier may be any thickness, but from the viewpoint of improving the contact area with the substance to be adsorbed and maintaining the strength of the material, 3 to 200 μm is preferable, and 5 to 50 μm. Are more preferable, and 10-40 micrometers is still more preferable. Any of the preferred lower limits can be combined with any of the preferred upper limits.

The "single yarn diameter" means randomly picking 10 small fiber samples and taking 1000 to 3000 times photographs using a scanning electron microscope. Each of the 10 locations (total 100 locations) It means the average value of the measured values of the diameter of the fiber.

Examples of the shape of the fibrous adsorption carrier include knitted fabric, woven fabric, felt, and net, and knitted fabric or woven fabric is preferable. These can be manufactured by a known method using fibers as raw materials. For example, as a method for producing a felt, for example, a wet method, a carding method, an air-laying method, a spunbond method or a melt blow method can be mentioned, and as a method for producing a woven fabric, a plain weaving method, a jacquard weaving method can be mentioned, As a method for manufacturing the net, a circular knitting method or a cylinder knitting method can be mentioned. In particular, the knitted fabric is preferable from the viewpoint of filling a blood purifier with a large filling weight per unit volume.

Here, in the case of a blood purification column using a knitted fabric-shaped adsorption carrier, the porosity of the knitted fabric can be controlled by controlling the basis weight during knitting. By setting the porosity to 0.1% or more, it is possible to secure a sufficient blood flow path, and it is possible to further suppress an increase in circulation pressure due to clogging of the thrombus. Further, by setting the porosity to 30% or less, the adsorption performance for the target substance can be sufficiently exhibited. Therefore, the porosity of the knitted fabric-shaped adsorption carrier is preferably 0.1 to 30%, more preferably 1 to 30%, and particularly preferably 7 to 15%. Any of the preferred lower limits can be combined with any of the preferred upper limits.

"Pressure loss" means that when blood is passed through the blood inlet from the blood inlet in the axial direction of the blood purification column that uses a knitted fabric-shaped adsorption carrier, it passes through the blood purification column against the pressure applied to the blood. Represents the difference between the pressure applied to the blood and the pressure applied to the blood. Specifically, the blood inlet pressure and the blood outlet pressure when flowing blood through a blood purification column using a knitted fabric-shaped adsorption carrier are measured, respectively, and the value of the blood inlet pressure is used to determine the blood outlet pressure. The value obtained by subtracting the value is the pressure loss.

In the case of blood purification, if pressure loss occurs, there is a concern that the pressure will increase during blood purification. Therefore, the pressure at the inlet when the column is filled with the fibrous adsorption carrier and blood is passed at 100 cm ³ /min for 30 minutes. The pressure loss obtained by subtracting the value of the outlet pressure from the value of is preferably 50 mmHg or less, more preferably 30 mmHg or less, and particularly preferably 15 mmHg or less.

The “ligand” means a compound that binds to the surface of the fibrous adsorption carrier, and its chemical structure is not particularly limited as long as it contains an acidic functional group or a basic functional group. Examples thereof include a compound containing a sulfonic acid group or a carboxyl group which is (anionic functional group) or a compound containing an amino group which is a basic functional group (cationic functional group). In this embodiment, the ligand is preferably a compound containing a basic functional group, particularly a compound containing an amino group. The functional groups may be a combination of the same or different functional groups. The ligand may further have a neutral functional group as long as it has the acidic functional group or the basic functional group, and the neutral functional group is, for example, a methyl group or an ethyl group. Alkyl group or phenyl group, a phenyl group substituted with an alkyl group (for example, para(p)-methylphenyl group, meta(m)-methylphenyl group, ortho(o)-methylphenyl group, para(p)- Ethylphenyl group, meta(m)-ethylphenyl group, ortho(o)-ethylphenyl group, etc., or a phenyl group substituted with a halogen atom (for example, para(p)-fluorophenyl group, meta(m)-fluoro). An aryl group such as a phenyl group, an ortho(o)-fluorophenyl group, a para(p)-chlorophenyl group, a meta(m)-chlorophenyl group or an ortho(o)-chlorophenyl group is an acidic functional group or a base. A compound bound to a compound containing a functional group (eg, tetraethylenepentamine bound to a para(p)-chlorophenyl group) is included in the ligand. At that time, the neutral functional group and the ligand may be directly bound or may be bound via a spacer (the spacer involved in the binding is also referred to as spacer 1). Examples of the spacer 1 include a urea bond, an amide bond, and a urethane bond.

The “amino group” is, for example, an amino group derived from a primary amine such as methylamine, ethylamine, propylamine, butylamine, pentylamine, hexylamine, heptylamine, octylamine or dodecylamine, methylhexylamine, diphenylmethylamine. , Amino groups derived from secondary amines such as dimethylamine, amino groups derived from amines having unsaturated alkyl chains such as allylamine, amino groups derived from tertiary amines such as trimethylamine, triethylamine, dimethylethylamine, phenyldimethylamine and dimethylhexylamine. Group, amino group derived from amine having aromatic ring such as 1-(3-aminopropyl)imidazole, pyridin-2-amine, 3-sulfoaniline or tris(2-aminoethyl)amine, ethylenediamine, diethylenetriamine, triethylenetetramine , Tetraethylenepentamine, dipropylenetriamine, polyethyleneimine, N-methyl-2,2'-diaminodiethylamine, N-acetylethylenediamine, 1,2-bis(2-aminoethoxyethane), etc., alkyl chain, aromatic Examples thereof include amino groups derived from a compound (hereinafter referred to as “polyamine”) in which two or more amino groups are bonded with a compound, a heterocyclic compound, a monocyclic compound, or the like, but a polyamine-derived amino group is preferable. In particular, an amino group derived from ethylenediamine, diethylenetriamine, triethylenetetramine or tetraethylenepentamine is preferable, and an amino group derived from tetraethylenepentamine is more preferable. Further, the amino group is more preferably an amino group derived from a primary amine or a secondary amine.

In the present embodiment, the fibrous adsorption carrier and the ligand containing an acidic functional group or a basic functional group may be directly bonded, or the reactive functional group derived from the reactive functional group may be present between the fibrous adsorption carrier and the ligand. It may be via a spacer (the spacer involved in the binding is also referred to as spacer 2). The spacer 2 may be any as long as it has an electrically neutral chemical bond such as a urea bond, an amide bond, an ether bond, an ester bond, a urethane bond, or the like, and has an amide bond or a urea bond. Those that are present are preferable.

The reactive functional group that mediates the bond between the fibrous adsorption carrier and the ligand, for example, an active halogen group such as a halomethyl group, a haloacetyl group, a haloacetamidomethyl group or a halogenated alkyl group, an epoxide group, a carboxyl group, Examples thereof include an isocyanic acid group, a thioisocyanic acid group, and an acid anhydride group. From the viewpoint of having suitable reactivity, an active halogen group is preferable, and a haloacetamidomethyl group is more preferable. Specific examples of the polymer material having a reactive functional group introduced therein include polystyrene having a chloracetamidomethyl group added thereto and polysulfone having a chloracetamidomethyl group added thereto.

The reactive functional group can be bound to the fibrous adsorption carrier by previously reacting the fibrous adsorption carrier with an appropriate reagent. For example, when the component constituting the fibrous adsorption carrier is polystyrene and the reactive functional group is a chloroacetamidomethyl group, the chloroacetamidomethyl group is bonded by reacting polystyrene with N-hydroxymethyl-2-chloroacetamide. Polystyrene can be obtained. By reacting, for example, tetraethylenepentamine having an amino group with polystyrene having a chloroacetamidomethyl group bonded thereto, polystyrene having tetraethylenepentamine bonded via an acetamidomethyl group can be obtained. In this case, the acetamidomethyl group corresponds to spacer 2 and tetraethylenepentamine corresponds to the ligand. The materials of the sea component and the island component of the fibrous adsorption carrier, the spacer (spacer 1 and spacer 2), and the ligand can be arbitrarily combined. Examples of the constituent components of the fibrous adsorption carrier to which the ligand is bound, ethylenediamine, diethylenetriamine, compounds containing an amino group derived from triethylenetetramine or tetraethylenepentamine, polystyrene or ethylenediamine bound via an acetamidomethyl group, diethylenetriamine, Examples thereof include polysulfone in which a compound containing an amino group derived from triethylenetetramine or tetraethylenepentamine is bound via an acetamidomethyl group.

There is no particular limitation on the content of the acidic functional group or the basic functional group, but by setting the amount of 0.4 mmol or more per 1 g of the dry weight of the fibrous adsorption carrier, the adsorption performance for charged organic substances such as blood components is sufficiently improved. The pH of blood components can be prevented from changing by adjusting the amount to 2.0 mmol or less. Therefore, the content of the acidic functional group or the basic functional group is preferably 0.4 to 2.0 mmol, more preferably 0.8 to 1.5 mmol, per 1 g of the dry weight of the fibrous adsorption carrier. Any of the preferred lower limits can be combined with any of the preferred upper limits.

The content of the acidic functional group or the basic functional group can be measured by an acid-base titration method using a hydrochloric acid or sodium hydroxide aqueous solution.

In the fibrous adsorption carrier, since it is necessary to interact with the substance to be adsorbed, it is necessary that the ligand is bound to at least the surface side in contact with an organic substance such as blood. Here, the surface means the surface of the fibrous adsorption carrier, and in the case of a shape having pores on the surface, the outermost layer portion along the irregularities of the pores is also included in the surface. Further, when the fiber has through holes, not only the outermost layer portion of the fibrous adsorption carrier but also the outer layer of the through holes inside the adsorption carrier is included in the surface.

The fibrous adsorption carrier can be produced by the following method, but is not limited to this method.

The fiber diameter of the fibrous adsorption carrier can be reduced by reducing the polymer discharge amount during spinning and increasing the winding speed. When the ligand is introduced, the fiber diameter can be increased by swelling by impregnation with a solvent at the time of introducing the ligand. Therefore, the fiber diameter can be controlled within a target range by appropriately adjusting the conditions.

The shape having pores on the fiber surface can be controlled by impregnating the fiber with a solvent and manufacturing the fiber by etching. For example, when the fiber is a sea-island composite fiber, the pores can be increased by impregnation with a solvent in which the sea component is easily dissolved. Also, since it can be increased by adding a crosslinking agent and a catalyst to the mixed solution at the same time, the pore shape can be controlled by appropriately adjusting the conditions.

Examples of the solvent include nitrobenzene, nitropropane, chlorobenzene, toluene and xylene when the sea component is polystyrene, and nitrobenzene or nitropropane is preferable.

Examples of the cross-linking agent include aldehyde compounds such as paraformaldehyde, acetaldehyde or benzaldehyde.

As the catalyst for crosslinking, for example, sulfuric acid, hydrochloric acid, nitric acid or a Lewis acid such as aluminum (III) halide (eg, aluminum (III) chloride) or iron (III) halide (eg, iron (III) chloride) And sulfuric acid or iron (III) chloride is preferably mixed.

The concentration of the catalyst in the mixed liquid is preferably 5 to 80 wt%, more preferably 30 to 70 wt%.

The impregnation temperature is preferably 0 to 90°C, more preferably 5 to 40°C.

The impregnation time is preferably 1 minute to 120 hours, more preferably 5 minutes to 24 hours.

An example of the method for modifying the ligand on the fibrous adsorption carrier will be described below. A Lewis acid (eg, aluminum (III) chloride) and a carbamic acid chloride having a halogenated alkyl group (eg, N,N-bis(2-chloroethyl)carbamic acid chloride) were dissolved in a non-polar solvent (eg, dichloromethane). The sea-island composite fiber is added to the solution and stirred to form a carbamic acid chloride-bonded sea-island composite fiber. Then, the fiber is taken out, and subsequently, the carbamic acid chloride-bonded sea-island composite fiber is added to a dimethyl sulfoxide (hereinafter, DMSO) solution in which an amine compound (for example, tetraethylenepentamine) is dissolved as a ligand, and the water is taken out. The sea-island composite fiber having a compound containing an amino group as a ligand bonded to the surface is washed with.

As a method for producing a blood purification column according to the present embodiment, a container having a blood inlet and a blood outlet is filled with a fibrous adsorbent carrier wound around a central pipe and filled with an aqueous solution containing an anticoagulant. To be achieved. The following method can be given as a specific example of creation, but is not limited to the following form. The sheet-shaped fibrous adsorption carrier 5 is wound around the central pipe 4 so as to have a cylindrical shape. It is confirmed that the diameter of the wound cylindrical fibrous adsorption carrier 5 is not more than the inner diameter of the space in the cylindrical container 8 in which the adsorption carrier can be filled. Subsequently, the plate 6 and the plate 7 are fitted into both ends of the central pipe 4 around which the fibrous adsorption carrier 5 is wound, the containers 8 are filled with these, and the lid member 11 serving as a blood inlet and the lid member serving as a blood outlet. Attach 12. Finally, it can be prepared by filling the aqueous solution 10 containing the anticoagulant from the blood inlet 2.

The blood purification column of the present embodiment is intended to adsorb blood components, but in the case of inflammatory diseases, cytokine, which is a liquid factor in blood, is particularly preferably used as a substance to be adsorbed. Therefore, the blood purification column of the present embodiment can be used for extracorporeal circulation treatment of patients with inflammatory diseases.

As a method for evaluating the blood purification performance of the fibrous adsorption carrier, for example, a method of measuring an interleukin 8 (hereinafter, IL-8) adsorption rate can be mentioned. IL-8 is a kind of cytokine contained in blood components, and is known to have a markedly high value in blood components of patients caused by bronchiolitis and viral infection, especially in patients with inflammatory diseases. It is suitable as a blood component for blood purification performance evaluation. It can be judged that the higher the adsorption rate of IL-8, the higher the blood purification performance of the fibrous adsorption carrier.

Hereinafter, the blood purification column according to the present embodiment will be specifically described with reference to Examples and Comparative Examples, but the present invention is not limited to these examples.

(Preparation of fiber A)
Separately melted and measured using polystyrene as the sea component and polypropylene as the island component, and let them flow into a spinning pack incorporating a sea-island composite spinneret with 704 distribution holes for the island component per discharge hole. A composite stream was formed and melted and discharged. The island ratio was controlled to 50 wt%, the distance from the surface of the sea-island composite fiber to the outermost island component was adjusted to 2 μm, and the sea-island composite fiber (hereinafter, fiber A) having a single fineness of 3.0 dtex (fiber diameter 20 μm) was used. Obtained.

(Preparation of knitted fabric A)
Using the fiber A, a stitch adjusting scale of a tubular knitting machine (model name: circular knitting machine MR-1, Maruzen Sangyo Co., Ltd.) is adjusted to give a weight per unit area of 60 g/m ² and a bulk density of 0.20 g/cm ³ . A knitted fabric (hereinafter, knitted fabric A) was produced.

(Preparation of adsorption carrier 1)
N- hydroxymethyl-2-chloroacetamide (hereinafter, NMCA) after addition of 117g in a mixed solution of nitrobenzene 650 cm ³ and 98 wt% sulfuric acid 423cm ^3, and stirred at 10 ° C. until dissolution is NMCA, was prepared NMCA solution .. Next, nitrobenzene 50 cm ^3, 98 wt% paraformaldehyde in a mixed solution of sulfuric 33cm ³ (hereinafter, PFA) was added 5.0 g, was stirred at 20 ° C. until PFA is dissolved to prepare a PFA solution. The PFA solution (85 cm ³⁾ was cooled to 5° C. and then mixed with the NMCA solution (1100 cm ³ ). After stirring the mixed solution for 5 minutes, 25 g of the knitted fabric A was added and impregnated for 2 hours. After the impregnated knitted fabric A was immersed in 1100 cm ³ of nitrobenzene at 0° C. to stop the reaction, nitrobenzene attached to the knitted fabric A was washed with methanol.

Tetraethylenepentamine (hereinafter, TEPA) and 4.1 cm ³ of triethylamine 71cm ³ to the mixed solution obtained by dissolving the DMSO1000cm ^3, the knitted fabric A after washing with the above methanol added as it, 3 hours impregnated at 40 ° C. Let The knitted fabric A was filtered using a glass filter and washed with 1000 cm ³ of DMSO.

1.88 g of parachlorophenylisocyanate was added to 1200 cm ³ of DMSO dehydrated and dried with active molecular sieves 3A under a nitrogen atmosphere and heated to 30° C., and the entire amount of the knitted fabric A after the washing was impregnated for 1 hour. The knitted fabric A is filtered off using a glass filter to obtain a fibrous adsorption carrier 1 (unit weight: 100 g/m ² , thickness: 0.30 mm, open area ratio: 7.7%, hereinafter, adsorption carrier 1). It was

Measurement of the content of basic functional groups contained in the adsorption carrier 1:
The content of the basic functional group contained in the adsorption carrier 1 was determined by acid-base back titration of the basic functional group in the adsorption carrier 1. 1.5 g of the adsorption carrier 1 was placed in a 200 cm ³ eggplant flask and left to stand at 80° C. for 48 hours under normal pressure in a dryer to obtain a dried adsorption carrier 1. Next, 1.0 g of the above adsorption carrier 1 and 50 cm ³ of a 6M aqueous sodium hydroxide solution were added to a polypropylene container, stirred for 30 minutes, and the adsorption carrier 1 was filtered off using a filter paper. Next, the adsorption carrier 1 was added to 50 cm ³ of ion-exchanged water, stirred for 30 minutes, and separated by filtration using a filter paper. The adsorbent carrier 1 after desalting was obtained by repeating the operation of adding the adsorbent carrier 1 to ion-exchanged water, washing and filtering until the pH of the washing liquid after filtering the added ion-exchanged water reached 7. The desalted adsorption carrier 1 was allowed to stand for 8 hours under a vacuum condition with a vacuum dryer set at 30°C. Subsequently, 1.0 g of the adsorption carrier 1 and 30 cm ^{3 of} 0.1 M hydrochloric acid were added to a polypropylene container, and the mixture was stirred for 10 minutes. After stirring, 5 cm ^{3 of} the solution alone was extracted and transferred to a polypropylene container. Next, 0.1 cm ³ of 0.1 M sodium hydroxide aqueous solution was added dropwise to the extracted solution. After dropping, the solution was stirred for 10 minutes and the pH of the solution was measured. After the dropping of the 0.1 M sodium hydroxide aqueous solution, stirring for 10 minutes and pH measurement were repeated 100 times in the same manner. The amount of 0.1 M aqueous sodium hydroxide solution added when the pH of the solution exceeded 8.5 was defined as the titration amount per 1 g. The content of the basic functional group per 1 g of the adsorption carrier 1 was calculated using the titration amount per 1 g and the following formula 2. As a result, the basic functional group content of the adsorption carrier 1 was 1.0 mmol/g.

Content of basic functional group per 1 g of dry weight of the adsorption carrier 1 (mmol/g)={liquid amount of added 0.1M hydrochloric acid (30 cm ³ )/liquid amount of extracted hydrochloric acid (5 cm ³ )}×per 1 g Titration amount (cm ³ /g)×sodium hydroxide aqueous solution concentration (0.1 mol/L)...Equation 2

(Preparation of adsorption carrier 2)
A fiber having a basic functional group content of 0.2 mmol/g by performing the same operation and the same measurement as in the production method of the adsorption carrier 1 except that the addition amount of tetraethylenepentamine was changed to 1.0 cm ^3. -Like adsorption carrier 2 (basis weight: 95 g/m ² , thickness: 0.32 mm, open area ratio: 6.9%, hereinafter adsorption carrier 2) was obtained.

(Preparation of adsorption carrier 3)
Heparin sodium was dissolved in physiological saline to prepare 2 L of a 4 IU/cm ³ sodium heparin saline solution. Then, after the knitted fabric size was washed adsorption carrier 1 of 15cm × 280 cm of saline 2L, is immersed adsorption carrier 1 in sodium heparin saline solution 2000 cm ³ of 4 IU / cm ³ was prepared, 37 ° C. atmospheric It was left still for 30 minutes under pressure condition. After standing, the supernatant was removed to obtain a heparin-immobilized fibrous adsorption carrier 3 (Basis weight: 100 g/m ² , thickness: 0.30 mm, open area ratio: 7.7%, hereinafter, adsorption carrier 3). It was

(Preparation of adsorption carrier 4)
The fibrous adsorption carrier 4 (basis weight: 100 g/m ² , thickness: by performing the same operation as the method for producing the adsorption carrier 3 except that the concentration of the heparin sodium physiological saline solution to be dipped was changed to 2 IU/cm ^3. 0.30 mm, open area ratio: 7.7%, and the following adsorption carrier 4) was obtained.

(Preparation of adsorption carrier 5)
By performing the same operation as in the method for producing the adsorption carrier 3 except that the adsorption carrier 1 was changed to the adsorption carrier 2, the fibrous adsorption carrier 5 (area weight: 95 g/m ² , thickness: 0.32 mm, porosity: 6.9%, hereinafter, adsorption carrier 5) was obtained.

(Example 1)
A suction pipe 3 having a width of 12.4 cm, a length of 280 cm, and a thickness of 0.30 mm is wound around a central pipe having a diameter of 12.4 cm and a diameter of 1.2 cm and a plurality of holes on the side surface in the longitudinal direction. The flow column container was filled. Subsequently, a blood purification column 1 (blood volume: 135 cm ³ , hereafter, column 1) was prepared by filling 4 IU/cm ³ of sodium heparin saline solution so that the air could completely escape from the blood inlet of the column container. did. In addition, in order to carry out the following three tests, three identical columns were prepared and designated as Examples 1-1, 1-2, and 1-3, respectively. Similarly, in Examples 2 to 4 and Comparative Examples 1 to 3, three columns were prepared. Refer to FIG. 1 for other configurations of the column.

1. Measurement of heparin adsorption on column 1:
In order to measure the amount of heparin adsorbed on the adsorption carrier 3 packed in the obtained column 1, the column 1 was disassembled with a pipe cutter, and the adsorption carrier wound around the central pipe was taken out. Then, the adsorption carrier wound around the central pipe is unwound, and the adsorption carrier 3 at the intersections A, B, and C shown in FIG. 3 is cut into a circle of Φ8 mm, put into a plastic tube, and 2 cm ³ of physiological saline is added. After rinsing at room temperature, the reaction was performed according to the operating procedure of Test Team Heparin S (Sekisui Medical Co., Ltd.), and the absorbance at 405 nm was measured with a microplate reader (Corona Electric Co., Ltd., MTP-300) at N=3. According to the operating procedure of the kit, the values at the intersection points A and B and the average values of the three points of the intersection points A, B and C were calculated, and the heparin adsorption amount was obtained. The results are shown in Table 1 (Example 1-1).

2. Column 1 blood flow test:
Using the obtained column 1, pressure measurement was performed by a blood flow test using fresh bovine blood. Circuits were connected to the blood inlet and the blood outlet of the column 1, respectively, and pressure gauges (AP32A, manufactured by KEYENCE CORPORATION) were attached to the circuits immediately before the blood inlet and immediately after the blood outlet, and kept at 37°C (outside temperature). Fresh cow blood was passed at 100 cm ³ /min, and the pressures at the blood inlet and blood outlet were measured. A value (differential pressure) was calculated by subtracting the blood outlet pressure at 30 minutes after the start of the liquid passage from the blood inlet pressure at the time of 30 minutes after the start of the liquid passage. After confirming the pressure, the liquid flow was stopped, physiological saline was flowed at 100 cm ³ /min for 5 minutes to wash the inside of the column, and then the column was disassembled to determine whether or not a blood clot or thrombus was generated. The presence of blood clots or thrombus in the column or the adsorption carrier was judged to be "present", and if not observed, it was judged to be "none". The pressure measurement result and the blood clot or thrombus determination result are shown in Table 2 (Example 1-2).

3. Column 1 IL-8 adsorption rate measurement:
Using the obtained column 1, the adsorption rate of IL-8 was measured. Human IL-8 was prepared in fetal bovine serum to a concentration of 10 ng/cm ³ , and 2 L of the IL-8-containing solution kept at 37° C. (external temperature) was circulated in column 1 at 50 cm ³ /min. The IL-8 adsorption rate was calculated by the following equation 3 when the concentration of the IL-8-containing solution was C0 and the concentration of the IL-8-containing solution after 1-hour circulation was C1. The results are shown in Table 3 (Example 1-3).

IL-8 adsorption rate (%)=(C0−C1)/C0×100 Equation 3

(Example 2)
A blood purification column 2 (blood volume: 135 cm ³ , hereinafter, column 2) was produced by performing the same operation as in Example 1 using the adsorption carrier 4. The column 2 thus obtained is subjected to the same measurement as in Example 1 to determine the amount of heparin adsorbed (Example 2-1), the pressure during blood flow, and the determination of the adherence of blood clots or thrombi (Example 2-2). , IL-8 adsorption rate (Example 2-3) was measured. The results are shown in Table 1, Table 2 and Table 3.

(Example 3)
Same as Example 1 except that the concentration of the heparin sodium physiological saline solution to be filled was changed to 36 IU/cm ³ using the adsorption carrier 4 having a knitted fabric size of 12.4 cm in width, 175 cm in length, and 0.30 mm in thickness. The blood purification column 3 (blood volume: 200 cm ³ , hereinafter, column 3) was produced by performing the operation of. The obtained column 3 is subjected to the same measurement as in Example 1 to determine the amount of heparin adsorbed (Example 3-1), the pressure during blood flow, and the determination of blood clot or thrombus adhesion (Example 3-2). , IL-8 adsorption rate (Example 3-3) was measured. The results are shown in Table 1, Table 2 and Table 3.

(Example 4)
Same as Example 1 except that the concentration of the heparin sodium physiological saline solution to be filled was changed to 2 IU/cm ³ using the adsorption carrier 4 having a knitted fabric size of 12.4 cm in width, 370 cm in length, and 0.30 mm in thickness. The blood purification column 4 (blood volume: 100 cm ³ , hereinafter, column 4) was produced by performing the operation of. The obtained column 4 is subjected to the same measurement as in Example 1 to determine the amount of heparin adsorbed (Example 4-1), the pressure during blood flow, and the determination of the adherence of a blood clot or thrombus (Example 4-2). , IL-8 adsorption rate (Example 4-3) was measured. The results are shown in Table 1, Table 2 and Table 3.

(Comparative Example 1)
The adsorption carrier 1 having a width of 12.4 cm, a length of 280 cm, and a thickness of 0.30 mm is wound around a central pipe having a diameter of 12.4 cm and a diameter of 1.2 cm and a plurality of holes on the side surface in the longitudinal direction, and the radial is formed. The flow column container was filled. Subsequently, physiological saline was filled so that the air could completely escape from the blood inlet of the column container, to prepare a blood purification column 5 (blood volume: 135 cm ³ , hereinafter, column 5).

4. Measurement of heparin adsorption on column 5:
Using the obtained column 5, priming was carried out by passing 4 IU/cm ³ of heparin sodium physiological saline solution at 100 cm ³ /min for 10 minutes from the blood inlet, and then the heparin adsorption amount of the column 5 was measured. The result measured by the same operation as 1 is shown in Table 1 (Comparative Example 1-1).

5. Column 5 blood flow test:
Using the obtained column 5, pressure measurement was performed by a blood flow test using fresh bovine blood. Circuits were connected to the blood inlet and the blood outlet of the column 5, respectively, and priming was performed by passing 4 IU/cm ³ of sodium heparin saline solution at 100 cm ³ /min for 10 minutes from the blood inlet. Subsequently, a pressure gauge (AP32A, manufactured by KEYENCE CORPORATION) was attached to the circuit immediately before the blood inlet and immediately after the blood outlet, and fresh cow blood kept at 37°C (external temperature) was passed at 100 cm ³ /min to give blood. The pressures at the inlet and the blood outlet were measured respectively. A value was calculated by subtracting the blood outlet port pressure at 30 minutes after the start of liquid passage from the blood inlet port pressure at 30 minutes after the start of liquid passage. After confirming the pressure, the liquid flow was stopped, physiological saline was flowed at 100 cm ³ /min for 5 minutes to wash the inside of the column, and then the column was disassembled to determine whether or not a blood clot or thrombus was generated. The presence of blood clots or thrombus in the column or the adsorption carrier was judged to be "present", and if not observed, it was judged to be "none". The pressure measurement result and the blood clot or thrombus determination result are shown in Table 2 (Comparative Example 1-2).

6. Column 5 IL-8 adsorption rate measurement:
The results of measuring the IL-8 adsorption rate of column 5 by the same operation as in Example 1 are shown in Table 2 (Comparative Example 1-3).

(Comparative example 2)
A blood purification column 6 (blood volume: 135 cm ³ , hereinafter referred to as the column) was performed by performing the same operation as in Example 1 except that the adsorbing carrier 4 was used to change the packed sodium heparin physiological saline solution to physiological saline. 6) was produced. The obtained column 6 is subjected to the same measurement as in Example 1 to determine the amount of heparin adsorbed (Comparative Example 2-1), the pressure during blood flow, and the determination of blood clot or thrombus adhesion (Comparative Example 2-2). , IL-8 adsorption rate (Comparative Example 2-3) was measured. The results are shown in Table 1, Table 2 and Table 3.

(Comparative example 3)
By performing the same operation as in Example 1 using the adsorption carrier 5, a blood purification column 7 (blood volume: 135 cm ³ , hereinafter, column 7) was produced. The obtained column 7 is subjected to the same measurement as in Example 1 to determine the amount of heparin adsorbed (Comparative Example 3-1), the pressure at the time of blood passage, and the determination of blood clot or thrombus adhesion (Comparative Example 3-2). , IL-8 adsorption rate (Comparative Example 3-3) was measured. The results are shown in Table 1, Table 2 and Table 3.

In Table 1, the adsorption amount A means the heparin adsorption amount at the intersection point A shown in FIG. 3, the adsorption amount B means the heparin adsorption amount at the intersection point B shown in FIG. 3, and the heparin adsorption treatment “Yes” is the column. It means that the adsorption carrier that had been subjected to the heparin adsorption treatment was used before packing, and "no" for the heparin adsorption treatment means that the adsorption carrier that had not been subjected to the heparin adsorption treatment was used before the column packing, and the priming "with" ”Means that a column that has been subjected to a priming operation for passing an aqueous solution containing an anticoagulant before the blood passage test was used, and “no” priming means an aqueous solution containing an anticoagulant before the blood passage test. It means that a column which has not been subjected to the priming operation for passing the liquid was used.

From the results of Table 1, when only priming is performed using the fibrous adsorption carrier that has not been subjected to the heparin adsorption treatment, the heparin adsorption amount on the fibrous adsorption carrier becomes uneven, while the blood purification column according to the present embodiment Then, it was clarified that unevenness of the adsorption amount of heparin can be reduced by filling the fibrous adsorption carrier which has been subjected to the heparin adsorption treatment in advance.

From the results of Table 2, it is clear that by using a column filled with an heparin adsorption treatment and an aqueous solution containing heparin, it is possible to suppress an increase in pressure during permeation and prevent adhesion of a blood clot or a thrombus even in priming-free. It was Also, when performing priming for the purpose of washing impurities in the column, by using an aqueous solution containing heparin, it is possible to circulate blood in a state in which the unevenness of heparin adsorbed on the adsorption carrier is low.

From the results in Table 3, even when heparin was adsorbed on the fibrous adsorption carrier and the heparin-containing aqueous solution was packed in the blood purification column, it was possible to sufficiently adsorb the cytokine as the substance to be adsorbed. It is considered that the column can exhibit sufficient performance as a blood purification column.

The blood purification column of this embodiment can be used for biological component treatment in the medical field, particularly for blood component treatment.

1. Blood purification column 2. Blood inlet/blood outlet 3. Blood outlet/blood inlet 4. Central pipe 5. Fibrous adsorption carrier 6. Plate 7. Plate 8. Container 9. Hole 10. Aqueous solution containing anticoagulant 11. Lid 12. Lid material A. A point where a line segment connecting the midpoint m and the midpoint n intersects with a line extending from the midpoint k in the axial direction B. A point obtained by moving a distance corresponding to 1/10 times the line segment connecting the midpoint m and the midpoint n from the midpoint m in the developing direction. A point obtained by moving a distance corresponding to 1/10 times the line segment connecting the midpoint n and the midpoint m from the midpoint n in the developing direction K. A line segment M. representing the length of the fibrous adsorption carrier 5 in the developing direction. A line segment showing the end face in the axial direction that was in contact with the central pipe 4 N. A line segment showing an end face in the axial direction that is not in contact with the central pipe 4 and is in contact with or close to the inner wall inside the container 8 k. Midpoint of line segment m. Midpoint of line segment M n. Midpoint of line segment N

Claims (6)

A container having a blood inlet and a blood outlet,
Inside the container, a central pipe and a fibrous adsorption carrier having a ligand containing an acidic functional group or a basic functional group bonded to the surface thereof are provided.
The container contains an aqueous solution containing an anticoagulant,
The central pipe has a plurality of holes on the side surface,
The fibrous adsorption carrier is arranged around the central pipe,
The average adsorption amount of the anticoagulant adsorbed on the fibrous adsorption carrier is 60 to 220 IU per 1 g of the dry weight of the fibrous adsorption carrier,
An end face of the fibrous adsorption carrier in the axial direction that contacts the central pipe is defined as a line segment M, and an end face of the fibrous adsorption carrier in the axial direction that is not in contact with the central pipe and is in contact with or close to the inner wall of the container is defined. In the case where the line segment is N and the developed length of the fibrous adsorption carrier is the line segment K, from the line segment connecting the midpoints of the line segment M and the line segment N and the midpoint of the line segment K The point where the line extending in the axial direction intersects is defined as A, and a distance corresponding to 1/10 times the line segment connecting the respective midpoints of the line segment M and the line segment N from the midpoint of the line segment M. When the point moved in the developing direction is B, the value (A/B) obtained by dividing the anticoagulant adsorption amount at the point A by the anticoagulant adsorption amount at the point B is 0.5 to 1.0. , Blood purification column. Blood volume of the container is 50～220Cm ^3, packing density of the fibrous adsorption carrier is 0.2-0.5 g / cm ^3, a blood purification column according to claim 1, wherein. The blood purification column according to claim 1 or 2, wherein the concentration of the anticoagulant is 2 to 40 IU/cm ³ . The blood purification column according to claim 1, wherein the fibrous adsorption carrier is a knitted fabric or a woven fabric. The blood purification column according to any one of claims 1 to 4, wherein the anticoagulant is heparin, nafamostat mesylate, or low molecular weight heparin. The fiber constituting the fibrous adsorption carrier is a sea-island composite fiber,
The sea component is selected from the group consisting of polystyrene and its derivatives, polysulfone and its derivatives, and mixtures thereof;
The blood purification column according to any one of claims 1 to 5, wherein the island component is selected from the group consisting of polypropylene, polyethylene, polypropylene/polyethylene copolymer, and a mixture thereof.