Body fluid processing column, and use and production method of the same

Abstract

To provide a body fluid processing column which is excellent in adsorption performance of a hydrophobic substance, and suppresses column clogging even when being once exposed to air and air exists in the column, has high absorption performance and also can sterilize by high pressure steam, and a use and a production method of the same.SOLUTION: There is provided a body fluid processing column in which hydrophobic porous beads and filling liquid are filled. The body fluid processing column is configured so that air release property of the porous beads is 0.1 wt.% ethanol or more 11.0 wt.% ethanol or less, and a specific surface area of the porous beads in a dry state is 404 m2 / g or more and 970 m2 / g or less. The body fluid processing column further includes 0.3 vol.% to 6.0 vol.% of air in an internal part of the column, with an apparent volume of the porous beads as reference.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present disclosure relates to columns for treating body fluids, their uses and methods of manufacture. [Background technology]

[0002] Technology for treating biological fluids, such as blood, outside the body is being developed. The treated biological fluid can be used for various purposes, such as materials for producing diagnostic agents, therapeutic agents, vaccines, etc.; samples for analysis and research; and returning the biological fluid to the body for treatment (extracorporeal circulation). A biological fluid treatment column filled with porous beads and a filling liquid is often used for treating biological fluids. By passing the biological fluid through a biological fluid treatment column, desired substances can be adsorbed and removed from the biological fluid, and the treated biological fluid can be used for various purposes such as those mentioned above.

[0003] For example, in the treatment of ischemic diseases such as sepsis, various apheresis therapies are performed to remove inflammatory mediators, such as cytokines and alarmins, which are thought to be causative substances, from the patient's blood. In recent years, as one type of apheresis therapy, the development of adsorptive blood treatment columns, which remove inflammatory mediators by adsorption, has progressed.

[0004] Mechanical hemolysis often occurs during extracorporeal circulation in artificial heart-lung machines, artificial kidneys, and artificial livers, as well as during surgical operations. Mechanical hemolysis often causes hemoglobin to be liberated into plasma, which can lead to complications such as renal failure. The most common method for removing free hemoglobin is to administer fresh frozen plasma or a large amount of a haptoglobin preparation. In recent years, there has been progress in the development of adsorption-type blood treatment columns that remove free hemoglobin by adsorption during extracorporeal circulation.

[0005] Examples of adsorptive blood processing columns on the market include Toraymyxin (registered trademark) (Toray Medical Co., Ltd.), which uses an adsorbent consisting of a roll of fibers with endotoxin removal function; Sepxiris (registered trademark) (Baxter Japan Co., Ltd.), which is an adsorptive blood processing column for continuous blood purification therapy (CRRT) that uses hollow fibers with alarmin (HMGB1) and cytokine (IL-6, etc.) adsorption function; and CytoSorb (registered trademark) (Cytosorbents Corporation), which uses porous polymer beads with cytokine removal function.

[0006] Since blood processing columns are in direct contact with the patient's blood, they must be biocompatible. To impart biocompatibility to blood processing columns, the adsorbent is subjected to a bead surface treatment and coated with a biocompatible polymer, typically a hydrophilic polymer.

[0007] For example, Patent Document 1 aims to provide beads for blood treatment having improved blood compatibility while maintaining the adsorptivity of the porous beads, and describes beads for blood treatment having porous beads and a polymer supported on the surface of the porous beads. The polymer contains a monomer represented by a specific general formula as a monomer unit (Patent Document 1, Abstract, Claim 1, etc.).

[0008] Patent Document 2 aims to provide beads for blood treatment that are highly biocompatible and have little elution of the biocompatible polymer carried thereon into blood, and describes porous beads and beads for blood treatment having a polymer carried on the surface of the porous beads. The polymer contains an amphoteric monomer as a monomer unit, and the amphoteric monomer is 10 mol % or more and 30 mol % or less based on the total monomers constituting the polymer (Patent Document 2, Abstract, Claim 1, etc.).

[0009] Patent Document 3 aims to provide a system for reducing the level of a targeted compound in the blood, and describes an intravenous catheter with an in-line housing and a material within the housing that removes the targeted compound from the blood by selective adsorption. The material within the housing may include polymeric particles, which may include a coating that provides biocompatibility (Patent Document 3, Abstract, Claim 1, and paragraph 0029, etc.).

[0010] Patent document 4 aims to disclose a sorbent for hemoperfusion suitable for long-term clinical treatment and describes a polymer system comprising at least one polymer with multiple pores, said polymer being made hemocompatible by an external coating (Patent document 4, Abstract, and claims 1 and 15, etc.).

[0011] Patent document 5 aims to provide a size-selective, hemocompatible porous polymer adsorbent system that functions in direct contact with body fluids, in particular blood, and describes a polymer system comprising at least one polymer with a plurality of pores, the polymer being made hemocompatible by an external coating (Patent document 5, abstract, and claims 1 and 15, etc.).

[0012] Patent Document 6 describes a method for treating blood, blood products, plasma, or physiological fluids with the objectives of extending the useful life of blood products and improving the quality of blood, providing a biocompatible and hemocompatible system with improved performance in removing toxins, etc. The method includes contacting the blood, blood product, or physiological fluid with a sorbent, the sorbent being primarily in a solid form and including a crosslinked polymeric material having a plurality of at least one of (1) zwitterionic moieties and (2) oligo(ethylene glycol) moieties attached to a surface of the crosslinked polymeric material (Patent Document 6, Abstract, and Claim 1, etc.).

[0013] Patent Document 7 has as one of its objectives the provision of a porous body having improved blood compatibility while maintaining the adsorption properties of the porous body, and describes a blood purifier in which the low-melting point water content and contact change rate per gram of dry weight of the porous body are within specific ranges, and the longitudinal length L of the area occupied by the porous body in the main container and the circular equivalent diameter D of the cross section in the lateral direction satisfy a specific relationship.

[0014] Patent Document 8 aims to provide a blood purifier having a porous molded body that is excellent in blood compatibility, has good cytokine adsorption performance, has low pressure loss before and after blood processing, and is safe to use, and describes a blood purifier in which the low-melting point water content and contact change rate per gram of dry weight of the porous molded body are within specific ranges, and the amount of platelet adhesion per mL of blood when blood is brought into contact with the porous molded body is within a specific range.

[0015] Such adsorption-type blood processing columns are expected to be used in the treatment of ischemic diseases as well as in situations where excessive production of inflammatory mediators is a problem, such as cardiac surgery and organ transplant surgery. [Prior art documents] [Patent documents]

[0016] [Patent Document 1] JP 2020-006155 A [Patent Document 2] JP 2020-006156 A [Patent Document 3] JP 2009-078165 A [Patent Document 4] Special Publication No. 2013-537923 [Patent Document 5] JP 2017-125199 A [Patent Document 6] Special Publication No. 2016-514568 [Patent Document 7] International Publication No. 2020 / 203923 [Patent Document 8] International Publication No. 2020 / 203927 Summary of the Invention [Problem to be solved by the invention]

[0017] In general, hydrophobic porous beads have excellent adsorption performance for hydrophobic substances, and therefore can efficiently remove hydrophobic substances, such as inflammatory substances, from body fluids. However, a problem unique to hydrophobic and porous beads is their high adsorption performance for gases such as oxygen and nitrogen. When the beads have high gas adsorption performance, air tends to easily adhere to the beads when they come into contact with air. When air adheres to the hydrophobic porous beads, the effective adsorption area of ​​the beads is blocked by the air, and the adsorption performance is reduced. When manufacturing a column filled with hydrophobic porous beads and a filling liquid, air is often mixed into the column in the process of packing the beads into the column (filling process) and in subsequent processes, such as the column washing process, before use, and air is likely to adhere to the beads. Hydrophilic porous beads have good compatibility with water, so air can easily escape by simply immersing them in water, but hydrophobic porous beads easily repel water, so air cannot escape by simply immersing them in water. When hydrophobic porous beads with air attached thereto come into contact with bodily fluids, the coagulation reaction of the bodily fluids is accelerated, making the column more susceptible to clogging. For example, if the bodily fluid is blood, the air-attached portions become sites of thrombus formation, which can easily cause column clogging or an increase in plasma hemoglobin concentration due to hemolysis.

[0018] Normally, a body fluid treatment column is primed immediately before use to remove impurities contained in the filling solution in the column. The general priming method is to connect the body fluid treatment column to a dedicated blood purification tube and then wash it with about 2 L of pure water, physiological saline, or heparin solution. However, when connecting the column to the blood purification tube, air may flow into the column and adhere to the hydrophobic porous beads. Therefore, even if there is no air attached to the hydrophobic porous beads at the time of column production, a small amount of air may flow into the column immediately before use, causing adverse effects such as an increase in column pressure and a decrease in the adsorption performance of the beads.

[0019] As a method for removing air from hydrophobic porous beads to which air has adhered, it is effective to immerse the beads in a filling solution containing an effective amount of alcohol of 1000 ppm or more (hereinafter referred to as "high-alcohol-containing filling solution"). However, when a body fluid is treated using hydrophobic porous beads once immersed in a high-alcohol-containing filling solution, alcohol may be eluted into the treated body fluid, and alcohol has a harmful effect on the living body, so that the applications of the body fluid treatment column are significantly limited. For example, hydrophobic porous beads immersed in a high-alcohol-containing filling solution cannot be used at least for extracorporeal circulation applications in which the treated body fluid is returned to the living body, such as apheresis therapy.

[0020] Columns for treating body fluids are required to be sterilized during production, and one of the sterilization methods is high-pressure steam sterilization. In the case of a column packed with hydrophobic porous beads and a filling liquid, if there is a small amount of air inside the column, the air dissolved in the filling liquid will adhere to the beads. If the column is sterilized under high-pressure steam in such a state, the air attached to the beads will expand due to temperature changes during sterilization, creating a pressure difference between the inside and outside of the column, which can cause deformation or damage to the column. Therefore, when sterilizing a column packed with hydrophobic porous beads with high-pressure steam, it is necessary to thoroughly remove the air inside the column.

[0021] For these reasons, most conventional hydrophobic porous beads for treating body fluids have been hydrophilized to such an extent that air can be removed by simply immersing them in water. However, due to the hydrophilization, such hydrophobic porous beads have insufficient adsorption performance for hydrophobic substances.

[0022] Hydrophobic porous beads having a hydrophilic polymer material on the outer surface as described in Patent Documents 1 to 8 have improved biocompatibility derived from the hydrophilic polymer material. However, when porous hydrophobic adsorption beads come into contact with air, air still tends to adhere to the beads. When such beads are packed in a column and brought into contact with body fluids, the column is likely to be clogged or hemolyzed, and the adsorption performance of the beads tends to decrease. In addition, when the column is sterilized by high-pressure steam while air is present in the column, the column may be deformed or broken. Therefore, when manufacturing such a column, a process of removing air from the column is required, which increases the manufacturing cost of the column.

[0023] Therefore, one object of the present disclosure is to provide a body fluid treatment column that has excellent adsorption performance for hydrophobic substances, is less likely to become clogged even if exposed to air or if air is present in the column, has high adsorption performance, and is capable of being sterilized by high-pressure steam, as well as methods for using and manufacturing the same. [Means for solving the problem]

[0024] The present inventors have found that by adjusting the specific surface area and air release properties of the porous hydrophobic adsorbent beads, the beads can maintain their adsorption performance even when they come into contact with air, and that column clogging is unlikely to occur. Furthermore, they have found that by adjusting the specific surface area and air release properties of the porous hydrophobic adsorbent beads, a body fluid treatment column can be obtained that is unlikely to deform even after high-pressure steam sterilization, even if air is present in the column.

[0025] Examples of embodiments of the present disclosure are listed in the following items [1] to

[17] . [1] A body fluid treatment column packed with hydrophobic porous beads and a packing liquid, The porous beads have an air release property of 0.1% by weight ethanol or more and 11.0% by weight ethanol or less, The specific surface area of ​​the above porous beads in a dry state is 404 m 2 / g or more 970m 2 / g or less, The body fluid treatment column further contains 0.3 vol % to 6.0 vol % of air, based on the apparent volume of the porous beads, inside the column. [2] 2. A body fluid treatment column according to item 1, wherein the proportion of elemental sulfur in the porous beads is 0.01% by weight or more and 3.0% by weight or less by elemental analysis. [3] 3. A body fluid treatment column according to item 1 or 2, wherein the proportion of oxygen elements in the porous beads is 0.9% by weight or more and 5.1% by weight or less by elemental analysis. [4] 3. A body fluid treatment column according to item 1 or 2, wherein the porous beads are mainly composed of a styrene-based polymer. [5] 3. A body fluid treatment column according to item 1 or 2, wherein the proportion of carbon elements in the porous beads is 84.3% by weight or more and 90.7% by weight or less by elemental analysis. [6] 3. A body fluid treatment column according to item 1 or 2, wherein the porous beads are high-pressure steam sterilized porous beads, and when immersed in water, 50% by weight or more of the porous beads sink, based on the dry weight of the porous beads. [7] 3. The body fluid treatment column according to item 1 or 2, wherein the amount of alcohol contained in the filling solution is 0 ppm or more and less than 1000 ppm based on the total weight of the filling solution. [8] 3. The body fluid treatment column according to item 1 or 2, wherein the air leakage property of the porous beads is 0.1% by weight or more of ethanol and 9.0% by weight or less of ethanol. [9] The porous beads have a cumulative pore volume of 1 nm to 200 nm in a dry state of 0.5 cm 3 / g or more 3.5cm 3 3. The body fluid treatment column according to item 1 or 2, wherein the molecular weight of the column is 100 / g or less.

[10] 3. Use of a body fluid treatment column according to item 1 or 2, comprising contacting at least a portion of the porous beads with air before use of the body fluid treatment column.

[11] 3. A method for producing a body fluid treatment column according to item 1 or 2, comprising the step of modifying hydrophobic porous beads by irradiating the hydrophobic porous beads with gamma rays while the beads are immersed in an aqueous solution in which a radical reactive compound having a sulfur element is dissolved.

[12] Item 12. The method according to item 11, wherein the radical reactive compound comprises sulfurous acid or a salt thereof.

[13] Item 13. The method according to item 12, wherein the sulfurous acid or a salt thereof comprises at least one selected from the group consisting of sodium pyrosulfite, sodium hydrogensulfite, and sodium sulfite.

[14] Item 12. The method according to item 11, wherein the radical reactive compound comprises a compound having a thiol group or a disulfide group.

[15] Item 15. The method according to item 14, wherein the compound having a thiol group or a disulfide group further has at least one group selected from the group consisting of a carboxy group, an amino group, and a hydroxy group.

[16] Item 16. The method according to item 15, wherein the compound having a thiol group or a disulfide group has a molecular weight of 100 or more and 1,000 or less.

[17] Item 17. The method according to item 16, wherein the compound having a thiol group or a disulfide group includes at least one selected from the group consisting of thiomalic acid, lipoic acid, 3-mercaptopropionic acid, 3-methoxybutyl 3-mercaptopropionate, glutathione, dithiothreitol, cysteine, and cysteine ​​derivatives. Effect of the Invention

[0026] According to the present disclosure, there is provided a body fluid treatment column which has excellent adsorption performance for hydrophobic substances, is less likely to become clogged even when exposed to air or when air is present in the column, can maintain high adsorption performance, and can be sterilized with high-pressure steam, as well as methods for using and manufacturing the same. [Brief description of the drawings]

[0027] [Figure 1] FIG. 1 is a schematic diagram for explaining a test method for evaluating the adsorption rate of beads. [Diagram 2] FIG. 2 is a schematic diagram for explaining the blood compatibility evaluation method. [Diagram 3] FIG. 3 is a schematic diagram for explaining a method for cleaning a column packed with porous beads. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] <Body fluid treatment column> The body fluid treatment column of the present disclosure is a body fluid treatment column packed with hydrophobic porous beads (also simply referred to as "porous beads" in the present disclosure) and a filling liquid. The porous beads have a specific surface area of ​​404 m2 in a dry state. 2 / g or more 970m 2 / g or less, and has an air release property of 0.1% by weight ethanol or more and 11.0% by weight ethanol or less. The body fluid treatment column has a specific surface area of ​​404 m2 in a dry state of the hydrophobic porous beads. 2 / g or more, and the porous beads have an air release property of 0.1% by weight or more of ethanol, so that the beads have excellent adsorption performance for hydrophobic substances such as hemoglobin and have high biocompatibility. In addition, the specific surface area of ​​the porous beads in a dry state is 970 m 2 / g or less, and the air release property of 11.0 wt% ethanol or less makes it difficult for air to adhere even when it comes into contact with air, which prevents column clogging and hemolysis. If hemolysis can be suppressed, the increase in the concentration of free hemoglobin in blood generated by hemolysis can also be suppressed, and the original hemoglobin adsorption performance of the hydrophobic porous beads can be exhibited. Furthermore, even if air is present in the column, the beads' adsorption performance and biocompatibility can be maintained, and deformation of the column can be suppressed even when high-pressure steam sterilization is performed.

[0029] The body fluid is not limited as long as it is a liquid derived from a living organism, and examples thereof include blood, plasma, serum, ascites, lymph, intra-articular fluid, tissue fluid, digestive fluid, sweat, tears, nasal mucus, urine, semen, vaginal fluid, amniotic fluid, milk, etc., as well as fractionated components obtained therefrom. The treated body fluid can be used for various purposes, such as materials for producing diagnostic agents, therapeutic agents, vaccines, etc., samples for analysis and research, etc., and returning the body to a living organism for treatment, etc. (extracorporeal circulation).

[0030] The body fluid treatment column of the present disclosure is preferably a blood treatment column for treating blood, more preferably an extracorporeal circulation column for an artificial heart-lung machine, an artificial kidney, an artificial liver, etc., and even more preferably a blood treatment column for various apheresis therapies. In extracorporeal circulation treatment, mechanical hemolysis often occurs. Mechanical hemolysis increases the free hemoglobin concentration in the blood, which may lead to complications such as renal failure. In apheresis therapy, in general, for the treatment of ischemic diseases such as sepsis, inflammatory mediators, such as cytokines and alarmins, which are thought to be causative substances, are removed from the patient's blood and returned to the patient's body. In these applications, the advantages of the body fluid treatment column of the present disclosure are more pronounced. That is, in the manufacturing process of the blood treatment column, air may be mixed into the column when filling the column with porous beads or washing the column. Even if air is not mixed in during the manufacturing process, air may be mixed in the column during a priming operation (replacing the filling solution in the column with water, physiological saline, etc. and washing) performed immediately before using the column. When air is mixed in the column, the air adheres to the porous beads, which promotes blood coagulation and causes thrombus formation and hemolysis. In addition, the air blocks the effective adsorption area of ​​the beads, which causes a decrease in adsorption performance. In contrast, the body fluid treatment column of the present disclosure is less likely to have air adhered even when exposed to air, and can be used while maintaining its adsorption performance and biocompatibility, so that the concentration of free hemoglobin in the blood does not increase, and it can be preferably used for blood treatment applications, such as blood treatment applications for apheresis therapy.

[0031] <column> A body fluid treatment column has a column (main container) having at least a body fluid inlet, an internal space, and a body fluid outlet, and the internal space of the column contains porous beads and a filling liquid. During body fluid treatment, generally, the body fluid before treatment is introduced into the internal space through the body fluid inlet and treated by contacting with the porous beads present in the internal space, and the treated body fluid can flow out through the body fluid outlet.

[0032] The shape of the column is not limited, but may be, for example, a tubular, typically cylindrical, column.

[0033] Materials constituting the column include, but are not limited to, thermoplastic resins such as polypropylene, polyethylene, polyester, polystyrene, polytetrafluoroethylene, polycarbonate, acrylonitrile butadiene styrene (ABS), copolymers of vinyl aromatic hydrocarbons and conjugated dienes, etc. Thermosetting resins such as polyurethane and epoxy may also be used for sealing.

[0034] The volume of air that may be present in the column is preferably 0.3% by volume to 6.0% by volume, more preferably 0.5% by volume to 5.0% by volume, even more preferably 1.0% by volume to 5.0% by volume, even more preferably 2.0% by volume to 5.0% by volume, and particularly preferably 3.0% by volume to 5.0% by volume, based on the apparent volume of the hydrophobic porous beads in the column. Here, the "apparent volume" of the hydrophobic porous beads in the column does not mean the actual volume of the hydrophobic porous beads, but means the volume (volume) of the region including the gaps between the particles of the hydrophobic porous beads. In other words, the actual volume of the hydrophobic porous beads is the apparent volume of the hydrophobic porous beads in the column multiplied by the packing rate of the hydrophobic porous beads. By making the volume of air present in the column 6.0% by volume or less, it is possible to suppress the coagulation reaction from being accelerated due to the influence of the air present in the column or the air attached to the beads, thereby suppressing the pressure rise and hemolysis of the column. In addition, it is possible to suppress the effective adsorption area of ​​the beads from being blocked by air, thereby suppressing the decrease in adsorption performance. Furthermore, when the column is subjected to high-pressure steam sterilization, the pressure difference between the inside and outside of the column due to the expansion of the air attached to the beads can be reduced, so that deformation and damage of the column can be suppressed. On the other hand, allowing the presence of 0.3% by volume or more of air means that there is no need to strictly manage the column manufacturing process to prevent air from being mixed in, or to use degassed water or the like as a filling liquid, and the manufacturing cost of the column can be reduced. Even when air is present inside the column, the body fluid treatment column of the present disclosure is less likely to clog and can maintain high adsorption performance and biocompatibility. During the manufacture of a body fluid treatment column, a small amount of air often gets into the column, so by allowing the presence of air within the above range, it is possible to suppress column clogging and improve the productivity of the body fluid treatment column while maintaining high adsorption performance and biocompatibility.

[0035] <Hydrophobic porous beads> The hydrophobic porous beads of the present disclosure (also referred to simply as "porous beads" in the present disclosure) have numerous pores, and can adsorb hydrophobic substances into the pores by hydrophobic bonds. The hydrophobic porous beads have high adsorption performance for hydrophobic substances such as hydrophobic protein molecules, for example, free hemoglobin in blood generated during hemolysis, and inflammatory mediators such as cytokines and alarmins, and are therefore useful as porous beads for blood processing columns for adsorbing and removing these.

[0036] The hydrophobic porous beads may be any hydrophobic polymer capable of forming a porous carrier. Examples of hydrophobic polymers include sulfone-based polymers, ethersulfone-based polymers, vinylidene fluoride-based polymers, vinylidene chloride-based polymers, arylethersulfone-based polymers, ethylene-based polymers, propylene-based polymers, and styrene-based polymers. Among the above polymers, styrene-based polymers such as styrene-divinylbenzene copolymers are preferred because they are highly hydrophobic and have excellent thermal stability, acid resistance, alkali resistance, and mechanical strength. The hydrophobic porous beads are preferably composed mainly of styrene-based polymers. In the present disclosure, the term "main component" refers to the component that occupies the highest weight percent of the resin constituting the hydrophobic porous beads. The degree of polymerization and molecular weight of the polymer are not particularly limited. In the present disclosure, the term "styrene-based polymer" refers to a polymer containing styrene and / or substituted styrene as the main component of the monomer, and a polymer that has undergone further reaction with a polymer containing styrene and / or substituted styrene. Examples of further reactions include chloromethylation, sulfonation, and a reaction of introducing an amino group following chloromethylation.

[0037] The air release property of the porous beads is 0.1% by weight ethanol or more and 11.0% by weight ethanol or less, preferably 0.1% by weight ethanol or more and 9.0% by weight ethanol or less, more preferably 0.3% by weight ethanol or more and 9.0% by weight ethanol or less, even more preferably 0.3% by weight ethanol or more and 7.0% by weight ethanol or less, and particularly preferably 0.5% by weight ethanol or more and 5.0% by weight ethanol or less. The air release property of the porous beads can be controlled by the conditions and degree of the bead modification treatment. Conventional adsorbent beads for treating body fluids are excessively hydrophilized to an air release property of 0.0% by weight (air is judged to be released only by water), and have poor adsorption properties for hydrophobic substances, or have an air release property exceeding 11.0% by weight, so that once air is mixed in, the adsorption performance and biocompatibility are reduced (Patent Documents 1 to 8, and Comparative Examples 1 to 5, 8, and 9 of the present disclosure). In contrast, the porous beads of the present disclosure have an air release rate of 0.1% by weight or more, and therefore have high biocompatibility and excellent adsorption performance for hydrophobic substances. In addition, when the porous beads have an air release rate of 11.0% by weight or less, air is unlikely to adhere even when the beads come into contact with air during the process of packing the beads into a column or the priming operation of the column immediately before use, and therefore the adsorption performance and biocompatibility can be maintained at a high level. In addition, even if a small amount of air is present in a column packed with beads, deformation and damage of the column are unlikely to occur when the column is sterilized by high-pressure steam. In a preferred embodiment, when the porous beads have an air release rate of 9.0% by weight or less, air is hardly or never adhered during the process of packing the beads into a column or the priming operation of the column immediately before use, and therefore the adsorption performance and biocompatibility can be maintained at a high level.

[0038] In the present disclosure, the air release property (wt % ethanol) is a parameter measured by carrying out the following steps (1) to (5) as a pretreatment, followed by steps (6) to (8), starting with distilled water (ethanol concentration a=0 wt %) and increasing the ethanol concentration a by 1 wt %, and the ethanol concentration a at which it is determined that the sample first sinks is taken as the air release property (wt % ethanol). In this case, if the sample does not sink in distilled water but sinks in 1 wt % ethanol, the ethanol concentration a at which it is determined that the sample first sinks is taken as the air release property in 0.1 wt % increments. (1) Place 0.5 ml of porous beads into a 5 ml Eppendorf tube. (2) Add 4 ml of distilled water to the 5 ml Eppendorf tube (1) above and mix by inverting three times. (3) The solution contained in 0.5 ml of the beads from (2) above is removed using a cell strainer (Mini Cell Strainer II, nylon mesh 40 μm, manufactured by Funakoshi Co., Ltd.), and the obtained beads are placed in a 5 ml Eppendorf tube. (4) Repeat steps (2) and (3) above five times. (5) 0.5 ml of the porous beads obtained in (4) above is vacuum-dried at 50° C. for 15 hours (absolute pressure 0.003 MPa or less). (6) The porous beads vacuum dried in (5) above are immersed in distilled water containing a% by weight of ethanol (a≧0) and mixed by inversion at 1 rpm for 72 hours using a rotator (ROTATOR RT-5 (manufactured by Taitec Co., Ltd.)). (7) Leave it at room temperature for 1 hour. (8) Determine whether 50% or more by weight of the porous beads sinks to the bottom, based on their dry weight.

[0039] The specific surface area of ​​the porous beads in a dry state is 404 m 2 / g or more 970m 2 / g or less, preferably 450m 2 / g or more 950m 2 / g or less, more preferably 500m 2 / g or more 890m 2 / g or less, and even more preferably 530m 2 / g or more 850m2 / g or less, particularly preferably 550m 2 / g or more 800m 2 The specific surface area of ​​the porous beads in a dry state can be controlled by the conditions and the degree of the bead modification treatment. 2 / g or more, the adsorption performance of hydrophobic substances is improved, and the specific surface area in a dry state is 970 m 2 / g or less, it becomes easier to control the air release property to 9.0 wt % ethanol or less, air is less likely to adhere even after exposure to air, and biocompatibility can be maintained at a higher level.

[0040] The porous beads are required to be sterilized from the viewpoint of reducing adverse effects on living bodies. Sterilization methods include, but are not limited to, sterilization by heating, ultraviolet rays, radiation (e.g., gamma rays, electron beams), and the like. Sterilization by heating includes moist heat sterilization by steam or boiling, and dry heat sterilization by high-temperature dry air. Moist heat sterilization by steam includes, for example, high-pressure steam (autoclave) sterilization. From the viewpoint of reducing the burden on the environment, the porous beads are preferably porous beads sterilized by high-pressure steam. From the viewpoint of reducing adverse effects on living bodies, when the porous beads sterilized by high-pressure steam are immersed in water, it is preferable that 50% by weight or more of the porous beads sink, more preferably 60% by weight or more, even more preferably 70% by weight or more, and even more preferably 80% by weight or more sink, based on the dry weight of the porous beads. When the porous beads are sterilized by high-pressure steam while air is present in the column, the amount of air adhering to the beads increases further due to the air in the column being more likely to move during high-pressure steam sterilization, and air blockage may occur. For example, when blood is treated using porous beads with a large amount of air attached thereto, the blood may coagulate when it comes into contact with the air, causing an increase in the pressure of the blood flow. Conventional porous beads with an air release rate of 12% by weight or more do not sink when immersed in water, and it is understood that air is attached to the beads. In contrast, the porous beads in the preferred embodiment of the present disclosure are judged to sink when immersed in water when the air release rate is approximately 9% by weight or less. If the porous beads have an air release rate adjusted to a level where they sink by 50% by weight or more when immersed in water, air can be easily removed from the internal space of the porous beads and column without using a high-alcohol content filling solution even after high-pressure steam sterilization, and therefore high adsorption performance and biocompatibility can be maintained. In fact, referring to the examples and comparative examples of the present application, it can be seen that when the air release rate is 12.0% by weight or more, if there is air in the column, the column is likely to become clogged when blood is passed through it. It can also be seen that when the air release rate of the beads is 12.0% by weight or more, the adsorption performance cannot be maintained and the air block ratio of the beads is high.

[0041] In this context, whether or not the high-pressure steam sterilized porous beads sink in water is determined by the following steps (1) to (3). (1) After high-pressure steam sterilization, remove 8 to 10 mL of porous beads from the column. (2) The removed porous beads are immediately immersed in 90 mL of distilled water before they dry. (3) After immersion at room temperature for 30 minutes, determine whether or not 50% by weight or more of the porous beads sink to the bottom, based on the dry weight of the porous beads.

[0042] The harmonic mean particle diameter of the porous beads is preferably 300 μm to 1000 μm, more preferably 400 μm to 800 μm, and even more preferably 420 μm to 700 μm. When the harmonic mean particle diameter is 300 μm or more, the pressure rise when a body fluid is passed through a column can be effectively suppressed, and when the harmonic mean particle diameter is 1000 μm or less, rapid adsorption performance can be exhibited.

[0043] The cumulative pore volume of the porous beads having a pore diameter of 1 nm to 200 nm in a dry state is preferably 0.5 cm 3 / g or more 3.5cm 3 The lower limit of the cumulative pore volume is more preferably 0.8 cm 3 / g or more, more preferably 1.0 cm 3 / g or more, and even more preferably 1.4 cm 3 The upper limit of the cumulative pore volume, which can be arbitrarily combined with these lower limits, is more preferably 3.0 cm 3 / g or less, more preferably 2.5 cm 3 / g or less, particularly preferably 2.0 cm 3 When the cumulative pore volume is within the above range, the adsorptivity of the porous beads is further improved, and the porous beads can remove a larger number of hydrophobic protein molecules, which is preferable.

[0044] In a preferred embodiment of the present disclosure, the hydrophobic porous beads preferably have an oxygen element ratio of 0.9% by weight or more and 5.1% by weight or less, more preferably 1.0% by weight or more and 5.0% by weight or less, even more preferably 1.0% by weight or more and 4.0% by weight or less, and particularly preferably 1.0% by weight or more and 3.2% by weight or less, as determined by elemental analysis. By having an oxygen element ratio of 5.1% by weight or less, the adsorption performance of the hydrophobic substance is improved. In addition, by having an oxygen element ratio of 0.9% by weight or more, it is easier to control the air release property to 11.0% by weight ethanol or less, and air is less likely to adhere even after contact with air, so that the adsorption performance and biocompatibility can be maintained at a higher level.

[0045] Of the elements constituting the entire porous beads, the proportion of carbon element is preferably 84.3% by weight or more and 90.7% by weight or less, and more preferably 85.6% by weight or more and 90.7% by weight or less, as determined by elemental analysis. By having a carbon element proportion of 84.3% by weight or more, the adsorption performance of hydrophobic substances is improved. By having a carbon element proportion of 90.7% by weight or less, it is easier to control the air release property to 11.0% by weight ethanol or less, air is less likely to adhere even after contact with air, and the adsorption performance and biocompatibility can be maintained at a high level.

[0046] Among the elements constituting the entire porous beads, the proportion of sulfur element is preferably 0.01% by mass or more and 3.0% by mass or less, more preferably 0.01% by mass or more and 2.5% by mass or less, even more preferably 0.01% by mass or more and 2.0% by mass or less, and even more preferably 0.2% by mass or more and 1.6% by mass or less, as determined by elemental analysis. By having the proportion of sulfur element of 3.0% by weight or less, the adsorption performance of hydrophobic substances is improved. By having the proportion of sulfur element of 0.01% by weight or more, it becomes easier to control the air release property to 11.0% by weight ethanol or less, and air is less likely to adhere even after contact with air, so that the adsorption performance and biocompatibility can be maintained at a higher level.

[0047] The proportion of nitrogen element among the elements constituting the entire porous beads is preferably more than 0% by mass and 0.8% by mass or less, more preferably more than 0% by mass and 0.4% by mass or less, as determined by elemental analysis. When the proportion of nitrogen element is within the above range, the porous beads tend to have high adsorption performance for hydrophobic substances and high biocompatibility, which is preferable.

[0048] <Adsorption properties of porous beads> The porous beads of the present disclosure preferably tend to have improved adsorption properties when removing hydrophobic substances, such as hydrophobic protein molecules, having a molecular weight of more than 500 Da and less than 70,000 Da from body fluids. In the present disclosure, "capable of removing" a certain hydrophobic substance means that when a body fluid sample containing the hydrophobic substance to be removed is brought into contact with the porous beads and shaken, the adsorption rate of the hydrophobic substance to the porous beads is 20% or more. More preferably, the porous beads are capable of removing hydrophobic protein molecules having a molecular weight of more than 1,000 Da and less than 70,000 Da. For example, free hemoglobin has a molecular weight of approximately 64.5 kDa, cytokines have molecular weights of approximately 5 to 60 kDa (IL-1b: approximately 17.5 kDa, IL-6: approximately 24.5 kDa, IL-8: approximately 8 kDa, IL-10 (dimer): approximately 37.5 kDa, TNF-α (trimer): approximately 51 kDa), and the alarmin high mobility group box 1 (HMGB1) is a hydrophobic protein with a molecular weight of approximately 30 kDa.

[0049] The hydrophobic protein molecules to be removed include protein molecules thought to be the cause of sepsis, such as exogenous substances derived from pathogenic microorganisms, such as PAMPs (pathogen-associated molecular patterns); and various inflammatory mediators leading to inflammatory reactions, such as alarmins, which are endogenous substances released due to tissue damage, and cytokines, which cause inflammatory reactions. Other hydrophobic protein molecules include free hemoglobin in the blood, which is released into the blood during hemolysis.

[0050] Examples of PAMPs include endotoxin (LPS), peptidoglycan (PGN), lipoteichoic acid, double-stranded RNA (dsRNA), and flagellin.

[0051] Examples of alarmins include high mobility group box 1 (HMGB1), heat shock proteins (HSPs), histones, fibrinogen, neutrophil elastase, and macrophage migration inhibitory factor (MIF).

[0052] Examples of cytokines include interleukins (IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, and IL-18) and tumor necrosis factors (TNF-α, TNF-β). Of these, the porous beads preferably remove alarmins and cytokines, and more preferably remove HMGB1 and cytokines.

[0053] The porous beads preferably have an adsorption rate of TNF-α of 41% or more and 100% or less, more preferably 45% to 100%, and even more preferably 50% to 100%, when a body fluid sample containing a hydrophobic substance to be removed is contacted with the porous beads having a volume 0.1 times that of the body fluid sample and shaken at 37° C. for 6 hours. A TNF-α adsorption rate of 41% or more is expected to provide a higher therapeutic effect as a body fluid treatment column.

[0054] The hemoglobin (Hb) adsorption rate of the porous beads is preferably 31% or more and 80% or less, more preferably 40% to 80%, even more preferably 44% to 80%, and even more preferably 49% to 80%. If the hemoglobin adsorption rate is within the above range, the free hemoglobin in blood generated by hemolysis can be efficiently adsorbed. In general, when air adheres to hydrophobic porous beads, the effective adsorption area of ​​the beads is blocked by the air, and the adsorption performance is reduced. The effect of air adsorption on the beads on the reduction in adsorption performance can be indirectly measured by measuring the hemoglobin adsorption rate of the beads after immersing them in 20% ethanol by once, and comparing the hemoglobin adsorption rates before and after immersion in 20% ethanol. When the value obtained by dividing the "hemoglobin adsorption rate before immersion in ethanol" by the "hemoglobin adsorption rate after immersion in ethanol" is 85% or more, it can be determined that the beads have little air blockage.

[0055] Biocompatibility of Porous Beads The porous beads of the present disclosure have excellent adsorptivity as described above, and also have high biocompatibility, and can maintain high biocompatibility even after exposure to air. The term "biocompatibility" varies depending on the purpose and use of the body fluid treatment column, but when the body fluid treatment column is a blood treatment column, it can be judged by the time it takes for the pressure in the column to increase. Generally, when blood is continuously flowed, blood coagulation gradually occurs in the column, the flow resistance of the column gradually increases, and as a result, the column pressure increases. However, the higher the biocompatibility, the less likely blood coagulation occurs, and therefore the less likely the pressure in the column to increase when blood is flowed through the column.

[0056] <Filling liquid> The amount of alcohol contained in the filling solution of the body fluid treatment column of the present disclosure is preferably 0 ppm or more and less than 1000 ppm, more preferably 0 ppm to 300 ppm, even more preferably 0 ppm to 100 ppm, still more preferably 0 ppm to 30 ppm, and particularly preferably 0 ppm to 10 ppm, based on the total weight of the filling solution. By having the amount of alcohol within the above range, adverse effects on the living body are further reduced. Furthermore, by having the amount of alcohol in the filling solution of 0 ppm or more and less than 1000 ppm, adverse effects on the living body are small, preferably almost or completely absent. Therefore, it can be safely used in applications where adverse effects on the living body are a concern, such as blood treatment applications.

[0057] As the base liquid for the filling liquid, liquids generally used in body fluid treatment columns can be used, and typical examples include ultrapure water and physiological saline.

[0058] <<Use of columns for treating body fluids>> The present disclosure also provides a use of the body fluid treatment column of the present disclosure, which includes contact of air with at least a portion of the porous beads before use. The use of the body fluid treatment column preferably includes, for example, a step of washing (priming) the body fluid treatment column before use. When air comes into contact with at least a portion of the porous beads, air may adhere to the porous beads, which may cause a decrease in biocompatibility and a decrease in adsorption performance. When blood is treated using porous beads with air attached thereto, the blood may coagulate when it comes into contact with the air, causing an increase in blood flow pressure. In such a use involving contact with air, the advantages of the body fluid treatment column of the present disclosure are more pronounced. That is, the porous beads of the present disclosure are less likely to have air attached thereto even when exposed to air, and therefore can maintain high biocompatibility even in applications involving contact of at least a portion of the porous beads with air.

[0059] The body fluid treatment column is more preferably used for blood treatment including contacting at least a portion of the porous beads with air before use, and even more preferably used for blood treatment for various apheresis therapies, because the body fluid treatment column of the present disclosure has high biocompatibility even after the porous beads come into contact with air, and the alcohol content in the filling solution is low, resulting in less adverse effects on the living body.

[0060] <<Method for manufacturing a body fluid treatment column>> The method for producing a body fluid treatment column of the present disclosure includes a step of subjecting hydrophobic porous beads to a bead modification treatment, and a step of packing the porous beads and a packing liquid into a column. In a preferred embodiment, the method for producing a body fluid treatment column includes a step of subjecting hydrophobic porous beads to a bead modification treatment, a step of packing the porous beads and a packing liquid into a column, and a step of sterilizing the porous beads, in this order. Sterilization is preferably high-pressure steam sterilization.

[0061] <Beads modification process> The method of bead modification is to increase the specific surface area of ​​the porous beads to 404 m 2 / g or more 970m 2 / g or less, and the air escape property can be controlled to 0.1 wt% ethanol or more and 11.0 wt% ethanol or less. As the bead modification treatment, for example, at least a part of the hydrophobic porous beads may be bonded with another material. Preferably, at least a part of the hydrophobic porous beads is physically or chemically modified. More preferably, at least one element selected from the group consisting of sulfur element, oxygen element, and nitrogen element is introduced into the hydrophobic porous beads, and even more preferably, sulfur element is introduced into the hydrophobic porous beads. In terms of ease of adjusting the air escape property of the porous beads within the range of the present disclosure, it is preferable to modify not only the outer surface of the hydrophobic porous beads but also the inner surface of the pores.

[0062] As a method for modifying hydrophobic porous beads, for example, the specific surface area and air release properties of the hydrophobic porous beads can be adjusted by impregnating the hydrophobic porous beads with a filling solution containing a compound (radical reactive compound) having a sulfur element and a substituent that generates radicals when irradiated with gamma rays, and irradiating the beads with gamma rays. Examples of compounds that generate radicals when irradiated with gamma rays include azo compounds, organic peroxides, and sulfurous acid or its salts. Examples of substituents that generate radicals when irradiated with gamma rays include hydroxyl groups, epoxy groups, amino groups, thiol groups, disulfide groups, vinyl groups, benzyl groups, allyl groups, halogenated alkyl groups, enols, and tertiary carbon atoms.

[0063] Among them, sulfurous acid or its salt, thiol group, and disulfide group are more preferred in that they are highly reactive under gamma-ray irradiation and therefore the specific surface area and air release property of the hydrophobic porous beads can be easily adjusted. Examples of sulfurous acid or its salt include at least one selected from the group consisting of sodium pyrosulfite, sodium hydrogensulfite, and sodium sulfite. The molecular weight of the compound having a thiol group or disulfide group is preferably 100 or more and 1000 or less in terms of increasing the reactivity under gamma-ray irradiation. The compound having a thiol group or disulfide group is preferably further provided with at least one group selected from the group consisting of a carboxy group, an amino group, and a hydroxy group as its functional group in that it has better blood compatibility.

[0064] From the viewpoints of reactivity, blood compatibility, and availability, compounds having a substituent that generates radicals upon gamma-ray irradiation are preferably sulfite, thiomalic acid, lipoic acid, 3-mercaptopropionic acid, 3-methoxybutyl 3-mercaptopropionate, glutathione, dithiothreitol, cysteine, and cysteine ​​derivatives. Examples of cysteine ​​derivatives include acetylcysteine, methyl cysteine, ethyl cysteine, propyl cysteine, and cystine, as well as low molecular weight peptides having cysteine ​​as one of the amino acids.

[0065] From the viewpoint of facilitating the introduction of sulfur element by gamma ray irradiation in the presence of a radically reactive compound having sulfur element, and therefore facilitating the adjustment of the specific surface area and air release property to the ranges of the present disclosure, it is preferable that the hydrophobic porous beads are mainly composed of a styrene-based polymer. The details of the styrene-based polymer are as described above.

[0066] <Sterilization process> The manufacturing method of a body fluid treatment column is required to include a sterilization step in order to reduce adverse effects on the living body. Sterilization methods include, but are not limited to, sterilization by heating, ultraviolet rays, radiation (e.g., gamma rays, electron beams), and the like. Sterilization by heating includes moist heat sterilization by steam or boiling, and dry heat sterilization by high-temperature dry air. Moist heat sterilization by steam includes, for example, high-pressure steam sterilization. From the viewpoint of reducing the burden on the environment, the porous beads are preferably sterilized by high-pressure steam sterilization. Since the porous beads of the present disclosure are unlikely to have air attached to them even when they come into contact with air, even if high-pressure steam sterilization is performed in a state in which air is present in the column, the column is unlikely to deform or break, and high adsorption performance and biocompatibility can be maintained.

[0067] <Column packing process> The column may be any of those described in the column above. A body fluid treatment column may be produced by packing a column with hydrophobic porous beads, the specific surface area and air release properties of which have been adjusted, and a packing liquid. The porous beads described in the column above may be used as the packing liquid. EXAMPLES

[0068] <<Measurement of the physical properties of porous beads>> <Air release properties of porous beads> The air release property (wt% ethanol) is a parameter measured by carrying out the following steps (1) to (5) as a pretreatment, followed by steps (6) to (8), in which the ethanol concentration a is increased by 1 wt% starting from distilled water (ethanol concentration a = 0 wt%), and the ethanol concentration a at which it is determined that the sample first sinks is taken as the air release property (wt% ethanol). In the case where the sample does not sink in distilled water but sinks in 1 wt% ethanol, the ethanol concentration a at which it is determined that the sample first sinks is taken as the air release property in 0.1 wt% increments. (1) Place 0.5 ml of porous beads into a 5 ml Eppendorf tube. (2) Add 4 ml of distilled water to the 5 ml Eppendorf tube (1) above and mix by inverting three times. (3) The solution contained in 0.5 ml of the beads from (2) above is removed using a cell strainer (Mini Cell Strainer II, nylon mesh 40 μm, manufactured by Funakoshi Co., Ltd.), and the obtained beads are placed in a 5 ml Eppendorf tube. (4) Repeat steps (2) and (3) above five times. (5) 0.5 ml of the porous beads obtained in (4) above is vacuum-dried at 50° C. for 15 hours (absolute pressure 0.003 MPa or less). (6) The porous beads vacuum dried in (5) above are immersed in distilled water containing a% by weight of ethanol (a≧0) and mixed by inversion at 1 rpm for 72 hours using a rotator (ROTATOR RT-5 (manufactured by Taitec Co., Ltd.)). (7) Leave it at room temperature for 1 hour. (8) Determine whether 50% or more by weight of the porous beads sinks to the bottom, based on their dry weight.

[0069] <Harmonic mean particle size of porous beads> The size of the porous beads swollen with ultrapure water was measured by image analysis using CAMSIZER X2 (Microtrackbell), and their harmonic mean particle diameter (μm) was calculated.

[0070] <Specific surface area and cumulative pore volume of porous beads> The porous beads swollen with ultrapure water were frozen and then freeze-dried for 24 hours. After drying the porous beads, they were degassed (dried under reduced pressure) at 60°C for 15 hours using a VacPrep061 (Shimadzu Micromeritics). Thereafter, the specific surface area (m2) was measured by nitrogen gas adsorption using a TriStarII 3020 (Shimadzu Micromeritics). 2 / g) and cumulative pore volume (cm 3 In this case, the BET plot value was used as the specific surface area, and the desorption cumulative pore volume according to the BJH method was used as the cumulative pore volume.

[0071] Elemental analysis of porous beads The solution contained in 1 mL of porous beads was removed using a cell strainer, and the resulting beads were added to a 15 mL tube. Then, 14 mL of ultrapure water was added to the 15 mL tube. This series of steps was repeated five times to replace the bead solution with ultrapure water. The porous beads replaced with ultrapure water were vacuum dried at 50°C for 15 hours (absolute pressure 0.003 MPa or less). Elemental analysis of oxygen, nitrogen, and carbon was performed on the dried porous beads using an elemental analyzer (HORIBA, Ltd., oxygen, nitrogen, and hydrogen analyzer EMGA-930). Elemental analysis of sulfur was performed on the dried porous beads using an ion chromatograph (Thermo Fisher Scientific, INTEGRION) by the oxygen flask combustion method. Three samples were analyzed for each test, and the average value was used.

[0072] <TNF-α adsorption rate of porous beads> Figure 1 is a schematic diagram for explaining the bead adsorption evaluation test method. After adding heparin sodium (heparin sodium injection 50,000 units / 50mL, manufactured by Nipro) to blood collected from a healthy volunteer to a concentration of 2000 IU / L, lipopolysaccharide (LPS) derived from Escherichia coli O111:B4 (manufactured by Sigma-Aldrich) was added to a concentration of 0.1 μg / mL, and the mixture was shaken at 37°C for 24 hours at a shaking angle of 10 degrees and 10 r / min using a shaker (Invitro Shaker WAVE-S1, manufactured by TAITEC). After that, the mixture was centrifuged at 2000 g for 20 minutes at room temperature using a centrifuge (Hybrid High-Speed ​​Refrigerated Centrifuge 6200, manufactured by Kubota Shoji Co., Ltd.), and the supernatant was obtained as a plasma sample. 9.0 mL of the obtained plasma sample and 0.90 mL of porous beads (approximately 0.20 g when dried) were mixed in a 15 mL tube made of polypropylene (PP). As shown in FIG. 1, the tube (32) was attached radially along the radial direction of the rotator on a 20 cm diameter disk-shaped rotor (31) of a rotator (30) (ROTATOR RT-5 (manufactured by Taitec Co., Ltd.)). The angle of the rotating surface of the disk-shaped rotor was set to 90 degrees from the horizontal, and the mixture was rotated and stirred at a speed of 4 rpm for 6 hours at 37°C (this is called the bead-contacted sample). At this time, a sample was also prepared in which beads were not added to the obtained 9.0 mL plasma sample, and the same treatment as for the bead-contacted sample was performed (this is called the non-bead-contacted sample). The PP tube after rotation and stirring was centrifuged at room temperature at 2000 g for 1 minute using a centrifuge, and the supernatants of the bead-contacted and non-bead-contacted samples were obtained. The TNF-α concentration of the obtained supernatant was measured using a Bio-Plex system (Bio-Rad Bio-Plex Pro human cytokine GI8-plex A panel) according to the attached instruction manual. The TNF-α adsorption rate was calculated using the following formula. TNF-α adsorption rate (%) = ("TNF-α concentration in sample without contact with beads" - "TNF-α concentration in sample with contact with beads") / "TNF-α concentration in sample without contact with beads" x 100 In this experiment, the TNF-α concentration without contact with beads was 1906 pg / mL.

[0073] <Hemoglobin (Hb) adsorption rate of porous beads> A hemoglobin solution was prepared by adding 300 mg of hemoglobin (bovine, Fujifilm Wako Pure Chemical Industries, Ltd.) and 35 g of albumin (derived from bovine serum, fatty acid-free, Fujifilm Wako Pure Chemical Industries, Ltd.) to 1.0 L of 1XPBS(-) (composition: 137 mmol / L NaCl, 8.1 mmol / L Na2HPO4, 2.68 mmol / L KCl, 1.47 mmol / L KH2PO4, pH 7.4, Fujifilm Wako Pure Chemical Industries, Ltd.) and completely dissolving them. 9.0 mL of the obtained hemoglobin solution and 0.90 mL of porous beads (about 0.20 g when dried) were mixed in a 15 mL tube made of polypropylene (PP). As shown in FIG. 1, the tubes (32) were attached radially along the radial direction of the rotating body on the disk-shaped rotating body (31) of a rotator (30) (ROTATOR RT-5, Taitec Co., Ltd.) with a diameter of 20 cm. The angle of the rotating surface of the disk-shaped rotor was set to 90 degrees from the horizontal, and the whole was shielded from light and rotated at 4 rpm for 2 hours at 22°C (this is the sample with contact with beads). At this time, a sample was also prepared in which no beads were added to the obtained hemoglobin 9.0 mL, and the same treatment as the sample with contact with beads was performed (this is the sample without contact with beads). The PP tube after rotation and stirring was centrifuged at 2000 g at room temperature for 1 minute using a centrifuge to obtain the supernatant of the sample with and without contact with beads. The obtained supernatant was diluted 3.00 times with 1XPBS(-), and the absorbance (406 nm) of the 3-fold diluted solution against 1XPBS(-) was measured using a Shimadzu UV-2600 ultraviolet-visible spectrophotometer, and the hemoglobin adsorption rate was calculated using the following formula. Hemoglobin adsorption rate (%) = ("absorbance of 3-fold diluted solution of sample without contact with beads" - "absorbance of 3-fold diluted solution of sample with contact with beads") / "absorbance of 3-fold diluted solution of sample without contact with beads" x 100 In addition, the absorbance of the 3-fold diluted solution of the sample not in contact with the beads in this experiment was 0.662.

[0074] <Air blocking ratio of porous beads> Using a 15 mL tube, 1 mL of beads was immersed in 10 mL of 20 wt% ethanol (prepared from distilled water for injection (Otsuka distilled water, manufactured by Otsuka Pharmaceutical Factory) and special reagent grade ethanol (manufactured by Fujifilm Wako Pure Chemical Industries)) and left to stand for 1 hour. Then, 9.5 mL of the solution was removed, 9.5 mL of distilled water for injection was added, and the solution was left to stand for 24 hours. The 9.5 mL of solution was removed again, 9.5 mL of distilled water for injection was added, and the solution was left to stand for 24 hours to prepare beads. The hemoglobin adsorption rate of the beads after the preparation was measured to determine the hemoglobin adsorption rate of the beads after immersion in 20 wt% ethanol. When the value obtained by dividing the "hemoglobin adsorption rate of the beads before immersion in 20 wt% ethanol" by the "hemoglobin adsorption rate after immersion in 20 wt% ethanol" was 85% or more, it was determined that the beads had little air blockage, and when it was less than 85%, it was determined that the beads had a lot of air blockage.

[0075] <Alcohol concentration of column packing solution> The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the liquid packed in the porous beads-packed column were measured by the following procedures (1) to (3). (1) The liquid packed in a column packed with porous beads is removed by forcing air through the column. (2) The obtained filling solution is diluted 10 times with distilled water for injection (Otsuka distilled water, manufactured by Otsuka Pharmaceutical Factory), and the components of the diluted solution are detected with a flame ionization detector (FID) using a gas chromatograph (GC-2014, manufactured by Shimadzu Corporation, packing material: SUMPAK-H). (3) Compare with the peak area detected for methanol, ethanol, n-propanol, isopropanol, and ethylene glycol, each adjusted to 100 ppm with distilled water for injection.

[0076] <Water immersion test of steam-sterilized porous beads> Whether or not the high-pressure steam sterilized porous beads sink in water was determined by the following steps (1) to (3). (1) After high-pressure steam sterilization, remove 8 to 10 mL of porous beads from the column. (2) The removed porous beads are immediately immersed in 90 mL of distilled water before they dry. (3) After immersion for 30 minutes at room temperature, judge whether or not 50% or more by weight of the porous beads sink to the bottom based on their dry weight. If 50% or more by weight sinks, it is rated as "good," and if 50% or more by weight does not sink, it is rated as "poor."

[0077] <Blood compatibility test of porous beads> FIG. 2 is a schematic diagram for explaining the blood compatibility evaluation method. 5.0 mL (about 1.1 g when dried) of porous beads taken out from the column after high-pressure steam sterilization was immersed in physiological saline (Otsuka saline injection, manufactured by Otsuka Pharmaceutical Factory). The swollen porous beads (11) were packed into a 5.0 mL syringe (made of PP, inner diameter 13φ). At this time, as shown in FIG. 2, the top and bottom of the porous beads were sandwiched with a mesh (12) (opening 183 um, aperture ratio 42%) and a silicon O-ring (13) to prevent the beads from leaking, and the inside of the column was filled with physiological saline. Finally, 50 μL of air was mixed in from the top of the column using a pipette, creating a column with 1.0 vol. % air. In this way, a mini-column (10) packed with 5.0 mL of porous beads (11) was created.

[0078] Heparin sodium (heparin sodium injection 50,000 units / 50 mL, manufactured by Nipro) was added to blood collected from healthy pigs weighing 50 kg or more and 70 kg or less to a concentration of 1500 IU / L (this is referred to as "untreated blood"). Next, an experimental circuit was assembled as shown in FIG. 2, and physiological saline (Otsuka saline injection, manufactured by Otsuka Pharmaceutical Factory) was passed through a mini-column (10) packed with 5.0 mL of porous beads (11) at a flow rate of 6.0 mL / min for 10 minutes using a peristaltic pump (21) (SJ-1211II, manufactured by ATTO) so that the liquid flowed from the top of the column to the bottom of the column. At this time, the untreated blood (21) was placed in a thermostatic bath (23) and kept at a temperature of 37°C. It was also confirmed that the value of the manometer (22) (PG-200-102GP, manufactured by Nidec Copal Electronics) was -1.0 kPa or more and 2.0 kPa or less. Next, while gently stirring the untreated blood with a stirrer (24) to make it homogenous, the blood was passed through the column at a temperature of 37°C and a flow rate of 6.0 mL / min using a peristaltic pump (21) (SJ-1211II, manufactured by ATTO) so that the liquid flowed from the top to the bottom of the column. If the manometer pressure exceeded 40 kPa within 40 minutes after the blood was passed through, the blood was stopped at that point, and the blood compatibility was determined to be poor. If the manometer pressure did not exceed 40 kPa within 40 minutes after the blood was passed through, the blood was stopped at 40 minutes, and the blood compatibility was determined to be good.

[0079] <Air volume in the column> The amount of air in the column can be measured by the following steps (1) to (13). (1) Hold the side of the column with your hand so that the blood inlet is facing upwards, and gently tap the bottom of the column with your hand to push the air that has accumulated at the bottom of the column to the top of the column. (2) Clamp the column while keeping the blood inlet facing upwards. (3) Place a vibrator (MD-011-YA, manufactured by THRIVE) against the side of the column, set the vibrator to strong, and switch it on. (4) The vibrator is moved up and down for 1 minute to vibrate the column and collect air near the blood inlet of the column. (5) Remove the vibrator from the column and switch the vibrator off. (6) While keeping the blood inlet of the column facing upwards, rotate the column 90 degrees counterclockwise when viewed from above. (7) Repeat the series of steps (3) to (6) a total of four times to collect the air inside the column at the blood inlet of the column. (8) Prepare a PVC tube with a connector that can be connected to a column, and connect the PVC tube to a 50 mL syringe. (9) Fill a 50 mL syringe connected to a PVC tube with 30 mL of saline (Otsuka Saline Injection, manufactured by Otsuka Pharmaceutical Factory) and ensure that no air remains inside the 50 mL syringe or the PVC tube. (10) Open the plug on the blood inlet side of the column and connect the column to the PVC tubing, taking care not to let any more air in. (11) Raise the 50 mL syringe above the height of the column and slowly pull the inner cylinder of the syringe to create negative pressure inside the column, thereby moving the air inside the column into the syringe. (12) After removing your hands from the inner tube of the syringe and waiting for a while, slowly pull back on the inner tube of the syringe again repeatedly to move all of the air in the column into the syringe. (13) After removing your hand from the inner cylinder of the syringe and allowing it to stand for a while, read the amount of air remaining in the syringe and use this amount as the amount of air in the column.

[0080] Example 1 <Bead modification treatment of hydrophobic porous beads> Muromac was used as the porous beads before the bead modification treatment. TM SAP-9210 (Muromachi Chemical Co., Ltd., styrene-based polymer beads, harmonic mean particle diameter 440 μm) was used.

[0081] 200 mL of porous beads (apparent volume 200 mL, approximately 44.4 g when dry) before the bead modification treatment was placed in a 1.0 L polypropylene (PP) container (eye boy pp wide-mouth bottle 1 L), and then 800 mL of 57% by weight ethanol (prepared from distilled water for injection (Otsuka distilled water, Otsuka Pharmaceutical Factory) and special grade ethanol (Fujifilm Wako Pure Chemical)) was added. After shaking for 12 hours at a shaking angle of 10 degrees and 40 r / min using a shaker (Invitro Shaker WAVE-S1, TAITEC), the solution after shaking was filtered using a polyester mesh (TB50, mesh size 308 um, Fluorochemical Co., Ltd.). The absorbance of the filtered solution at 220 nm was measured using a Shimadzu ultraviolet-visible spectrophotometer UV-2600 (Shimadzu Corporation), and the beads obtained by filtration were added again to the 1.0 L PP container. This series of steps, which consisted of adding 57% by weight ethanol to the 1.0 L container, shaking for 12 hours on a shaker, and removing the solution through a polyester mesh, was repeated until the absorbance at 220 nm of the filtered solution was between -0.03 and 0.03 relative to the absorbance of the 57% by weight ethanol used.

[0082] Next, 200 mL of the beads after washing with 57 wt% ethanol were placed in a 1.0 L polypropylene (PP) container, and then 800 mL of distilled water for injection was added. After shaking for 1 hour at a shaking angle of 10 degrees and 40 r / min using a shaker, the solution after shaking was filtered using a polyester mesh (TB50). The absorbance of the filtered solution at 220 nm was measured using a Shimadzu UV-2600 ultraviolet-visible spectrophotometer, and the beads obtained by filtration were added again to the 1.0 L PP container. The series of operations of adding distilled water for injection to this 1.0 L container, shaking with a shaker for 1 hour, and removing the solution using a polyester mesh were repeated until the absorbance of the filtered solution at 220 nm was between -0.03 and 0.03 relative to the absorbance of distilled water for injection.

[0083] Of the 200 mL of beads obtained, 150 mL was placed in a 500 mL PP container (Eye-Boy PP wide-mouth bottle 500 mL), after which 350 mL of a 0.18 wt % aqueous solution of sodium pyrosulfite prepared from distilled water for injection and sodium pyrosulfite (Tokyo Chemical Industry Co., Ltd.) was added, and the beads were modified by γ-ray irradiation (30.8 kGy).

[0084] 150 mL of beads after the bead modification treatment were filtered using a polyester mesh (TB50) to remove the solution. The filtered beads were placed in a 1.0 L PP container, and 850 mL of saline (Otsuka saline injection, manufactured by Otsuka Pharmaceutical Factory) was added. After shaking for 1 hour at a shaking angle of 10 degrees and 40 r / min using a shaker, the shaken beads were filtered using a polyester mesh (TB50). The beads obtained by filtration were added again to the 1.0 L PP container. The series of steps of adding saline to this 1.0 L container, shaking for 1 hour using a shaker, and removing the solution using a polyester mesh was repeated a total of 7 times.

[0085] <Physical properties and performance evaluation of porous beads> The physical properties and performance evaluation of the above porous beads was carried out by the method described in the above section "Measurement of Physical Properties of Porous Beads". The air escape property was 7.0% by weight ethanol (in the table, simply indicated as "%". The same applies below). The results of elemental analysis were 90.0% carbon, 8.1% hydrogen, less than 0.3% nitrogen, 1.1% oxygen, and 0.20% sulfur. The specific surface area of ​​the beads was 650 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.83 cm 3 The TNF-α adsorption rate of these beads was 51%.

[0086] <Preparation of a column packed with porous beads sterilized by high-pressure steam> FIG. 3 is a schematic diagram for explaining a method for cleaning a porous bead-packed column. 50 mL of the 150 mL of beads after bead modification and cleaning was used with physiological saline (Otsuka Saline Injection, manufactured by Otsuka Pharmaceutical Factory) to create a cylindrical column (10) made of polypropylene (PP) (inner diameter 35φ, volume 50 mL, polyester mesh (12) (opening 183 um, aperture ratio 42%) on the upper and lower surfaces), porous beads (11), and physiological saline. Next, an experimental circuit was assembled as shown in FIG. 3, and the physiological saline was kept at a temperature of 37° C. in a thermostatic bath (23) and gently stirred with a stirrer (24) to make it uniform, and passed through the porous bead-packed column (10) containing air at a flow rate of 20.0 mL / min for 15 minutes using a peristaltic pump (21) (SJ-1211II, manufactured by ATTO). Next, 0.5 mL of air was mixed into the porous bead-packed column (10) by pushing out the packed liquid using a pipette (Eppendorf, 1 mL), and the column was capped and gently inverted to mix, after which two porous bead-packed columns containing 1.0 vol% air were prepared in total. The porous bead-packed columns containing air were sterilized at 121°C for 60 minutes in a high-pressure steam sterilizer (LSX-500, TOMY) and left to stand in an environment of 20 to 25°C for 3 weeks.

[0087] <Performance evaluation of columns packed with porous beads sterilized by high-pressure steam> No deformation or damage was observed in the column after high-pressure steam sterilization. The performance of the column and beads after sterilization was evaluated using the method described in the above section "Measurement of the physical properties of porous beads." The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the filling liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 57%. The air blocking rate of the beads was determined to be low. The results of the water immersion test of the beads were good (over 80%), and the blood compatibility was also good (40 min).

[0088] Example 2 The same operations as in Example 1 were carried out, except that in the <Beads modification treatment of hydrophobic porous beads>, the concentration of sodium pyrosulfite added before gamma-ray irradiation was 0.30% by weight, and in the <Preparation of a column packed with porous beads sterilized by high-pressure steam>, 1.0 mL of air was mixed in to make a column packed with porous beads containing 2.0% by volume of air. When the <Evaluation of the physical properties and performance of the porous beads> was carried out, the air escape property was 5.0% by weight of ethanol. The results of elemental analysis were 89.3% carbon, 8.1% hydrogen, less than 0.3% nitrogen, 1.3% oxygen, and 0.35% sulfur. The specific surface area of ​​the beads was 643 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.81 cm 3 / g. The TNF-α adsorption rate of these beads was 54%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 61%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and their blood compatibility was also good (40 min).

[0089] Example 3 The same operations as in Example 1 were carried out, except that in the <bead modification treatment of hydrophobic porous beads> the concentration of sodium pyrosulfite added before gamma-ray irradiation was 1.20% by weight, and in the <preparation of a column packed with porous beads sterilized by high-pressure steam> 1.5 mL of air was mixed in to make a column packed with porous beads containing 3.0% by volume of air. When the <evaluation of the physical properties and performance of the porous beads> was carried out, the air escape property was 0.3% by weight of ethanol. The results of elemental analysis were 88.3% carbon, 8.0% hydrogen, less than 0.3% nitrogen, 2.0% oxygen, and 0.53% sulfur. The specific surface area of ​​the beads was 631 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.74 cm 3 / g. The TNF-α adsorption rate of these beads was 60%. When a performance evaluation of a column packed with porous beads sterilized by high-pressure steam was performed, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 65%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and blood compatibility was also good (40 min).

[0090] Example 4 The same operations as in Example 1 were carried out, except that in the <Beads modification treatment of hydrophobic porous beads>, the concentration of sodium pyrosulfite added before gamma-ray irradiation was 0.09% by weight. When the <Evaluation of physical properties and performance of porous beads> was carried out, the air escape property was 10.0% by weight of ethanol. The results of elemental analysis were carbon 90.7%, hydrogen 8.1%, nitrogen less than 0.3%, oxygen 1.0%, and sulfur 0.09%. The specific surface area of ​​the beads was 664 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.91 cm 3 / g. The TNF-α adsorption rate of these beads was 45%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 52%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and their blood compatibility was also good (40 min).

[0091] Example 5 The same operations as in Example 1 were carried out, except that in the <Beads modification treatment of hydrophobic porous beads>, the solution added before gamma-ray irradiation was not a 0.18 wt% aqueous solution of sodium pyrosulfite, but a 200 mM aqueous solution of thiomalic acid prepared from distilled water for injection and thiomalic acid (manufactured by Tokyo Chemical Industry Co., Ltd.), and in the <Preparation of a column packed with porous beads sterilized by high-pressure steam>, 2.5 mL of air was mixed to prepare a column packed with porous beads containing 5.0 vol% air. When the <Evaluation of the physical properties and performance of the porous beads> was carried out, the air escape property was 0.4 wt% ethanol. The results of the elemental analysis were carbon 85.6%, hydrogen 7.7%, nitrogen less than 0.3%, oxygen 4.0%, and sulfur 1.6%. The specific surface area of ​​the beads was 529 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.38 cm 3 / g. The TNF-α adsorption rate of these beads was 50%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 64%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and their blood compatibility was also good (40 min).

[0092] Example 6 The same operations as in Example 1 were carried out, except that in the <Beads modification treatment of hydrophobic porous beads>, the solution added before gamma-ray irradiation was not a 0.18 wt% aqueous solution of sodium pyrosulfite, but a 25 mM aqueous solution of acetylcysteine ​​prepared from distilled water for injection and N-acetyl-L-cysteine ​​(manufactured by Tokyo Chemical Industry Co., Ltd.). When the <Evaluation of physical properties and performance of porous beads> was carried out, the air escape property was 8.0 wt% ethanol. The results of elemental analysis were carbon 89.9%, hydrogen 8.1%, nitrogen 0.3%, oxygen 1.2%, and sulfur 0.40%. The specific surface area of ​​the beads was 601 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.77 cm3 / g. The TNF-α adsorption rate of these beads was 51%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 52%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and their blood compatibility was also good (40 min).

[0093] Example 7 The same operations as in Example 1 were carried out, except that in the <Beads modification treatment of hydrophobic porous beads>, the solution added before gamma-ray irradiation was not a 0.18 wt% aqueous solution of sodium pyrosulfite, but a 200 mM aqueous solution of acetylcysteine ​​prepared from distilled water for injection and N-acetyl-L-cysteine ​​(manufactured by Tokyo Chemical Industry Co., Ltd.), and in the <Preparation of a column packed with porous beads sterilized by high-pressure steam>, 2.5 mL of air was mixed to prepare a column packed with porous beads containing 5.0 vol% air. When the <Evaluation of the physical properties and performance of the porous beads> was carried out, the air escape property was 0.1 wt% ethanol. The results of the elemental analysis were 87.2% carbon, 7.9% hydrogen, 0.4% nitrogen, 2.6% oxygen, and 1.2% sulfur. The specific surface area of ​​the beads was 561 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.52 cm 3 / g. The TNF-α adsorption rate of these beads was 56%. When a performance evaluation of a column packed with porous beads sterilized by high-pressure steam was performed, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 60%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and blood compatibility was also good (40 min).

[0094] Example 8 The same operations as in Example 1 were carried out except that in the <Beads modification treatment of hydrophobic porous beads>, the solution added before γ-ray irradiation was not a 0.18 wt% aqueous solution of sodium pyrosulfite, but a 50 mM aqueous solution of D,L-α lipoic acid prepared from distilled water for injection and D,L-α lipoic acid (manufactured by Tokyo Chemical Industry Co., Ltd.), and in the <Preparation of a column packed with porous beads sterilized by high-pressure steam>, 1.0 mL of air was mixed to make a column packed with porous beads containing 2.0% by volume of air. When the <Evaluation of the physical properties and performance of the porous beads> was carried out, the air release rate was 6.0 wt% ethanol. The results of the elemental analysis were 88.9% carbon, 8.0% hydrogen, less than 0.3% nitrogen, 2.0% oxygen, and 1.1% sulfur. The specific surface area of ​​the beads was 575 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.62 cm 3 / g. The TNF-α adsorption rate of these beads was 57%. When a performance evaluation of a column packed with porous beads sterilized by high-pressure steam was performed, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 55%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and blood compatibility was also good (40 min).

[0095] Example 9 The same operations as in Example 1 were carried out, except that in the <Beads modification treatment of hydrophobic porous beads>, the solution added before gamma-ray irradiation was not a 0.18 wt% aqueous solution of sodium pyrosulfite, but a 200 mM aqueous solution of glutaraldehyde prepared from distilled water for injection and glutaraldehyde (manufactured by Tokyo Chemical Industry Co., Ltd.). When the <Evaluation of the physical properties and performance of the porous beads> was carried out, the air escape property was 9.0 wt% ethanol. The results of elemental analysis were carbon 90.4%, hydrogen 8.1%, nitrogen less than 0.3%, oxygen 1.2%, and sulfur less than 0.01%. The specific surface area of ​​the beads was 605 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.72 cm 3 / g. The TNF-α adsorption rate of these beads was 55%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 49%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and their blood compatibility was also good (40 min).

[0096] Example 10 In the "Beads modification treatment of hydrophobic porous beads", diamond ions are used as the porous beads before the beads modification treatment. TM The same operations as in Example 1 were carried out, except that HP20 (Mitsubishi Chemical Corporation, styrene-based polymer beads, harmonic mean particle size 420 μm) was used and the concentration of sodium pyrosulfite added before gamma-ray irradiation was 0.60% by weight. When the physical properties and performance evaluation of the porous beads was carried out, the air escape property was 3.0% by weight of ethanol. The results of elemental analysis were carbon 89.8%, hydrogen 8.0%, nitrogen less than 0.3%, oxygen 1.1%, and sulfur 0.32%. The specific surface area of ​​the beads was 618 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.06 cm 3 / g. The TNF-α adsorption rate of these beads was 55%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 61%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and their blood compatibility was also good (40 min).

[0097] Example 11 In the "Beads modification treatment of hydrophobic porous beads", Amberlite was used as the porous beads before the beads modification treatment. TM The same operations as in Example 1 were carried out, except that FPX66 (Organo Corporation, styrene-based polymer beads, harmonic mean particle size 740 μm) was used, the solution added before gamma-ray irradiation was not a 0.18 wt% aqueous solution of sodium pyrosulfite but a 130 mM aqueous solution of reduced glutathione prepared from distilled water for injection and reduced glutathione (Tokyo Chemical Industry Co., Ltd.), and 1.5 mL of air was mixed in the <Preparation of a column packed with porous beads sterilized by high-pressure steam> to prepare a column packed with porous beads containing 3.0% by volume of air. When the <Evaluation of the physical properties and performance of the porous beads> was carried out, the air release rate was 2.0 wt% ethanol. The results of elemental analysis were 88.6% carbon, 7.9% hydrogen, 0.3% nitrogen, 2.1% oxygen, and 0.57% sulfur. The specific surface area of ​​the beads was 894 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.70 cm 3 / g. The TNF-α adsorption rate of these beads was 48%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 53%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and their blood compatibility was also good (40 min).

[0098] Comparative Example 1 Synthesis of Coating Polymer A copolymer of 2-methoxyethyl methacrylate (MEMA, compound of formula (i)), N,N-diethylaminoethyl methacrylate (DEAEMA, compound of formula (ii)), and N-methacryloyloxyethyl-N,N-dimethylammonium-α-N-methylcarboxybetaine (CMB, compound of formula (iii)) was synthesized by normal solution polymerization. The polymerization conditions were as follows: in an ethanol solution, in the presence of 0.0025 mol / L of azoisobutyronitrile (AIBN) as an initiator, the concentration of each monomer was 1 mol / L, and the polymerization reaction was carried out at a reaction temperature of 60°C for 8 hours to obtain a polymer polymerization liquid. The obtained polymer polymerization liquid was dropped into diethyl ether, and the precipitated polymer was collected. The collected polymer was purified by performing a reprecipitation operation using diethyl ether. The obtained polymer was then dried under reduced pressure for 24 hours to obtain a coating polymer.

[0099] The molar ratio of the MEMA monomer unit, the DEAEMA monomer unit, and the CMB monomer unit in the coating polymer was measured as follows: The obtained coating polymer was dissolved in dimethyl sulfoxide, and then, 1 The calculation was made from the area ratio of the peak at 4.32 ppm (attributed to H atoms specific to CMB) and the peak at 2.63 ppm (attributed to H atoms specific to DEAEMA) in the chart calculated by H-NMR measurement, and the peak at 0.65-2.15 ppm (total H atom amount) using the following formula. Molar ratio of DEAEMA monomer = ("area ratio of 2.63 ppm region" / 2) / ("area ratio of 0.65-2.15 ppm region" / 5-"area ratio of 2.63 ppm region" x 0.3) x 100 Molar ratio of CMB monomer = ("area ratio of 4.32 ppm region" / 2) / ("area ratio of 0.65-2.15 ppm region" / 5-"area ratio of 2.63 ppm region" x 0.3) x 100 Molar ratio of MEMA monomer = 100 - Molar ratio of DEAEMA monomer - Molar ratio of CMB monomer The molar ratio of MEMA, DEAEMA, and CMB monomer units in the coating polymer was calculated to be 80 / 10 / 10.

[0100] Preparation of Coating Solution The above coating polymer was added to 57% by weight of ethanol, and then stirred for 12 hours to prepare a coating liquid with a coating polymer concentration of 0.2% by weight.

[0101] <Bead modification treatment of hydrophobic porous beads> Amberlite was used as porous beads before the bead modification treatment. TM XAD TM 1180N (Organo Corporation, styrene-based polymer beads, harmonic mean particle size 480 μm) was used. 200 mL of porous beads (apparent volume 200 mL, approximately 44.4 g when dried) before bead modification treatment was placed in a 1.0 L polypropylene (PP) container (eye-boy pp wide-mouth bottle 1 L), and then 800 mL of 57 wt% ethanol (prepared from distilled water for injection (Otsuka distilled water, Otsuka Pharmaceutical Factory) and special reagent grade ethanol (Fujifilm Wako Pure Chemical Industries)) was added. After shaking for 12 hours at a shaking angle of 10 degrees and 40 r / min using a shaker (Invitro Shaker WAVE-S1, TAITEC Corporation), the solution after shaking was filtered using a polyester mesh (TB50, mesh size 308 um, Fluorochemical Corporation). The absorbance of the filtered solution at 220 nm was measured using a Shimadzu UV-2600 ultraviolet-visible spectrophotometer (Shimadzu Corporation), and the beads obtained by filtration were added back to a 1.0 L PP container. The series of steps of adding 57% by weight ethanol to the 1.0 L container, shaking for 12 hours using a shaker, and removing the solution using a polyester mesh were repeated until the absorbance of the filtered solution at 220 nm was between -0.03 and 0.03 relative to the absorbance of the 57% by weight ethanol used.

[0102] 150 mL of the 200 mL beads obtained by the above treatment was placed in a 1 L PP container, and then 600 mL of the above coating solution was added, and the container was shaken for 3 hours at a shaking angle of 10 degrees and 40 r / min using a shaker. The solution was then filtered using a polyester mesh (TB50). The 1 L container containing the coated beads was then vacuum dried at 50°C for 15 hours (absolute pressure 0.003 MPa or less), and 600 mL of 20% by weight ethanol was added to the 1 L container. The container was shaken for 12 hours at a shaking angle of 10 degrees and 40 r / min using a shaker, and the solution after shaking was filtered using a polyester mesh (TB50).

[0103] Next, 150 mL of the beads obtained by the above treatment were placed in a 1 L PP container, and 750 mL of distilled water for injection was added. After shaking for 1 hour at a shaking angle of 10 degrees and 40 r / min using a shaker, the solution after shaking was filtered using a polyester mesh (TB50). The absorbance of the filtered solution at 220 nm was measured using a Shimadzu UV-2600 ultraviolet-visible spectrophotometer, and the beads obtained by filtration were added again to a 1 L PP container. A series of operations including adding distilled water for injection to this 1 L container, shaking for 1 hour using a shaker, and removing the solution using a polyester mesh were repeated until the absorbance of the filtered solution at 220 nm was -0.03 or more and 0.03 or less relative to the absorbance of the distilled water for injection. The obtained beads were placed in a 500 mL PP container, and 350 mL of distilled water for injection was added, followed by gamma irradiation (28.6 kGy).

[0104] The beads after gamma ray treatment were filtered using a polyester mesh (TB50) to remove the solution. The filtered beads were placed in a 1.0 L PP container, and 850 mL of saline (Otsuka saline injection, manufactured by Otsuka Pharmaceutical Factory) was added. The beads were shaken for 1 hour at a shaking angle of 10 degrees and 40 r / min using a shaker, and then filtered using a polyester mesh (TB50). The beads obtained by filtration were added again to the 1.0 L PP container. The series of steps of adding saline to this 1.0 L container, shaking using a shaker for 1 hour, and removing the solution using a polyester mesh was repeated a total of 7 times.

[0105] The beads were used to carry out the evaluation of the properties and performance of porous beads described in Example 1. The air release rate was 12.0% by weight of ethanol. Elemental analysis showed that the carbon content was 90.8%, the hydrogen content was 8.2%, the nitrogen content was less than 0.3%, the oxygen content was 0.9%, and the sulfur content was less than 0.01%. The specific surface area of ​​the beads was 612 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.59 cm 3 / g. The TNF-α adsorption rate of these beads was 34%. A column was created as described in <Creation of a column packed with high-pressure steam sterilized porous beads>, and a performance evaluation of a column packed with high-pressure steam sterilized porous beads> was performed, revealing no deformation or damage to the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 38%. The air block rate of the beads was determined to be high. The results of the water immersion test of the beads were poor (less than 50%), and the blood compatibility was slightly poor (38 min).

[0106] Comparative Example 2 The same operations as in Comparative Example 1 were carried out except that in <Synthesis of coating polymer>, the composition of the coating polymer was BMA (n-butyl methacrylate, compound of structural formula (iv)) / CMB=70 / 30 (molar ratio), and in <Preparation of high-pressure steam sterilized porous bead packed column>, 1.5 mL of air was mixed to make a porous bead packed column containing 3.0 volume % air. When <Evaluation of the physical properties and performance of the porous beads> was carried out, the air escape property was 12.0 wt % ethanol. The measurement results of elemental analysis were carbon 90.8%, hydrogen 8.2%, nitrogen less than 0.3%, oxygen 0.9%, and sulfur less than 0.01%. The specific surface area of ​​the beads was 595 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.58 cm 3 / g. The TNF-α adsorption rate of these beads was 33%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 37%. The air blocking rate of the beads was determined to be high. The results of the water immersion test of the beads were poor (less than 50%), and their blood compatibility was somewhat poor (36 min).

[0107] Comparative Example 3 The same operations as in Comparative Example 1 were carried out, except that in <Preparation of coating solution>, PVP (Polyvinylpyrrolidone K90, Fujifilm Wako Pure Chemical Industries, Ltd.) was used as the coating polymer, and in <Preparation of high-pressure steam sterilized porous bead-packed column>, 0.25 mL of air was mixed to make a porous bead-packed column containing 0.5% by volume of air. When <Evaluation of the physical properties and performance of the porous beads> was carried out, the air escape property was 12.0% by weight of ethanol. The results of elemental analysis were carbon 90.9%, hydrogen 8.2%, nitrogen less than 0.3%, oxygen 0.8%, and sulfur less than 0.01%. The specific surface area of ​​the beads was 622 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.62 cm 3 / g. The TNF-α adsorption rate of these beads was 37%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 36%. The air block rate of the beads was determined to be high. The results of the water immersion test of the beads were poor (less than 50%), and the blood compatibility was also poor (29 min).

[0108] Comparative Example 4 The same operations as in Example 1 were carried out, except that in the <bead modification treatment of hydrophobic porous beads>, the solution added before gamma-ray irradiation was distilled water for injection, instead of 0.18 wt% aqueous sodium pyrosulfite solution. When the <physical properties and performance evaluation of porous beads> was carried out, the air escape property was 12.0 wt% ethanol. The results of elemental analysis were carbon 91.0%, hydrogen 8.1%, nitrogen less than 0.3%, oxygen 0.7%, and sulfur less than 0.01%. The specific surface area of ​​the beads was 655 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.88 cm 3 / g. The TNF-α adsorption rate of these beads was 40%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 44%. The air block rate of the beads was determined to be high. The results of the water immersion test of the beads were poor (less than 50%), and the blood compatibility was also poor (28 min).

[0109] Comparative Example 5 In <Synthesis of coating polymer>, the composition of the coating polymer is MEMA=100. In <Preparation of coating solution>, the coating polymer concentration is 0.5% by weight. In <Bead modification treatment of hydrophobic porous beads>, Muromac is used as the porous beads before the bead modification treatment. TM The same procedure as in Comparative Example 1 was carried out, except that SAP-9210 was used and the solution added before gamma-ray irradiation was a 300 mM aqueous thiomalic acid solution prepared from distilled water for injection and thiomalic acid. When the physical properties and performance evaluation of the porous beads was carried out, the air escape property was 0.0 wt% ethanol. The results of elemental analysis were carbon 84.3%, hydrogen 7.8%, nitrogen less than 0.3%, oxygen 5.1%, and sulfur 1.6%. The specific surface area of ​​the beads was 403 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 0.79 cm 3 / g. The TNF-α adsorption rate of these beads was 25%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 30%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%), and their blood compatibility was also good (40 min).

[0110] Comparative Example 6 The same operations as in Example 4 were carried out, except that in the <Preparation of a column packed with high-pressure steam sterilized porous beads>, 3.0 mL of air was mixed in to make the column packed with porous beads containing 6.1% by volume of air. <Evaluation of the physical properties and performance of the porous beads> was as described in Example 4. When the <Evaluation of the performance of the column packed with high-pressure steam sterilized porous beads> was carried out, expansion (deformation) was observed in the header part of the column after high-pressure steam sterilization. The hemoglobin adsorption rate of the beads was 44%. The air block rate of the beads was determined to be high.

[0111] Comparative Example 7 The same procedure as in Example 11 was carried out, except that in the <bead modification treatment of hydrophobic porous beads>, the solution added before gamma-ray irradiation was a 0.09 wt% aqueous solution of sodium pyrosulfite, rather than a 130 mM aqueous solution of reduced glutathione. <Evaluation of the physical properties and performance of the porous beads> was carried out, and the air escape property was 10.0 wt% ethanol. The results of elemental analysis were carbon 90.6%, hydrogen 8.1%, nitrogen less than 0.3%, oxygen 0.9%, and sulfur 0.05%. The specific surface area of ​​the beads was 971 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 1.98 cm 3 / g. The TNF-α adsorption rate of these beads was 33%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 46%. The air blocking rate of the beads was determined to be high. The results of the water immersion test of the beads were poor (less than 50%), and the blood compatibility was also poor (26 min).

[0112] Comparative Example 8 In the "Beads modification treatment of hydrophobic porous beads", diamond ions are used as the porous beads before the beads modification treatment. TM The same operations as in Example 1 were carried out, except that HPA25L (Mitsubishi Chemical Corporation, styrene-based polymer beads, harmonic mean particle size 460 μm) was used, and the solution added before gamma-ray irradiation was distilled water for injection, not a 0.60 wt % aqueous solution of sodium pyrosulfite. When <Evaluation of the physical properties and performance of the porous beads> was carried out, the air escape property was 0.0 wt % ethanol. The results of elemental analysis were carbon 72.4%, hydrogen 8.2%, nitrogen 4.2%, oxygen 0.8%, and sulfur less than 0.01%. The specific surface area of ​​the beads was 31 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 0.14 cm 3 / g. The TNF-α adsorption rate of these beads was 11%. When a performance evaluation was performed on a column packed with porous beads that had been sterilized by high-pressure steam, no deformation or damage was found in the column. The concentrations of methanol, ethanol, n-propanol, isopropanol, and ethylene glycol in the packed liquid were all below 1000 ppm. The hemoglobin adsorption rate of the beads was 7%. The air blocking rate of the beads was determined to be low. The results of a water immersion test of the beads were good (over 80%).

[0113] Comparative Example 9 In the "Beads modification treatment of hydrophobic porous beads", diamond ions are used as the porous beads before the beads modification treatment. TMThe same operations as in Example 1 were carried out, except that PK228L (Mitsubishi Chemical Corporation, styrene-based polymer beads, harmonic mean particle size 720 μm) was used, and the solution added before gamma-ray irradiation was distilled water for injection, not a 0.60 wt % aqueous solution of sodium pyrosulfite. An evaluation of the physical properties and performance of the porous beads was carried out, and the air escape rate was 0.0 wt % ethanol. The results of elemental analysis were carbon 47.1%, hydrogen 4.0%, nitrogen less than 0.3%, oxygen 21.3%, and sulfur 10.0%. The specific surface area of ​​the beads was 1 m 2 / g, and the cumulative pore volume for pores with diameters of 1 nm to 200 nm is 0.01 cm 3 The TNF-α adsorption rate of these beads was 20%.

[0114] [Table 1] [Industrial Applicability]

[0115] The body fluid treatment column of the present disclosure can be used as a column for treating body fluids for various applications, such as materials for producing diagnostic agents, therapeutic agents, vaccines, etc.; samples for analysis and research, etc.; and returning body fluids to a living body (extracorporeal circulation) for treatment, etc. The body fluid treatment column of the present disclosure can be preferably used as a blood treatment column for treating blood, and more preferably as a blood treatment column for extracorporeal circulation, for example, various apheresis therapies. [Explanation of symbols]

[0116] 10 Columns 11 Porous beads 12 Mesh 13 O-ring 21 Peristaltic Pump 22 Manometer 23 Constant temperature bath 24 Stirrer 30 Rotator 31 Rotating disc 32 Tube

Claims

1. A body fluid treatment column packed with hydrophobic porous beads and a packing liquid, The porous beads have an air release property of 0.1% by weight ethanol or more and 11.0% by weight ethanol or less; The specific surface area of ​​the porous beads in a dry state is 404 m ２ / g or more 970m ２ / g or less, The body fluid treatment column further contains 0.3 vol % to 6.0 vol % of air based on the apparent volume of the porous beads inside the column.

2. 2. The body fluid treatment column according to claim 1, wherein the proportion of elemental sulfur in the porous beads is 0.01% by weight or more and 3.0% by weight or less by elemental analysis.

3. 3. The body fluid treatment column according to claim 1, wherein the proportion of oxygen elements in the porous beads is 0.9% by weight or more and 5.1% by weight or less, as determined by elemental analysis.

4. 3. The body fluid treatment column according to claim 1, wherein the porous beads are mainly composed of a styrene-based polymer.

5. 3. The body fluid treatment column according to claim 1, wherein the proportion of carbon elements in the porous beads is 84.3% by weight or more and 90.7% by weight or less, as determined by elemental analysis.

6. 3. The body fluid treatment column according to claim 1, wherein the porous beads are high-pressure steam sterilized porous beads, and when immersed in water, 50% by weight or more of the porous beads sink, based on the dry weight of the porous beads.

7. 3. The body fluid treatment column according to claim 1, wherein the amount of alcohol contained in the filling liquid is 0 ppm or more and less than 1000 ppm based on the total weight of the filling liquid.

8. 3. The body fluid treatment column according to claim 1, wherein the air leakage property of the porous beads is 0.1% by weight or more and 9.0% by weight or less of ethanol.

9. The porous beads have a cumulative pore volume of pores with diameters of 1 nm to 200 nm in a dry state of 0.5 cm ３ / g or more 3.5cm ３ The body fluid treatment column according to claim 1 or 2, wherein the molecular weight of the column is 1 / g or less.

10. 3. Use of a body fluid treatment column according to claim 1 or 2, comprising contacting at least a portion of the porous beads with air before use of the body fluid treatment column.

11. 3. A method for manufacturing a body fluid treatment column according to claim 1 or 2, comprising the step of modifying hydrophobic porous beads by irradiating the hydrophobic porous beads with gamma rays while the hydrophobic porous beads are immersed in an aqueous solution in which a radical reactive compound having a sulfur element is dissolved.

12. The method of claim 11 , wherein the radical reactive compound comprises sulfurous acid or a salt thereof.

13. 13. The method of claim 12, wherein the sulfurous acid or salt thereof comprises at least one selected from the group consisting of sodium pyrosulfite, sodium bisulfite, and sodium sulfite.

14. The method of claim 11 , wherein the radical reactive compound comprises a compound having a thiol group or a disulfide group.

15. The method according to claim 14, wherein the compound having a thiol group or a disulfide group further has at least one group selected from the group consisting of a carboxy group, an amino group, and a hydroxy group.

16. The method according to claim 15, wherein the compound having a thiol group or a disulfide group has a molecular weight of 100 or more and 1,000 or less.

17. The method according to claim 16, wherein the compound having a thiol group or a disulfide group includes at least one selected from the group consisting of thiomalic acid, lipoic acid, 3-mercaptopropionic acid, 3-methoxybutyl 3-mercaptopropionate, glutathione, dithiothreitol, cysteine, and cysteine ​​derivatives.

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