Converting time modulators of blood specimens deficient in coagulation factor VIII, IX or XI and reagents for determining activated partial thromboplastin time

Abstract

The present invention provides a blood coagulation time regulator for a blood sample deficient in blood coagulation factor VIII, IX or XI in activated partial thromboplastin time measurement, the blood coagulation time regulator containing, as an active ingredient, a polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit. Provided is a reagent for measuring activated partial thromboplastin time, which contains a polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit and an amino acid.

Description

Technical Field

[0001] The present invention relates to a coagulation time regulator for a blood sample lacking coagulation factor VIII, IX or XI in an activated partial thromboplastin time (APTT) assay, a reagent for APTT assay, and an APTT assay method using the same. Background Art

[0002] Coagulation test is a test used to diagnose the patient's coagulation ability by adding a specified reagent to the patient's blood sample and measuring the coagulation time. Typical examples of coagulation time include prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time, etc. Abnormal coagulation ability causes the extension of coagulation time. The causes of the extension of coagulation time include coagulation inhibitors (heparin, etc.), a decrease in components involved in coagulation, congenital coagulation factor deficiency (hemophilia, etc.), autoantibodies that inhibit coagulation reactions (lupus anticoagulant: LA, anticoagulation factor antibodies, etc.), etc.

[0003] In a coagulation test, a reagent is added to a blood sample, the subsequent coagulation reaction is measured, and the coagulation time is determined by the coagulation reaction. When a prolonged coagulation time is confirmed, it is determined to be an abnormal coagulation ability. Coagulation test reagents contain components for causing coagulation under certain conditions. For example, APTT assay reagents contain coagulation factor activators such as ellagic acid, phospholipids, buffers, etc. On the other hand, there are various types of commercially available APTT assay reagents, and the sensitivity to various coagulation abnormalities is different among these reagents.

[0004] Patent document 1 discloses an APTT assay reagent containing manganese chloride as an active enhancer and glycine as a suspension enhancement means. Patent document 2 records that aluminum chloride, ferric chloride, or a mixture of manganese chloride and ellagic acid activates the coagulation of the sample, but ferric chloride generates a precipitate of ellagic acid. Patent document 3 records the use of glycylglycine or diglycylglycine as a coagulation factor stabilizer in the APTT assay, but if they are added at a high concentration, the coagulation time is prolonged. Patent document 4 records that Good's buffers such as HEPES, PIPES, POPSO, ACES, TAPSO, EPPS, Tricine, and CHES can be used as specimen diluents for coagulation ability assays. Patent document 5 records that an APTT assay reagent containing specified concentrations of phosphatidylcholine, phosphatidylserine, and phosphatidylethanolamine has improved sensitivity to LA and has appropriate sensitivity to heparin. Patent Document 6 describes that the quality of an APTT assay reagent is stabilized by using a polymer containing a (meth)acrylic acid monomer unit having a phosphoserine residue.

[0005] Prior art literature

[0006] Patent Literature

[0007] Patent Document 1: Japanese Patent Application Laid-Open No. 59-91899

[0008] Patent Document 2: Japanese Patent Application Publication No. 2002-156379

[0009] Patent Document 3: Japanese Patent Publication No. 6-50999

[0010] Patent Document 4: Japanese Patent Application Publication No. 2013-145238

[0011] Patent Document 5: Japanese Patent Application Publication No. 2020-56578

[0012] Patent Document 6: Japanese Patent Application Publication No. 2021-181929 Summary of the invention

[0013] In coagulation tests, it is desirable to accurately detect samples with coagulation abnormalities (coagulation factor deficiency, LA positivity, coagulation inhibitor positivity, etc.). For example, it is desirable to adjust the coagulation time of these blood samples so that blood samples with coagulation abnormalities can be accurately detected based on the coagulation time.

[0014] The present invention provides a coagulation time regulator for a specimen lacking coagulation factor VIII (FVIII), coagulation factor IX (FIX) or coagulation factor XI (FXI) in an APTT assay, an APTT assay reagent, an APTT assay method using the same, and a method for regulating the coagulation time of a specimen lacking FVIII, FIX or FXI in an APTT assay.

[0015] The following aspects are provided as embodiments of the present invention.

[0016] [1] A coagulation time regulator for a blood sample deficient in coagulation factor VIII, IX or XI in an activated partial thromboplastin time measurement, comprising a polymer having 2-methacryloyloxyethylphosphocholine as a constituent unit as an active ingredient.

[0017] [2] The regulator according to [1], wherein the polymer is a copolymer of 2-methacryloyloxyethylphosphorylcholine and another monomer having a hydrophobic group, an anionic group or a cationic group.

[0018] [3] The conditioning agent according to [2], wherein the surface tension of a 0.1 wt % aqueous solution of the copolymer at 25° C. is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the above copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

[0019] [4] The regulator according to any one of [1] to [3], further comprising at least one selected from amino acids, metal salt compounds, and polysaccharides or inclusion compounds as an active ingredient.

[0020] [5] The regulator according to [4], wherein the amino acid is at least one selected from α-alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, glycylglycine, methionine, phenylalanine, proline, serine, threonine, valine and tricin.

[0021] [6] The regulator according to [4] or [5], wherein the metal salt compound is at least one selected from manganese (II) chloride, yttrium chloride, aluminum chloride, indium (III) chloride, potassium aluminum sulfate and hydrates thereof.

[0022] [7] The regulator according to any one of [4] to [6], wherein the polysaccharide or inclusion compound is at least one selected from the group consisting of dextran sulfate, cyclodextrin sulfate, and salts thereof.

[0023] [8] A reagent for measuring activated partial thromboplastin time, comprising a polymer having 2-methacryloyloxyethylphosphocholine as a constituent unit and an amino acid.

[0024] [9] The reagent according to [8], wherein the polymer is a copolymer of 2-methacryloyloxyethylphosphorylcholine and another monomer having a hydrophobic group, an anionic group, or a cationic group.

[0025]

[10] The reagent according to [9], wherein the surface tension of a 0.1 wt % aqueous solution of the copolymer at 25° C. is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the above copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

[0026]

[11] The reagent according to any one of [8] to

[10] , further comprising at least one selected from the group consisting of amino acids, metal salt compounds, and polysaccharides or inclusion compounds.

[0027]

[12] A method for measuring an activated partial thromboplastin time, comprising measuring the clotting time of a sample solution containing a blood sample and the clotting time regulator described in any one of [1] to [7] or the activated partial thromboplastin time measuring reagent described in any one of [8] to

[11] .

[0028]

[13] A method for regulating the coagulation time of a blood sample lacking coagulation factor VIII, IX or XI in an activated partial thromboplastin time assay, comprising measuring the coagulation time of a sample solution containing the blood sample and a coagulation time regulator as described in any one of [1] to [7] or an activated partial thromboplastin time assay reagent as described in any one of [8] to

[11] .

[0029]

[14] Use of a polymer having 2-methacryloyloxyethylphosphocholine as a constituent unit for regulating the coagulation time of a blood sample deficient in coagulation factor VIII, IX or XI in an activated partial thromboplastin time assay.

[0030]

[15] The use according to

[14] , wherein the polymer is a copolymer of 2-methacryloyloxyethyl phosphorylcholine and other monomers having a hydrophobic group, anionic group or cationic group.

[0031]

[16] The use according to

[15] , wherein the surface tension of a 0.1 wt % aqueous solution of the copolymer at 25° C. is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the above copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

[0032]

[17] The use according to any one of

[14] to

[16] , wherein at least one selected from amino acids, metal salt compounds, and polysaccharides or inclusion compounds is used together with the polymer.

[0033]

[18] The use according to

[17] , wherein the amino acid is at least one selected from α-alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, glycylglycine, methionine, phenylalanine, proline, serine, threonine, valine and tricin.

[0034]

[19] The use according to

[17] or

[18] , wherein the metal salt compound is at least one selected from manganese (II) chloride, yttrium chloride, aluminum chloride, indium (III) chloride, potassium aluminum sulfate and their hydrates.

[0035]

[20] The use according to any one of

[17] to

[19] , wherein the polysaccharide or inclusion compound is at least one selected from the group consisting of dextran sulfate, cyclodextrin sulfate and salts thereof.

[0036]

[21] A polymer having 2-methacryloyloxyethylphosphocholine as a constituent unit, for use in regulating the coagulation time of a blood sample deficient in coagulation factor VIII, IX or XI in an activated partial thromboplastin time measurement.

[0037]

[22] The polymer according to

[21] , which is a copolymer of 2-methacryloyloxyethyl phosphorylcholine and another monomer having a hydrophobic group, an anionic group or a cationic group.

[0038]

[23] The polymer according to

[22] , wherein the surface tension of a 0.1 wt % aqueous solution of the copolymer at 25° C. is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the above copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

[0039]

[24] The polymer according to any one of

[21] to

[23] , which is used in combination with at least one selected from the group consisting of an amino acid, a metal salt compound, and a polysaccharide or inclusion compound.

[0040]

[25] The polymer according to

[24] , wherein the amino acid is at least one selected from α-alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, glycylglycine, methionine, phenylalanine, proline, serine, threonine, valine and tricin.

[0041]

[26] The polymer according to

[24] or

[25] , wherein the metal salt compound is at least one selected from manganese (II) chloride, yttrium chloride, aluminum chloride, indium (III) chloride, potassium aluminum sulfate and hydrates thereof.

[0042]

[27] The polymer according to any one of

[24] to

[26] , wherein the polysaccharide or inclusion compound is at least one selected from the group consisting of dextran sulfate, cyclodextrin sulfate, and salts thereof.

[0043]

[28] Use of a polymer having 2-methacryloyloxyethylphosphocholine as a constituent unit for producing a coagulation time regulator for a blood sample deficient in coagulation factor VIII, IX or XI in an activated partial thromboplastin time assay.

[0044]

[29] The use according to

[28] , wherein the polymer is a copolymer of 2-methacryloyloxyethyl phosphorylcholine and other monomers having a hydrophobic group, anionic group or cationic group.

[0045]

[30] The use according to

[29] , wherein the surface tension of a 0.1 wt % aqueous solution of the copolymer at 25° C. is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the above copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

[0046]

[31] The use according to any one of

[28] to

[30] , wherein at least one selected from amino acids, metal salt compounds, and polysaccharides or inclusion compounds is used together with the above-mentioned polymer.

[0047]

[32] The use according to

[31] , wherein the amino acid is at least one selected from α-alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, glycylglycine, methionine, phenylalanine, proline, serine, threonine, valine and tricin.

[0048]

[33] The use according to

[31] or

[32] , wherein the metal salt compound is at least one selected from manganese (II) chloride, yttrium chloride, aluminum chloride, indium (III) chloride, potassium aluminum sulfate and their hydrates.

[0049]

[34] The use according to any one of

[31] to

[33] , wherein the polysaccharide or inclusion compound is at least one selected from the group consisting of dextran sulfate, cyclodextrin sulfate and salts thereof.

[0050] According to the present invention, the coagulation time of a sample lacking FVIII, FIX or FXI in the APTT assay can be adjusted, thereby accurately detecting these blood samples based on the coagulation time. DETAILED DESCRIPTION

[0051] The present invention provides a reagent for APTT assay, and a coagulation time regulator for a specimen lacking FVIII, FIX or FXI in APTT assay (hereinafter also referred to as "the coagulation time regulator of the present invention"). In addition, the present invention provides an APTT assay method using the reagent for APTT assay or the regulator, and a method for regulating the coagulation time of a specimen lacking FVIII, FIX or FXI in APTT assay.

[0052] "A specimen lacking FVIII" in this specification refers to a blood specimen lacking coagulation factor VIII (FVIII), for example, blood specimens collected from patients lacking FVIII (e.g., hemophilia A patients) and commercially available FVIII-deficient plasma. "A specimen lacking FIX" in this specification refers to a blood specimen lacking coagulation factor IX (FIX), for example, blood specimens collected from patients lacking FIX (e.g., hemophilia B patients) and commercially available FIX-deficient plasma. "A specimen lacking FXI" in this specification refers to a blood specimen lacking coagulation factor XI (FXI), for example, blood specimens collected from FXI-deficient patients and commercially available FXI-deficient plasma.

[0053] In the following description, "blood specimen" and "coagulation time" are sometimes referred to as "specimen" and "coagulation time", respectively. Therefore, in the following description, "coagulation time regulation" and "coagulation time regulator" are also referred to as "coagulation time regulation" and "coagulation time regulator", respectively.

[0054] In the APTT assay, a specimen and an APTT assay reagent are mixed to prepare a sample solution, and the coagulation reaction of the sample solution is measured. As the specimen, the plasma of the subject is preferably used. An anticoagulant commonly used in coagulation tests may be added to the specimen. For example, blood is collected using a blood collection tube filled with sodium citrate, and plasma is obtained by centrifugation. The APTT assay reagent is generally composed of a first reagent containing an activator and a second reagent containing a coagulation initiator. The specimen can be diluted with a diluent before being mixed with the APTT assay reagent as needed. The first reagent is added to the specimen, and after incubation for a specified time, the second reagent is added to initiate the coagulation reaction. The coagulation time of the sample solution after the second reagent is added is measured.

[0055] The present invention provides a coagulation time regulator for a specimen lacking FVIII, FIX or FXI in an APTT assay. In one embodiment, the coagulation time regulator of the present invention contains a polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit as an active ingredient for regulating the coagulation time of a specimen lacking FVIII, FIX or FXI. The "regulation" of the coagulation time of the specimen of the present invention includes prolongation and shortening of the coagulation time of the specimen.

[0056] The above-mentioned polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit (hereinafter sometimes referred to as a polymer) is not particularly limited as long as the constituent monomer has 2-methacryloyloxyethyl phosphorylcholine. As a polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit, it can be a polymer of 2-methacryloyloxyethyl phosphorylcholine alone, or it can be a copolymer of 2-methacryloyloxyethyl phosphorylcholine and other monomers. The other monomer may have a hydrophobic group, an anionic group or a cationic group. Or the other monomer may have both a hydrophobic group and a hydrophilic group. Preferably, the other monomer is a monomer having a hydrophobic group, an anionic group or a cationic group bonded to the 2-methacryloyloxy group, and the copolymer has a methacryloyl group in the main chain and phosphorylcholine and a hydrophobic group, anionic group or cationic group in the side chain.

[0057] The polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit can be a polymer produced according to a known synthesis method such as shown in Japanese Patent Publication No. 2004-189678. 2-methacryloyloxyethyl phosphorylcholine monomer is sold by NOF Corporation, for example, and a person skilled in the art can polymerize it with any monomer to produce a polymer having the desired properties. In addition, the polymer can use a commercially available product, such as the LIPIDURE (registered trademark) series sold by NOF Corporation. As the polymer, the LIPIDURE (registered trademark)-BL series sold as a product in the field of diagnostic drugs is preferred, but it is not limited to this. The LIPIDURE-BL series is a copolymer formed by copolymerizing various monomers with 2-methacryloyloxyethyl phosphorylcholine monomer. For example, LIPIDURE (registered trademark)-BL200 series copolymerized with a monomer having a hydrophobic group, LIPIDURE (registered trademark)-BL400 series copolymerized with a monomer having an anionic group, LIPIDURE (registered trademark)-BL500 series copolymerized with a monomer having a cationic group, LIPIDURE (registered trademark)-BL800 series copolymerized with a monomer having a hydrophobic group, LIPIDURE (registered trademark)-BL1000 series, LIPIDURE (registered trademark)-BL1100 series, LIPIDURE (registered trademark)-BL1200 series and LIPIDURE (registered trademark)-BL1300 series. More specific examples include LIPIDURE (registered trademark)-BL205, BL206 (molecular weight of about 300,000), BL405, BL503, BL504, BL802, BL1003 and BL1103.

[0058] The weight average molecular weight of the polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit is not particularly limited as long as the desired performance can be obtained. The lower limit of the weight average molecular weight of the polymer is, for example, 1000, preferably 2000, more preferably 5000, and further preferably 10000. The upper limit of the weight average molecular weight is, for example, 2000000, preferably 1000000, more preferably 700000, further preferably 100000, and further preferably 70000. The weight average molecular weight can be arbitrarily set within the range of any lower limit to any upper limit mentioned above. The weight average molecular weight of the polymer can be determined by conventional methods such as gel permeation chromatography. In addition, when the polymer is a copolymer, the constituent ratio of 2-methacryloyloxyethyl phosphorylcholine to other monomers in the copolymer varies depending on the structure of the monomers used, and generally, the content of 2-methacryloyloxyethyl phosphorylcholine in the copolymer is preferably 5 mol% or more. As for the weight average molecular weight and monomer constituent ratio of the polymer, a person skilled in the art can refer to the above-mentioned Lipidure (registered trademark)-BL series and select them according to the purpose.

[0059] The polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit preferably has a kinematic viscosity of 5 m / s at 25° C. in a 5 wt % aqueous solution. 2 More preferably, the surface tension of a 0.1 wt% aqueous solution of the polymer at 25°C is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the polymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 As for the surface tension and kinematic viscosity of the polymer, those skilled in the art can refer to the above-mentioned Lipidure (registered trademark)-BL series and select according to the purpose.

[0060] The above-mentioned polymers having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit may be used alone or in combination of any two or more.

[0061] In a preferred embodiment, the clotting time regulator of the present invention contains a copolymer having 2-methacryloyloxyethyl phosphorylcholine and other monomers as constituent units, a methacryloyl group in the main chain, phosphorylcholine and a hydrophobic group, an anionic group or a cationic group in the side chain as an active ingredient, and prolongs the clotting time of a specimen lacking FVIII, FIX or FXI in the APTT assay.

[0062] In a more preferred embodiment, the clotting time regulator of the present invention contains a copolymer as an active ingredient, and prolongs the clotting time of a specimen lacking FVIII, FIX or FXI in an APTT assay, wherein the copolymer has 2-methacryloyloxyethyl phosphorylcholine and other monomers as constituent units, and has a methacryloyl group in the main chain, phosphorylcholine and a hydrophobic group, anionic group or cationic group in the side chain, and the kinematic viscosity of a 5 wt % aqueous solution of the copolymer at 25° C. is 5 m 2 / s or less.

[0063] In a further preferred embodiment, the clotting time regulator of the present invention contains a copolymer as an active ingredient, and prolongs the clotting time of a specimen lacking FVIII, FIX or FXI in an APTT assay, wherein the copolymer has 2-methacryloyloxyethyl phosphorylcholine and other monomers as constituent units, and has a methacryloyl group in the main chain, phosphorylcholine and a hydrophobic group, anionic group or cationic group in the side chain, and the surface tension of a 0.1 wt % aqueous solution of the copolymer at 25° C. is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of its 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

[0064] The above polymer is used to adjust the clotting time of a specimen lacking FVIII, FIX or FXI in the APTT assay. The total amount of the polymer used in the APTT assay is not particularly limited as long as it is an effective amount for adjusting the clotting time of a specimen lacking FVIII, FIX or FXI in the APTT assay. For example, the total amount of the polymer used in the APTT assay is preferably 0.0005 to 0.2% by mass, more preferably 0.001 to 0.1% by mass, more preferably 0.0075 to 0.075% by mass, and more preferably 0.01 to 0.05% by mass, based on the concentration of the polymer in the sample solution for the APTT assay.

[0065] In one embodiment, the clotting time regulator of the present invention may contain at least one selected from the following amino acids, metal salt compounds, polysaccharides or inclusion compounds as an active ingredient for regulating the clotting time of a specimen lacking FVIII, FIX or FXI. For these amino acids, metal salt compounds, polysaccharides or inclusion compounds, they can be used alone as an active ingredient of the clotting time regulator, or they can be used in combination with the above-mentioned polymer as an active ingredient of the clotting time regulator.

[0066] (Amino Acid)

[0067] Examples of the amino acids include α-alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, glycylglycine, methionine, phenylalanine, proline, serine, threonine, valine, and tricin. These amino acids may be used alone or in combination of two or more.

[0068] The above amino acids are used to adjust the coagulation time of a specimen lacking FVIII, FIX or FXI in the APTT assay. It is preferably used to shorten the coagulation time of a specimen lacking FVIII, FIX or FXI. The total amount of the amino acid used in the APTT assay is not particularly limited as long as it is an effective amount for adjusting the coagulation time of a specimen lacking FVIII, FIX or FXI in the APTT assay. For example, the total amount of the amino acid used in the APTT assay is preferably 10 to 500 mM, more preferably 20 to 300 mM, more preferably 25 to 140 mM, and more preferably 30 to 135 mM, based on the concentration of the amino acid in the sample solution for the APTT assay.

[0069] (Metal Salt Compound)

[0070] As the above-mentioned metal salt compound, manganese (II) chloride, yttrium chloride, aluminum chloride, indium (III) chloride, potassium aluminum sulfate and their hydrates can be cited. As the hydrate, manganese (II) chloride tetrahydrate, yttrium chloride hexahydrate, indium (III) chloride tetrahydrate, potassium aluminum sulfate dodecahydrate, etc. can be cited. Any one of the above-mentioned metal salt compounds can be used or any two or more can be used in combination.

[0071] The above-mentioned metal salt compound is used to adjust the coagulation time of the specimen lacking FVIII, FIX or FXI in the APTT assay. The total amount of the metal salt compound used in the APTT assay is not particularly limited as long as it is an effective amount for adjusting the coagulation time of the specimen lacking FVIII, FIX or FXI in the APTT assay. For example, the total amount of the metal salt compound used in the APTT assay is preferably 0.0001 to 0.15 mg / mL, more preferably 0.001 to 0.05 mg / mL, more preferably 0.005 to 0.04 mg / mL, and more preferably 0.0075 to 0.0225 mg / mL, based on the concentration of the metal salt compound in the sample solution for the APTT assay.

[0072] (Polysaccharides or inclusion compounds)

[0073] As the above-mentioned polysaccharides or inclusion compounds, at least one polysaccharide selected from dextran, cyclodextrin, sulfated polysaccharides and their salts can be cited. Among them, sulfated polysaccharides and their salts are preferred. As sulfated polysaccharides, dextran sulfate and cyclodextrin sulfuric acid can be cited. As salts of sulfated polysaccharides, dextran sulfate sodium and cyclodextrin sulfate sodium can be cited. As cyclodextrin sulfuric acid, α-cyclodextrin sulfuric acid, β-cyclodextrin sulfuric acid, and γ-cyclodextrin sulfuric acid can be cited. As salts, sodium salts can be cited. Any one of the above-mentioned polysaccharides or inclusion compounds can be used or any two or more of them can be used in combination.

[0074] The above-mentioned polysaccharide or inclusion compound is used to adjust the coagulation time of the specimen lacking FVIII, FIX or FXI in the APTT determination. It is preferably used to prolong the coagulation time of the specimen lacking FVIII, FIX or FXI. The total amount of the polysaccharide or inclusion compound used in the APTT determination is not particularly limited as long as it is an effective amount for adjusting the coagulation time of the specimen lacking FVIII, FIX or FXI in the APTT determination. For example, the total amount of the polysaccharide or inclusion compound used in the APTT determination is preferably 0.0001 to 0.15 mg / mL, more preferably 0.001 to 0.05 mg / mL, and further preferably 0.001 to 0.01 mg / mL, based on the concentration of the polysaccharide or inclusion compound in the sample solution for the above-mentioned APTT determination.

[0075] The amino acid, metal salt compound and polysaccharide or inclusion compound mentioned above can be used as the active ingredient of the coagulation time regulator of the present invention separately or in appropriate combination. Alternatively, any one or more of the amino acid, metal salt compound and polysaccharide or inclusion compound mentioned above can be combined with any one or more of the polymer mentioned above to use as the active ingredient of the coagulation time regulator of the present invention. For example, any one or more of the polymer mentioned above can be combined with any one or more of the amino acid mentioned above to use in appropriate concentration, thereby extending or shortening the coagulation time to a desired degree.

[0076] The clotting time regulator of the present invention may further contain a buffer as an active ingredient for regulating the clotting time of a specimen lacking FVIII, FIX or FXI, in addition to the above-mentioned polymer, amino acid, metal salt compound, polysaccharide or inclusion compound.

[0077] (Buffer)

[0078] The buffering agent may be at least one selected from PIPES, ACES, HEPES, TAPSO, POPSO, EPPS and CHES. The total amount of the buffering agent used in the APTT assay is preferably 0.1 to 1000 mM, more preferably 0.5 to 500 mM, further preferably 1 to 200 mM, and further preferably 10 to 100 mM, based on the concentration in the sample solution.

[0079] The buffers listed above may be used alone or in combination of any two or more.

[0080] The clotting time regulator of the present invention is used to regulate the clotting time of a specimen lacking FVIII, FIX or FXI in APTT assay. The clotting time regulator of the present invention can be added alone to the above-mentioned specimen, diluent or sample solution for APTT assay, or the APTT assay reagent containing the clotting time regulator of the present invention can be added to the above-mentioned specimen, diluent or sample solution.

[0081] Therefore, the present invention provides an APTT assay reagent comprising the above-mentioned polymer having 2-methacryloyloxyethylphosphorylcholine as a constituent unit, an amino acid, a metal salt compound, a polysaccharide, or an inclusion compound.

[0082] In one embodiment, the APTT assay reagent of the present invention contains at least one selected from the above-mentioned polymer, amino acid, metal salt compound, polysaccharide or inclusion compound. Preferably, the APTT assay reagent of the present invention contains the above-mentioned polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit. In one embodiment, the APTT assay reagent of the present invention contains the above-mentioned polymer and at least one selected from the above-mentioned amino acid, metal salt compound, polysaccharide or inclusion compound. In one embodiment, the APTT assay reagent of the present invention contains the above-mentioned polymer and the above-mentioned amino acid.

[0083] In one embodiment, the APTT assay reagent of the present invention may contain the above-mentioned buffer in addition to at least one selected from the above-mentioned polymer, amino acid, metal salt compound, and polysaccharide or inclusion compound.

[0084] The APTT assay reagent of the present invention preferably contains the above-mentioned polymer and amino acid, and, if necessary, contains at least one selected from the group consisting of the above-mentioned metal salt compound, polysaccharide or inclusion compound, and a buffer.

[0085] The APTT assay reagent of the present invention is preferably a kit comprising a first reagent containing an activator and a second reagent containing a coagulation initiator. In the APTT assay reagent, at least one selected from the above-mentioned polymer, amino acid, metal salt compound, and polysaccharide or inclusion compound is preferably contained in the first reagent, but may be contained in the kit separately from the first reagent and the second reagent.

[0086] The content of the polymer, amino acid, metal salt compound, polysaccharide or inclusion compound in the APTT assay reagent of the present invention can be adjusted in such a manner that the concentration of the polymer, amino acid, metal salt compound, polysaccharide or inclusion compound contained in the sample solution obtained by mixing the APTT assay reagent with the specimen is within the above-mentioned effective amount range for adjusting the clotting time of the specimen lacking FVIII, FIX or FXI in the APTT assay. The same applies to the content of the buffer in the APTT assay reagent.

[0087] The APTT assay reagent provided by the present invention may contain, in addition to at least one selected from the above-mentioned polymers, amino acids, metal salt compounds, polysaccharides or inclusion compounds, various components required for activation or coagulation of the blood sample, etc. For example, in the APTT assay reagent, the first reagent may contain a buffer, an activator, a phospholipid, etc.

[0088] The buffer can be appropriately selected and used in the range of pH 4 to 9, preferably pH 6 to 8. Examples of the buffer include the above-mentioned buffer, or amino acids, citric acid, phosphoric acid, acetic acid, imidazole, barbital, GTA, etc. The amino acid as the active ingredient of the above-mentioned coagulation time regulator of the present invention can also be used as a buffer. Any one of these buffers can be used or any two or more can be used in combination. The amount of the buffer used is not particularly limited as long as it is an amount that exerts buffering capacity, and is preferably 0.1 to 1000mM, more preferably 0.5 to 500mM, more preferably 1 to 200mM, and more preferably 10 to 100mM in terms of concentration in the reagent.

[0089] Examples of the activator include ellagic acid, kaolin, diatomaceous earth, colloidal silica, polyphenol compounds, anhydrous silicic acid, and metal ions. Examples of the metal ions include Zn 2+ , Mn 2+ , Cu 2+ , Fe 2+ 、Al 3+ Etc. The metal ion may be contained in the above reagent in the form of a salt. The metal salt compound which is the active ingredient of the above-mentioned coagulation time regulator of the present invention may also be used as an activator. Any one of the above-mentioned activators may be used or any two or more of them may be used in combination. The amount of the activator used is not particularly limited, and is preferably 0.001 to 2 mg / mL, more preferably 0.001 to 0.5 mg / mL, further preferably 0.001 to 0.1 mg / mL, and further preferably 0.005 to 0.05 mg / mL, based on the concentration in the reagent.

[0090] As examples of the above-mentioned phospholipids, natural or synthetic phospholipids can be cited. As the natural phospholipids, phospholipids from natural substances, such as rabbit brain, bovine brain, human placenta, soybean, egg yolk, etc. can be cited. As the synthetic phospholipids, for example, phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidic acid (PA), phosphatidylinositol (PI), lysophosphatidylcholine (LPC), sphingomyelin (SM), cardiolipin, etc. can be cited. Any one of these phospholipids can be used or any two or more can be used in combination. For example, a variety of phospholipids can be mixed and used as liposomes. The amount of the phospholipid used is not particularly limited, and is preferably 0.005 to 2 mM, more preferably 0.02 to 0.5 mM, in terms of the concentration in the reagent. In the APTT assay reagent, the phospholipids as platelet substitutes are added as components required for the coagulation reaction. In the intrinsic coagulation reaction, phospholipids form a complex with activated factors IX and VIII, which forms fibrin through a common factor system (factors V, II and I) following activation of factor X.

[0091] The second reagent of the APTT assay reagent may contain a component that triggers blood coagulation, such as calcium ions. As calcium ions, water-soluble calcium compounds such as calcium chloride, calcium lactate, calcium gluconate, calcium glucuronate, and calcium tartrate can be used. These calcium compounds can be used alone or in combination of any two or more. The amount of the calcium compound used is not particularly limited, and is preferably 5 mM to 100 mM, more preferably 10 mM to 50 mM, based on the concentration in the reagent.

[0092] The first reagent and the second reagent may further contain additives for improving the preservation or stability of the reagent. Examples of such additives include preservatives, antioxidants, dispersants, stabilizers, etc. Examples of the preservative include ciprofloxacin, propionic acid, sodium benzoate, sodium azide, a mixture of 2-methyl-1,2-thiazol-3(2H)-one and 5-chloro-2-methyl-1,2-thiazol-3(2H)-one (e.g., Proclin 300), etc. Examples of the antioxidant include citric acid, butylated hydroxyanisole, etc. Examples of the dispersant include phenol, collagen peptide, etc. Examples of the stabilizer include polyethylene glycol, polyvinyl pyrrolidone, high molecular weight polysaccharides such as dextran or polysucrose (e.g., Ficoll), salts such as sodium chloride, amino acids, sugars, etc.

[0093] The coagulation time regulator and the APTT assay reagent of the present invention can be in the form of a liquid or its frozen product, or can also be in a dry form. The dry form reagent can be dissolved in water, a buffer solution, etc. to prepare a liquid reagent when used. The concentrations of the above-mentioned various components are expressed as concentrations in the liquid reagent.

[0094] The present invention provides a method for measuring a coagulation reaction (APTT measurement) using the clotting time regulator or APTT measurement reagent of the present invention. The APTT measurement method of the present invention can adjust the clotting time of a specimen lacking FVIII, FIX or FXI by using the clotting time regulator or APTT measurement reagent of the present invention.

[0095] The APTT determination method of the present invention can be implemented according to conventional methods except using the clotting time regulator or APTT determination reagent of the present invention. In a preferred embodiment, the first reagent containing the clotting time regulator of the present invention is mixed in the specimen diluted as needed, and the obtained mixed solution is heated. The heating conditions are, for example, 30°C to 40°C, preferably 35°C to 39°C. Thereafter, the second reagent is added to the mixed solution to induce a coagulation reaction. The coagulation reaction of the mixed solution (sample solution) after the second reagent is added is measured. The measurement can be performed using an optical method for measuring the amount of scattered light, transmittance or absorbance of the sample solution, or a mechanical method for measuring the viscosity of the sample solution. The temperature of the sample solution in the measurement is, for example, 30°C to 40°C, preferably 35°C to 39°C. The APTT determination reagent is preferably added to the specimen at a concentration such that the clotting time (APTT) of normal plasma falls within the range of 15 to 80 seconds, preferably 15 to 60 seconds, and more preferably 20 to 50 seconds.

[0096] The reaction start time of the coagulation reaction can be typically defined as the time when the second reagent is mixed in the specimen to induce the coagulation reaction. Alternatively, other times can be used as the reaction start time. The time for continuously measuring the coagulation reaction can be, for example, from the time when the specimen and the second reagent are mixed for several tens of seconds to about 7 minutes. The measurement time can be an arbitrarily determined fixed value, or it can be until the time when the coagulation reaction of each specimen is detected to be finished. The measurement of the progress of the coagulation reaction (for example, the measurement of the amount of scattered light) can be repeatedly performed at specified intervals within the measurement time. For example, it is sufficient to measure at intervals of 0.1 seconds. The clotting time (APTT) can be determined according to a known method based on the measured coagulation reaction.

[0097] The series of operations in the APTT assay method of the present invention can be performed using an automatic analyzer. Alternatively, a portion of the operations can be performed manually. For example, the specimen can be prepared by a person, and the subsequent operations can be performed by an automatic analyzer.

[0098] Example

[0099] The present invention will be described in further detail below with reference to Examples, but the present invention is not limited to these Examples.

[0100] Test 1

[0101] 1) Specimen

[0102] The following specimens were used.

[0103] FVIII-deficient specimens (specimens from patients with FVIII deficiency, N=5)

[0104] FIX-deficient specimens (specimens from FIX-deficient patients, N=5)

[0105] FXI-deficient specimens (specimens from patients with FXI deficiency, N=4)

[0106] 2) APTT assay reagent

[0107] 2-1) Basic formula

[0108] Reagent 1 (R1)

[0109]

[0110] Reagent 2 (R2)

[0111] Element concentration

[0112] Calcium chloride 20 mM

[0113] Sodium azide 0.05 mass%

[0114] *1) Prepared according to the method described in International Publication No. 2003 / 015753.

[0115] 2-2) Test reagents

[0116] Test reagents 1-1 to 1-20: A polymer having 2-methacryloyloxyethyl phosphorylcholine as a constituent unit (LIPIDURE (registered trademark)-BL series; BL205, BL206, BL405, BL503, BL504, BL802, BL1003 or BL1103) was added to the basic formulation R1 under the conditions described in Table 1. As a control, a reagent of the basic formulation was used.

[0117] Test reagents 2-1 to 2-14: In the basic formulation R1, amino acids were added in place of glycine under the conditions described in Table 2. As a control, R1 containing lysine in place of glycine was prepared.

[0118] Test reagents 3-1 to 3-11: In the basic formula R1, instead of 0.015 mg / mL copper sulfate pentahydrate, manganese (II) chloride tetrahydrate, yttrium chloride hexahydrate, aluminum chloride (anhydrous), indium (III) chloride tetrahydrate, or potassium aluminum sulfate dodecahydrate was added under the conditions described in Table 3. As a control, the reagent of the basic formula was used.

[0119] Test reagents 4-1 to 4-6: Dextran sulfate sodium or β-cyclodextrin sulfate sodium was added to the basic formulation R1 under the conditions described in Table 4. As a control, a reagent of the basic formulation was used.

[0120] 2-3) Preparation of R1 and R2

[0121] The above components were mixed to give predetermined concentrations to prepare each test reagent and control R1 and R2. The mixing of R1 was performed under ice cooling.

[0122] 3) Determination of coagulation time

[0123] After heating 50 μL of the specimen at 37°C in a cuvette for 45 seconds, 50 μL of the first reagent (R1) is added and further heated for 171 seconds. 50 μL of the second reagent (R2) is added to the heated mixture to initiate a coagulation reaction. The reaction is carried out at 37°C. The specimen (sample solution) to which R2 is added is irradiated with light of a wavelength of 660 nm, and the optical change (the change in scattered light intensity) caused by the coagulation reaction is measured. The coagulation time is calculated from the measured coagulation reaction. The coagulation time is measured using a coagulation automatic analyzer CP3000 (manufactured by Sekisui Medical Co., Ltd.). The change rate (%) of the coagulation time (APTT) using the test reagent relative to the coagulation time (APTT) using the control (basic formula) is calculated [change rate (%) = (APTT using the test reagent) / APTT using the control × 100]. A change rate of less than 100% indicates a shortened coagulation time, and a change rate of more than 100% indicates a prolonged coagulation time.

[0124] The results (average values ​​of the change rates of each sample type) are shown in Tables 5 to 8. Using the test reagents to which the compounds described in Tables 1 to 4 were added, the clotting time of the samples lacking FVIII, FIX, and FXI was prolonged or shortened compared with the control.

[0125] [Table 1]

[0126]

[0127] [Table 2]

[0128]

[0129] [Table 3]

[0130]

[0131] [Table 4]

[0132]

[0133] [Table 5]

[0134]

[0135] [Table 6]

[0136]

[0137] [Table 7]

[0138]

[0139] [Table 8]

[0140]

[0141] Test 2

[0142] 1) Specimen

[0143] The following specimens were used.

[0144] Normal specimens (mixed plasma from normal volunteers, N=3)

[0145] Samples lacking FVIII (samples from patients with FVIII deficiency, N=5) Samples lacking FIX (samples from patients with FVIII deficiency, N=5)

[0146] FXI-deficient specimens (specimens from patients with FVIII deficiency, N=3)

[0147] 2) APTT assay reagent

[0148] 2-1) Basic formula

[0149] The same formulation as in Experiment 1, except that the concentration of copper sulfate pentahydrate in the first reagent (R1) was changed within the range of 0.0075 to 0.0225 mg / mL.

[0150] 2-2) Test reagents

[0151] Test reagent 5-1: LIPIDURE (registered trademark)-BL205 was added to the basic formulation R1 (copper sulfate pentahydrate concentration: 0.0075 mg / mL) under the conditions described in Table 9. The case where LIPIDURE (registered trademark) was not added was used as a control.

[0152] Test reagent 6-1: LIPIDURE (registered trademark)-BL205 was added to the basic formulation R1 (copper sulfate pentahydrate concentration: 0.0225 mg / mL) under the conditions described in Table 10. The case where LIPIDURE (registered trademark) was not added was used as a control.

[0153] Test reagent 7-1: The amino acid (glycine) in the basic formula R1 (copper sulfate pentahydrate concentration: 0.015 mg / mL) was changed to the conditions described in Table 12, and LIPIDURE (registered trademark)-BL205 was further added. The case without adding LIPIDURE (registered trademark) was used as a control.

[0154] Test reagent 2-15: The amino acid (glycine) in the basic formula R1 (copper sulfate pentahydrate concentration: 0.015 mg / mL) was changed to lysine, and LIPIDURE (registered trademark)-BL205 was further added. The case without adding LIPIDURE (registered trademark) (Control 2 in Table 2) was used as a control.

[0155] Test reagent 8-1: The copper sulfate pentahydrate in the basic formula R1 was changed to the conditions described in Table 13, and LIPIDURE (registered trademark)-BL205 was further added. The case where LIPIDURE (registered trademark) was not added was used as a control.

[0156] Test reagent 9-1: As shown in Table 13, zinc chloride in test reagent 8-1 was replaced with manganese (II) chloride tetrahydrate. Test reagent 8-1 was used as a control.

[0157] 2-3) Reagent preparation and clotting time determination

[0158] R1 and R2 were prepared by the same procedure as in Test 1, and the clotting time (APTT) was measured to determine the change rate (%) of the APTT of the test reagent relative to the control. In addition, for each test reagent, the change rate of the APTT of the LA-positive specimen or the specimen containing heparin relative to the APTT of the normal specimen was determined [change rate relative to the normal specimen (%) = (APTT of the specimen lacking coagulation factors) / (APTT of the normal specimen) × 100].

[0159] The results are shown in Tables 9 to 13. As shown in Tables 9 to 10, even when the concentration of the metal salt compound (copper sulfate pentahydrate) is different, the coagulation time of the specimen lacking FVIII, FIX and FXI is prolonged compared with the control by adding a polymer (LIPIDURE (registered trademark)). Among them, as shown in Table 11, the effect of extending the coagulation time of the specimen lacking FVIII, FIX and FXI using the test reagent is weaker than that of the normal specimen. Specifically, the APTT variation rate relative to the normal specimen is 95-100% in the specimen lacking FVIII, 94-99% in the specimen lacking FIX, and 93-102% in the specimen lacking FXI. In addition, according to the results of Tables 12 to 13, it is shown that reagents with different types of amino acids or metal salt compounds combined with polymers all play the same effect of extending the coagulation time of the specimen lacking FVIII, FIX and FXI.

[0160] [Table 9]

[0161]

[0162] [Table 10]

[0163]

[0164] [Table 11]

[0165]

[0166] [Table 12]

[0167]

[0168] [Table 13]

[0169]

Claims

1. A coagulation time regulator for a blood sample deficient in coagulation factor VIII, IX or XI in an activated partial thromboplastin time measurement, comprising a polymer having 2-methacryloyloxyethylphosphocholine as a constituent unit as an active ingredient.

2. The regulator according to claim 1, wherein The polymer is a copolymer of 2-methacryloyloxyethyl phosphorylcholine and other monomers having a hydrophobic group, an anionic group or a cationic group.

3. The regulator according to claim 2, wherein The surface tension of a 0.1 wt% aqueous solution of the copolymer at 25°C is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

4. The regulator according to claim 1, wherein Furthermore, at least one selected from amino acids, metal salt compounds, and polysaccharides or inclusion compounds is used as an active ingredient.

5. The regulator according to claim 4, wherein The amino acid is at least one selected from α-alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, glycylglycine, methionine, phenylalanine, proline, serine, threonine, valine and tricin.

6. The regulator according to claim 4, wherein The metal salt compound is at least one selected from the group consisting of manganese (II) chloride, yttrium chloride, aluminum chloride, indium (III) chloride, potassium aluminum sulfate, and hydrates thereof.

7. The regulator according to claim 4, wherein The polysaccharide or inclusion compound is at least one selected from the group consisting of dextran sulfate, cyclodextrin sulfate, and salts thereof.

8. A reagent for measuring activated partial thromboplastin time, comprising a polymer having 2-methacryloyloxyethylphosphocholine as a constituent unit and an amino acid.

9. The reagent according to claim 8, wherein The polymer is a copolymer of 2-methacryloyloxyethyl phosphorylcholine and other monomers having a hydrophobic group, an anionic group or a cationic group.

10. The reagent according to claim 9, wherein The surface tension of a 0.1 wt% aqueous solution of the copolymer at 25°C is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

11. The reagent according to claim 8, wherein It further contains at least one selected from the group consisting of amino acids, metal salt compounds, and polysaccharides or inclusion compounds.

12. A method for measuring an activated partial thromboplastin time, comprising measuring the coagulation time of a sample solution containing a blood sample and the coagulation time regulator according to any one of claims 1 to 7 or the activated partial thromboplastin time measuring reagent according to any one of claims 8 to 11.

13. A method for regulating the coagulation time of a blood sample lacking coagulation factor VIII, IX or XI in an activated partial thromboplastin time measurement, comprising measuring the coagulation time of a sample solution containing the blood sample and the coagulation time regulator according to any one of claims 1 to 7 or the activated partial thromboplastin time measurement reagent according to any one of claims 8 to 11.

14. Use of a polymer having 2-methacryloyloxyethylphosphocholine as a constituent unit for regulating the coagulation time of a blood sample deficient in coagulation factor VIII, IX or XI in an activated partial thromboplastin time assay.

15. The use according to claim 14, wherein: The polymer is a copolymer of 2-methacryloyloxyethyl phosphorylcholine and other monomers having a hydrophobic group, an anionic group or a cationic group.

16. The use according to claim 15, wherein: The surface tension of a 0.1 wt% aqueous solution of the copolymer at 25°C is 37 to 60×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 0.5 to 3 m 2 / s; or the surface tension of a 0.1 wt% aqueous solution of the copolymer at 25°C is 70 to 75×10 -3 N / m, and the kinematic viscosity of a 5 wt% aqueous solution at 25°C is 2 to 5 m 2 / s.

17. The use according to claim 14, wherein: At least one selected from the group consisting of amino acids, metal salt compounds, and polysaccharides or inclusion compounds is used together with the polymer.

18. The use according to claim 17, wherein: The amino acid is at least one selected from α-alanine, arginine, asparagine, aspartic acid, glutamic acid, glycine, glycylglycine, methionine, phenylalanine, proline, serine, threonine, valine and tricin.

19. The use according to claim 17, wherein: The metal salt compound is at least one selected from the group consisting of manganese (II) chloride, yttrium chloride, aluminum chloride, indium (III) chloride, potassium aluminum sulfate, and hydrates thereof.

20. The use according to claim 17, wherein: The polysaccharide or inclusion compound is at least one selected from the group consisting of dextran sulfate, cyclodextrin sulfate, and salts thereof.

Images