WO2015194165A1 - Accélérateur de coagulation du sang, et agent pour tester la fonction de caillots sanguins à l'aide de celui-ci - Google Patents

Accélérateur de coagulation du sang, et agent pour tester la fonction de caillots sanguins à l'aide de celui-ci Download PDF

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WO2015194165A1
WO2015194165A1 PCT/JP2015/003003 JP2015003003W WO2015194165A1 WO 2015194165 A1 WO2015194165 A1 WO 2015194165A1 JP 2015003003 W JP2015003003 W JP 2015003003W WO 2015194165 A1 WO2015194165 A1 WO 2015194165A1
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blood coagulation
particles
reagent
acid
blood
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PCT/JP2015/003003
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English (en)
Japanese (ja)
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浩平 白石
こずえ 岡野
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学校法人近畿大学
国立大学法人山口大学
日油株式会社
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Priority to JP2016529048A priority Critical patent/JP6692023B2/ja
Publication of WO2015194165A1 publication Critical patent/WO2015194165A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/86—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood coagulating time or factors, or their receptors

Definitions

  • the present disclosure relates to a blood coagulation promoter and a blood coagulation function test drug using the same.
  • Blood coagulation is a complex mechanism involving various factors, and whether these mechanisms are functioning normally is determined by screening tests prior to surgery and screening tests for abnormal blood coagulation such as hemophilia. Etc. are important.
  • Activating partial thromboplastin time is a test of the intrinsic coagulation system with improved sensitivity and accuracy.
  • blood coagulation reaction is promoted by adding a phospholipid called partial thromboplastin fraction together with calcium and an active agent composed of ellagic acid and the like.
  • the examination of the exogenous coagulation system includes prothrombin time (PT) using a protein called tissue thromboplastin and calcium.
  • composition of the test agent for the purpose of improving the accuracy or sensitivity of the test relating to blood coagulation (see, for example, Patent Document 1).
  • test drugs basically use components derived from living bodies as drugs that promote blood coagulation, which is the main component. For this reason, the dispersion
  • An object of the present disclosure is to realize a blood coagulation promoter exhibiting a stable blood coagulation action and a blood coagulation function test drug using the same.
  • the first aspect of the blood coagulation promoter of the present disclosure includes particles having carboxyl groups oriented on the surface.
  • the surface density of carboxyl groups in the particles 0.5 [mu] mol / m 2 or more, can be 50 [mu] mol / m 2 or less.
  • the particles can be a resin.
  • the resin may include a monomer unit derived from a monomer having a carboxyl group.
  • the monomer having a carboxyl group can be at least one of itaconic acid, acrylic acid and methacrylic acid.
  • the resin can be a polymer containing 10 mol% or more of monomer units derived from a monomer having a carboxyl group.
  • the particles may be cellulose having a carboxyl group.
  • the particles may have an amino group on the surface.
  • the second aspect of the blood coagulation promoter contains sponge-like titanium oxide particles.
  • the particles of sponge-like titanium oxide, surface area 50 m 2 / g or more may be 1000 m 2 / g or less.
  • the particles may have an average particle diameter of 10 nm or more and 10 ⁇ m or less.
  • the blood coagulation function test drug contains at least one of the blood coagulation promoters of the first aspect and the second aspect of the present disclosure.
  • a stable blood coagulation action can be realized, and a stable blood coagulation function test drug can be realized.
  • FIG. 1 (a) and 1 (b) are diagrams for explaining the promotion of blood coagulation by a surface in which carboxyl groups are oriented
  • FIG. 1 (a) is a diagram showing a surface in which carboxyl groups are oriented
  • 1 (b) is a view showing a surface in which carboxyl groups are not oriented.
  • FIG. 2 is an electron micrograph of plasma coagulated with particles having carboxyl groups oriented on the surface.
  • FIG. 3 is an electron micrograph of plasma coagulated with particles having carboxyl groups oriented on the surface.
  • FIG. 4 is an electron micrograph of plasma coagulated with sponge-like titanium oxide particles.
  • FIG. 5 is an electron micrograph of plasma coagulated with particles having PMPC fixed on the surface.
  • FIG. 6 is an electron micrograph of plasma coagulated by the addition of calcium.
  • the blood coagulation promoter of the present disclosure includes particles having carboxyl groups oriented on the surface.
  • a blood coagulation promoter is a drug that promotes the coagulation reaction more than calcium alone by adding it together with calcium to whole blood or plasma containing an anticoagulant having a decalcification action.
  • the particle having a carboxyl group oriented on the surface is a particle in which the carboxyl group is oriented outward and the carboxyl group is exposed on the particle surface.
  • Surface density of carboxyl groups is not particularly limited, 0.5 [mu] mol / m 2 or more, preferably 1.0 [mu] mol / m 2 or more, more preferably 5.0 ⁇ mol / m 2 or more, 50 [mu] mol / m 2 or less, preferably 25 ⁇ mol / M 2 or less, more preferably 10 ⁇ mol / m 2 or less.
  • the amount of the carboxyl group can be determined by an electric conductivity titration method.
  • the surface density of the carboxyl group can be obtained by dividing the amount of the carboxyl group by the value of the specific surface area measured by the BET (Brunauer, Emmet and Teller) method or the like.
  • the average particle diameter of the particles is not particularly limited, but may be 0.01 ⁇ m or more, preferably 0.05 ⁇ m or more, more preferably 0.1 ⁇ m or more, 10 ⁇ m or less, preferably 5 ⁇ m or less, more preferably 1 ⁇ m or less. .
  • the average particle diameter can be measured by a laser diffraction particle size distribution measuring device or the like.
  • the particles may be spherical, flaky, scaly, or nanofibers. Further, it may be solid, hollow, or a porous shape having pores. The surface of the particles may be smooth or uneven.
  • the particles may be any particles as long as they can stably obtain the same characteristics.
  • industrially produced particles can be used. It may be derived from plants, animals or microorganisms as long as the same properties can be obtained stably.
  • the particles may be an organic substance such as a resin or an inorganic substance such as silica.
  • the particles may be solid or may be a material having a certain degree of fluidity such as hydrogel or liposome.
  • Examples of industrially produced particles include synthetic polymers.
  • the synthetic polymer can be, for example, a polymer including a monomer unit having a carboxyl group.
  • a polymer including monomer units derived from monomers such as acrylic acid, methacrylic acid, or itaconic acid may be used.
  • the polymer may be a homopolymer composed of a monomer having a carboxyl group, or may be a binary or higher copolymer.
  • the polymer may be cross-linked or non-cross-linked as long as it can be made into particles having a predetermined particle size.
  • the polymer may contain any other monomer unit as long as it contains a monomer unit having a carboxyl group.
  • the monomer unit derived from styrene may be included. It may contain a monomer unit derived from a hydrophobic monomer such as acrylic acid or methacrylic acid ester such as methyl acrylate or methyl methacrylate.
  • a hydrophobic monomer such as acrylic acid or methacrylic acid ester such as methyl acrylate or methyl methacrylate.
  • Contains monomer units derived from hydrophilic monomers such as N-vinylpyrrolidone, 2-methacryloyloxyethyl phosphorylcholine (MPC), 2-dimethylaminoethyl methacrylate, 2-hydroxyethyl methacrylate, allylamine, or polyethylene glycol macromonomer You may go out.
  • the monomer unit which has a sulfonic acid group or a phosphoric acid group etc. may be included.
  • a monomer unit having a functional group having a positive charge such as an amino group may be included instead of the functional group having a negative charge.
  • the content of the monomer unit having a carboxyl group in the polymer is not particularly limited, but can be 10 mol% or more, preferably 20 mol% or more, more preferably 50 mol% or more. Since the aggregation promoting effect is considered to be higher as the surface density of the carboxyl group is higher, a polymer in which all the monomer units are monomer units having a carboxyl group may be used. Further, the content of the monomer unit having a carboxyl group in the polymer may be 95 mol% or less, 90% or less, or 80% or less. By adjusting the content of the monomer unit having a carboxyl group, the density of the carboxyl group on the surface of the particle can be easily adjusted.
  • a carboxyl group may be introduced into the particle after forming the particle.
  • the amino group is reacted with a dicarboxylic acid such as oxalic acid or a tricarboxylic acid such as citric acid to thereby form particles having carboxyl groups oriented on the surface. It may be formed. In this case, amino groups may remain on the surface of the particles.
  • maleic anhydride can be hydrolyzed to introduce a carboxyl group.
  • Natural polymers or modified ones may be used instead of synthetic polymers.
  • cellulose introduced with a carboxyl group can be used.
  • the cellulose for example, cellulose nanofibers crushed so as to have a length of several ⁇ m to several tens of ⁇ m can be used.
  • Cellulose can be crushed using physical methods, chemical methods, or both.
  • Introduction of a carboxyl group into cellulose can be performed, for example, by oxidizing a hydroxyl group. Of these, oxidation using 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO) is preferable.
  • the introduction of the carboxyl group may be performed after pulverization of cellulose or before pulverization. Moreover, you may perform crushing and introduction
  • the concentration in the blood of particles having carboxyl groups oriented on the surface is not particularly limited, but is 0.5 ⁇ g / mL or more, preferably 5 ⁇ g / mL or more, more preferably 50 ⁇ g / mL or more, It can be 1000 ⁇ g / mL or less, preferably 500 ⁇ g / mL or less, more preferably 250 ⁇ g / mL or less.
  • the concentration of calcium in the blood is 0.5 mmol / L or more, preferably 1 mmol / L or more, more preferably 5 mmol / L or more, 100 mmol / L or less, preferably 50 mmol / L or less, more preferably 25 mol. / L or less may be added.
  • Calcium can be added to blood as an inorganic or organic salt.
  • Blood can be coagulated, whether it is whole blood or plasma. Blood can be coagulated even if it contains an anticoagulant having a decalcifying action such as citrate or ethylenediaminetetraacetic acid.
  • an anticoagulant having a decalcifying action such as citrate or ethylenediaminetetraacetic acid.
  • the blood coagulation promoter containing particles having carboxyl groups oriented on the surface can be used as a blood coagulation function test agent used for hemostasis function test and the like. Similar to the prothrombin time (PT) reagent or the activated partial thromboplastin time (APTT) reagent, the function of the coagulation system can be examined by measuring the coagulation time.
  • PT prothrombin time
  • APTT activated partial thromboplastin time
  • the first reagent containing the blood coagulation promoter and the sample are mixed, then the second reagent containing calcium is mixed, and the coagulation time until fibrin is precipitated is measured. do it.
  • the time until fibrin precipitates can be measured visually or by a change in absorbance.
  • the first reagent can be a suspension containing particles in which carboxyl groups are oriented.
  • grains in which the carboxyl group in the 1st reagent was orientated should just be a density
  • the concentration of the particles in which the carboxyl group is oriented in the first reagent is 1.5 ⁇ g / mL or more, preferably 10 ⁇ g / mL or more, more preferably 50 ⁇ g / mL or more, particularly preferably 150 ⁇ g / mL or more, 3000 ⁇ g / mL.
  • the second reagent is, for example, calcium 1.5 mmol / L or more, preferably 3 mmol / mL or more, more preferably 15 mmol / L or more, 300 mmol / L or less, preferably 150 mmol / mL or less, more preferably 75 mmol / L or less. It can be set as the solution containing.
  • a first reagent containing calcium may be prepared.
  • the coagulation time may be measured by mixing the first reagent and the specimen.
  • the particles having a carboxyl group oriented on the surface have a coagulation promoting function similar to that of tissue thromboplastin in PT reagent or partial thromboplastin fraction and activator in APTT reagent.
  • the first reagent does not contain tissue thromboplastin or partial thromboplastin fractions but promotes blood coagulation.
  • Platelets activated by bleeding or the like and exposed to phosphatidylserine promote blood coagulation.
  • the particles having carboxyl groups oriented on the surface may promote the blood coagulation reaction by a mechanism similar to that of activated platelets exposed to phosphatidylserine.
  • phosphatidylserine On the surface of the activated platelet cell membrane, phosphatidylserine is considered to be oriented and exposed outward.
  • a weakly acidic substance or a complex thereof may be added to the blood coagulation function test drug.
  • one or more of the following drugs can be added.
  • Sugar derivatives such as glucuronic acid, uronic acid, mannuronic acid, aldonic acid, aldaric acid, hyaluronic acid, or carboxylic acid-containing sugar chains.
  • a dicarboxylic acid such as oxalic acid or malonic acid, an acidic protein, or an acidic peptide, a monoanion, or a polyanion.
  • Nucleic acids such as deoxyribonucleic acid or ribonucleic acid, peptide nucleic acids, fumaric acid, maleic acid, phthalic acid, isophthalic acid, terephthalic acid, pyromellitic acid, citric acid, malic acid, or polybasic acids such as ethylenediaminetetraacetic acid.
  • Phospholipids such as phosphatidylinositol or phosphatidylserine.
  • Acidic amino acids such as aspartic acid or glutamic acid.
  • Blood coagulation factors such as tissue factor or thrombin.
  • These agents may be added in a state of being bound to a carrier.
  • a carrier for example, one or more of the following can be used.
  • Metal particles such as gold or silver, inorganic particles such as kaolin, monolithic silica or colloidal silica, carbon particles such as carbon black, fullerene and fullerene nanotubes, inclusion compounds such as cyclodextrin or calixarene, pullulan, mannan, dextran or Polysaccharides such as amylopectin, liposomes, vesicles, calcium phosphate, hydroxyapatite, magnetic particles, multifunctional envelope-type nanostructures, polymer micelles, polyion complexes, polycyclic aromatic compounds, viruses, cells, microorganisms, brain sulfide, Or fibrin etc.
  • drugs or drugs supported on a carrier can be added to the first reagent and mixed with the specimen together with the first reagent. Moreover, you may mix with a test substance separately from a 1st reagent. Further, it may be added to the second reagent.
  • the first reagent and the second reagent may further contain a component contained in a general test agent such as a preservative.
  • Spongy titanium oxide particles can be used as a blood coagulation promoter instead of particles having carboxyl groups oriented on the surface.
  • Sponge-like titanium oxide particles are titanium oxide particles having a three-dimensional network structure.
  • Sponge-like titanium oxide particles have a large number of Lewis acid sites on the surface. This Lewis acid site is considered to act in the same manner as the carboxyl group.
  • the titanium oxide may be a rutile type or an anatase type.
  • Sponge-like titanium oxide particles are not particularly limited, but the surface area is 50 m 2 / g or more, preferably 100 m 2 / g or more, more preferably 200 m 2 / g or more, 1000 m 2 / g or less, preferably 800 m 2 / g. More preferably 500 m 2 / g or less.
  • the average particle diameter of the sponge-like titanium oxide particles is not particularly limited, but is 0.01 ⁇ m or more, preferably 0.05 ⁇ m or more, more preferably 0.1 ⁇ m or more, 100 ⁇ m or less, similarly to the particles having a carboxyl group on the surface. Preferably it is 50 micrometers or less, More preferably, it can be 10 micrometers or less.
  • the sponge-like titanium oxide particles are not particularly limited, and those formed by entanglement of fibrous titanium oxide so as to form pores of about several nm to 100 nm can be used. Moreover, what was formed by the hydrothermal synthesis method etc. can be used.
  • the concentration of the sponge-like titanium oxide particles in the final blood is 0.2 ⁇ g / mL or more, Preferably, it can be 1 ⁇ g / mL or more, more preferably 2 ⁇ g / mL or more, 400 ⁇ g / mL or less, preferably 100 ⁇ g / mL or less, more preferably 40 ⁇ g / mL or less.
  • the concentration of calcium in the blood is 0.5 mmol / L or more, preferably 1 mmol / L or more, more preferably 5 mmol / L or more, 100 mmol / L or less, preferably 50 mmol / L or less, more preferably 25 mol / L or less. can do.
  • the concentration of the sponge-like titanium oxide particles contained in the first reagent is 0.6 ⁇ g / mL or more, preferably 3 ⁇ g / mL or more, more preferably 6 ⁇ g / mL or more, 1200 ⁇ g / mL or less, preferably 300 ⁇ g / mL.
  • it can be more preferably 120 ⁇ g / mL or less.
  • it can be made to be the same as that of the particle
  • a sodium hydroxide aqueous solution was used as a titrant for titration of carboxyl groups and sulfonic acid groups. Dilute hydrochloric acid was used for the titration of amino groups.
  • the electrical conductivity was titrated under a nitrogen stream using a commercially available electrical conductivity meter (manufactured by Toa Decay Co., Ltd .: CM-60S).
  • the surface area of the particle was calculated from the formula of the surface area of the sphere using the catalog value of the particle diameter. It was confirmed that the catalog value of the particle diameter almost coincided with the value measured by a laser diffraction light scattering photometer (manufactured by Shimadzu Corporation: SALD-2300).
  • the reaction time (corresponding to the calcium re-coagulation time) when physiological saline was added instead of the reagent was defined as the blank time, and the value obtained by dividing the reaction time by the blank time was defined as the reaction rate index.
  • a smaller reaction rate index indicates that the coagulation reaction is promoted by the reagent.
  • the value obtained by dividing the final absorbance when the reagent was added by the final absorbance when physiological saline was added was defined as the absorbance change rate. The larger the absorbance change rate, the more the fibrin network grows. For all measurements, duplicate measurements were taken.
  • sample> The specimen was normal human plasma collected from a healthy person. Blood was collected using a vacuum blood collection tube (Becton Dickinson) containing a sodium citrate buffer and a 21 gauge blood collection needle. After blood collection, it was centrifuged at 3000 rpm for 10 minutes to separate plasma and blood cells.
  • a vacuum blood collection tube Becton Dickinson
  • Reagent 1 A 2.5 w / v% suspension of resin particles (Poly Science: 15913-10) having an average particle diameter of 0.05 ⁇ m made of a copolymer of polystyrene, acrylic acid and methacrylic acid was used. The particle concentration was 3.6 ⁇ 10 14 particles / mL. The surface density of the carboxyl group was 2.3 ⁇ 10 ⁇ mol / m 2 .
  • Reagent 2 The same procedure as in Reagent 1 was repeated except that resin particles having an average particle diameter of 0.1 ⁇ m made of a copolymer of polystyrene, acrylic acid and methacrylic acid (Poly Science: 16688-15) were used. Particle concentration was 4.6 ⁇ 10 13 cells / mL.
  • Reagent 3 The procedure was the same as that of Reagent 1 except that the resin particles were made of a copolymer of polystyrene, acrylic acid and methacrylic acid and had an average particle size of 0.5 ⁇ m (manufactured by Poly Science: 09836-15). The particle concentration was 3.6 ⁇ 10 11 particles / mL. The surface density of the carboxyl group was 1.5 ⁇ 10 ⁇ mol / m 2 .
  • Reagent 4 The procedure was the same as that of Reagent 1 except that resin particles made of a copolymer of polystyrene, acrylic acid and methacrylic acid and having an average particle diameter of 1.0 ⁇ m (manufactured by Poly Science: 08226-15) were used. The particle concentration was 4.6 ⁇ 10 10 particles / mL. The surface density of the carboxyl group was 8.0 ⁇ 10 ⁇ mol / m 2 .
  • mercerized cellulose is dispersed in 100 mL of water, 25 mg of TEMPO catalyst, 0.25 g of sodium bromide (NaBr) and 9.27% sodium hypochlorite (NaClO) aqueous solution as an oxidizing agent 10 mL And oxidation treatment was performed at room temperature and pH 10 for 2 hours. Thereafter, the reaction was stopped by adding a small amount of ethanol, and centrifuged at 10,000 g for 10 minutes to remove impurities, and then methanol was further added to the supernatant to precipitate COOH-CNF. The resulting precipitate was further centrifuged to recover COOH-CNF. The recovered COOH-CNF was dried at 80 ° C.
  • the average particle diameter of COOH-CNF in water was 5.4 ⁇ m.
  • the length of COOH-CNF was 2 ⁇ m to 5 ⁇ m and the width was 100 nm to 200 nm.
  • the length and width of COOH-CNF were the average of the lengths of 10 COOH-CNFs in the field of view observed with an electron microscope (manufactured by JEOL Ltd .: Jsm-6510, magnification 10,000).
  • the average particle diameter was measured with a laser diffraction light scattering photometer (manufactured by Shimadzu Corporation: SALD-2300).
  • the surface density of the carboxyl group was 1.2 ⁇ mol / m 2 .
  • Reagent 6 An aqueous suspension of sponge-like titanium oxide having an average particle size of 0.1 ⁇ m (manufactured by Erscreen Tohoku Co., Ltd .: PW116-3) was used.
  • the specific surface area of the sponge-like titanium oxide was 400 m 2 / g.
  • the particle concentration was 0.1 w / v%.
  • Reagent 7 Sponge-like titanium oxide used for reagent 6 was the same as reagent 5 except that it was heat-treated at 500 ° C. for 2 hours.
  • Reagent 8 The same procedure as in Reagent 1 except that resin particles having an amino group on the surface and an average particle size of 0.1 ⁇ m (Poly Science: 16586-5) was used. The particle concentration was 4.6 ⁇ 10 13 particles / mL. The surface density of the amino group was 14 ⁇ 10 ⁇ mol / m 2 .
  • Reagent 9 Reagent 1 was used except that resin particles having a sulfonic acid group on the surface and an average particle size of 0.5 ⁇ m (Poly Science Co., Ltd .: 19403-15) were used.
  • the surface density of the sulfonic acid group was 0.64 ⁇ 10 ⁇ mol / m 2 .
  • Reagent 10 The procedure was the same as that of Reagent 1 except that resin particles having a sulfonic acid group on the surface and having an average particle diameter of 1.0 ⁇ m (Poly Science, 19404-15) were used.
  • Reagent 11 The procedure was the same as that of Reagent 1 except that the resin particles were fixed with poly (2-methacryloyloxyethylphoshoryl choline) (PMPC).
  • the resin particles were resin particles having an amino group bonded through a linker having 3 carbon atoms and having an average particle size of 0.1 ⁇ m (manufactured by Poly Science: 16586-5).
  • the fixation of PMPC was performed via a succinimidyl group.
  • Reagent 12 It replaced with PMPC and was carried out similarly to the reagent 10 except having set it as the resin particle which fixed the block copolymer of MPC and acrylic acid shown in Formula 1. However, in Formula 1, m is 90 and n is 10 as preparation ratio.
  • Reagent 13 instead of PMPC, the same procedure as in Reagent 10 was performed except that resin particles fixed with block copolymers of MPC, acrylic acid and dimethylsiloxane shown in Formula 2 were used. However, in Formula 2, as the preparation ratio, (m + n) is 50, l is 50, m is 90, and n is 10.
  • Reagent 14 An aqueous suspension of cellulose nanofiber (Sugino Machine Co., Ltd .: BiNFi-s) was used. The particle concentration was 0.1 w / v%.
  • Reagent 15 A commercially available PT reagent (Sysmex Corporation: Thrombocheck PT) was used after dissolving in purified water according to the dosage.
  • Reagent 16 A commercially available APTT reagent (manufactured by SIEMENS: ACTIN) was used.
  • Table 1 shows the reaction rate index for each reagent.
  • the reaction rate indices at 100-fold dilution (250 ⁇ g / mL) for reagents 1 to 4 consisting of particles having carboxyl groups oriented on the surface were 0.59, 0.56, 0.49 and 0.60, respectively.
  • the reaction rate index at 100-fold dilution of the reagent 14 which is a commercially available PT reagent and the reagent 15 which is an APTT reagent was 0.55 and 0.49, respectively.
  • the reaction rate index of Reagents 1 to 4 is comparable to that of commercially available PT reagents and APTT reagents.
  • Reagents 1 to 4 promote the coagulation of plasma in the same manner as commercially available PT reagents and APTT reagents, and have the same blood coagulation promoting function as these reagents.
  • the reaction rate index was smaller than 1, and blood coagulation was promoted.
  • Reagent 5 consisting of COOH-CNF showed a reaction rate index of 0.88 at 10-fold dilution (100 ⁇ g / mL) and 0.74 at 100-fold dilution (10 ⁇ g / mL).
  • COOH-CNF also has a blood coagulation promoting function.
  • the reagent 6 composed of sponge-like titanium oxide showed a reaction rate index of 0.71 at 10-fold dilution (100 ⁇ g / mL) and 0.80 at 100-fold dilution (10 ⁇ g / mL).
  • the reaction rate index of the reagent 7 made of heat-treated sponge-like titanium oxide was 0.70 at 10-fold dilution and 0.81 at 100-fold dilution, indicating almost the same reaction rate index as that without heat treatment. It was.
  • the blood coagulation promoting function was recognized also about sponge-like titanium oxide particle.
  • Reagents 6 and 7 had a reaction rate index smaller than 1 even at 1000-fold dilution (1 ⁇ g / mL), and promoted blood coagulation.
  • the reaction rate index was 0.94 at a 100-fold dilution of Reagent 8 consisting of particles having amino groups on the surface, and a slight promotion of blood coagulation was observed.
  • the reaction rate index was larger than 1, and the blood coagulation accelerating function was not observed when the surface did not contain particles having carboxyl groups oriented or sponge-like titanium oxide particles.
  • Table 2 shows the absorbance change rate for each reagent.
  • Reagent 1 to Reagent 8 having a reaction rate index of less than 1 Reagent 1 consisting of particles having an average particle diameter of 0.05 ⁇ m had an absorbance change rate of 1 or less at 100-fold dilution.
  • Reagent 3 composed of particles having an average particle diameter of 0.5 ⁇ m and Reagent 4 composed of particles having an average particle diameter of 1.0 ⁇ m also had a slightly smaller absorbance change rate at 100-fold dilution than other reagents.
  • Reagent 9 composed of particles having an average particle diameter of 0.5 ⁇ m and Reagent 10 composed of particles having an average particle diameter of 1.0 ⁇ m are other reagents.
  • the rate of change in absorbance at 100-fold dilution was slightly smaller than that of.
  • FIGS. 2 to 6 show SEM photographs of the fibrin network obtained when the reagents 1, 3, 6, and 11 were diluted 100 times and in the blank, respectively.
  • fibrin fibers are finer than other reagents and blanks.
  • reagent 1 also has a sufficient blood coagulation promoting function and is useful as a blood coagulation promoter.
  • the blood coagulation promoter of the present disclosure exhibits a stable blood coagulation function and is useful as a blood coagulation function test agent or the like.

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Abstract

　Cet accélérateur de coagulation du sang et cet agent pour tester la fonction de caillots sanguins comprennent des particules comportant des groupes carboxyle orientés sur leur surface, ou des particules d'oxyde de titane de type éponge.
PCT/JP2015/003003 2014-06-18 2015-06-16 Accélérateur de coagulation du sang, et agent pour tester la fonction de caillots sanguins à l'aide de celui-ci WO2015194165A1 (fr)

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Cited By (5)

* Cited by examiner, † Cited by third party
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WO2018012629A1 (fr) * 2016-07-14 2018-01-18 東洋製罐グループホールディングス株式会社 Fibres de cellulose comprenant des nanofibres de cellulose, et procédé de fabrication de celles-ci
JP7444572B2 (ja) 2019-09-24 2024-03-06 シスメックス株式会社 活性化部分トロンボプラスチン時間測定用試薬及び試薬キット、並びにエラグ酸化合物の沈殿を抑制する方法
WO2024080372A1 (fr) * 2022-10-14 2024-04-18 積水メディカル株式会社 Agent modifiant le temps de coagulation sanguine d'un échantillon de sang défaillant en facteur de coagulation ｖｉｉｉ, ｉｘ ou ｘｉ, et réactif pour mesure du temps de thromboplastine partielle activée
WO2024080371A1 (fr) * 2022-10-14 2024-04-18 積水メディカル株式会社 Agent raccourcissant le temps de coagulation sanguine d'un échantillon de sang défaillant en facteur de coagulation ｘｉｉ
WO2024080373A1 (fr) * 2022-10-14 2024-04-18 積水メディカル株式会社 Réactif pour mesure du temps de thromboplastine partielle activée, et agent modifiant le temps de coagulation sanguine d'un échantillon de sang positif à l'anticoagulant lupique ou d'un échantillon de sang à teneur en héparine

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08154697A (ja) * 1994-12-08 1996-06-18 Sekisui Chem Co Ltd 血液検査用容器
JP2006200997A (ja) * 2005-01-19 2006-08-03 Sekisui Chem Co Ltd 血液凝固促進剤
WO2012156216A1 (fr) * 2011-05-16 2012-11-22 The Swatch Group Research And Development Ltd Augmentation de la durée de vie de stockage d'un capteur de thrombine

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08154697A (ja) * 1994-12-08 1996-06-18 Sekisui Chem Co Ltd 血液検査用容器
JP2006200997A (ja) * 2005-01-19 2006-08-03 Sekisui Chem Co Ltd 血液凝固促進剤
WO2012156216A1 (fr) * 2011-05-16 2012-11-22 The Swatch Group Research And Development Ltd Augmentation de la durée de vie de stockage d'un capteur de thrombine

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
KOZUE OKANO ET AL.: "Kobunshi Gosei Kagobutsu o Mochiita Gyoko Sokutei Shiyaku no Kaihatsu", JOURNAL OF THE JAPANESE SOCIETY FOR LABORATORY HEMATOLOGY, vol. 15, 20 June 2014 (2014-06-20), pages S160 *
SOMNATH C. ROY ET AL.: "The effect of Ti02 nanotubes in the enhancement of blood clotting for the control of hemorrhage", BIOMATERIALS, vol. 28, no. Issues 31, November 2007 (2007-11-01), pages 4667 - 4672, XP022207007, ISSN: 0142-9612 *
WEI-EN YANG ET AL.: "Nano/submicron-scale TiO2 network on titanium surface for dental implant application", JOURNAL OF ALLOYS AND COMPOUNDS, vol. 479, no. Issues 1-2, 24 June 2009 (2009-06-24), pages 642 - 647, XP026108891, ISSN: 0925-8388 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018012629A1 (fr) * 2016-07-14 2018-01-18 東洋製罐グループホールディングス株式会社 Fibres de cellulose comprenant des nanofibres de cellulose, et procédé de fabrication de celles-ci
JP7444572B2 (ja) 2019-09-24 2024-03-06 シスメックス株式会社 活性化部分トロンボプラスチン時間測定用試薬及び試薬キット、並びにエラグ酸化合物の沈殿を抑制する方法
WO2024080372A1 (fr) * 2022-10-14 2024-04-18 積水メディカル株式会社 Agent modifiant le temps de coagulation sanguine d'un échantillon de sang défaillant en facteur de coagulation ｖｉｉｉ, ｉｘ ou ｘｉ, et réactif pour mesure du temps de thromboplastine partielle activée
WO2024080371A1 (fr) * 2022-10-14 2024-04-18 積水メディカル株式会社 Agent raccourcissant le temps de coagulation sanguine d'un échantillon de sang défaillant en facteur de coagulation ｘｉｉ
WO2024080373A1 (fr) * 2022-10-14 2024-04-18 積水メディカル株式会社 Réactif pour mesure du temps de thromboplastine partielle activée, et agent modifiant le temps de coagulation sanguine d'un échantillon de sang positif à l'anticoagulant lupique ou d'un échantillon de sang à teneur en héparine
JP7473282B1 (ja) 2022-10-14 2024-04-23 積水メディカル株式会社 凝固第ｘｉｉ因子欠乏血液検体の血液凝固時間短縮剤
JP7473283B1 (ja) 2022-10-14 2024-04-23 積水メディカル株式会社 活性化部分トロンボプラスチン時間測定のための試薬、及びループスアンチコアグラント陽性血液検体又はヘパリン含有血液検体の血液凝固時間調節剤
JP7473281B1 (ja) 2022-10-14 2024-04-23 積水メディカル株式会社 凝固第ｖｉｉｉ、ｉｘ又はｘｉ因子欠乏血液検体の血液凝固時間調節剤、及び活性化部分トロンボプラスチン時間測定のための試薬

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