US20120064553A1 - Blood coagulation time prolonging agent - Google Patents

Blood coagulation time prolonging agent Download PDF

Info

Publication number
US20120064553A1
US20120064553A1 US13/321,732 US201013321732A US2012064553A1 US 20120064553 A1 US20120064553 A1 US 20120064553A1 US 201013321732 A US201013321732 A US 201013321732A US 2012064553 A1 US2012064553 A1 US 2012064553A1
Authority
US
United States
Prior art keywords
blood coagulation
coagulation time
blood
measuring
alkyl group
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US13/321,732
Other languages
English (en)
Inventor
Chizuru Morikawa
Remi Nakamura
Mitsuaki Yamamoto
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sekisui Medical Co Ltd
Original Assignee
Sekisui Medical Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Sekisui Medical Co Ltd filed Critical Sekisui Medical Co Ltd
Assigned to SEKISUI MEDICAL CO., LTD. reassignment SEKISUI MEDICAL CO., LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: NAKAMURA, REMI, MORIKAWA, CHIZURU, YAMAMOTO, MITSUAKI
Publication of US20120064553A1 publication Critical patent/US20120064553A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C279/00—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
    • C07C279/02—Guanidine; Salts, complexes or addition compounds thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C279/00—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
    • C07C279/04—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of guanidine groups bound to acyclic carbon atoms of a carbon skeleton
    • C07C279/14—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of guanidine groups bound to acyclic carbon atoms of a carbon skeleton being further substituted by carboxyl groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C281/00—Derivatives of carbonic acid containing functional groups covered by groups C07C269/00 - C07C279/00 in which at least one nitrogen atom of these functional groups is further bound to another nitrogen atom not being part of a nitro or nitroso group
    • C07C281/16—Compounds containing any of the groups, e.g. aminoguanidine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/56—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/745—Assays involving non-enzymic blood coagulation factors

Definitions

  • the present invention relates to a reagent for measuring blood coagulability typified by a reagent for measuring fibrinogen; specifically relates to a blood coagulation time prolonging agent used in a blood coagulation activator-containing reagent and/or in a specimen diluent for measuring blood coagulability, and to a reagent for measuring blood coagulability using this blood coagulation time prolonging agent.
  • a blood coagulation mechanism is broadly divided into two pathways in general. One is an intrinsic pathway, which is initiated by contact activation of blood coagulation factor XII with a foreign substance, and finally produces thrombin through multistep reactions, and the other is an extrinsic pathway, which is initiated by activation of blood coagulation factor X with blood coagulation factor VII and tissue thromboplastin, and produces thrombin in the same manner as described above ( FIG. 1 ). In both of the pathways, coagulation eventually occurs by conversion of fibrinogen to fibrin through an action of produced thrombin. In order to clarify the presence or absence of abnormality or a cause of the abnormality in such blood coagulation mechanism, some blood coagulability tests using a blood coagulation activator are available and widely used in actual clinical examinations.
  • the blood coagulation activator and the blood coagulability tests using the same are as follows:
  • prothrombin time measurement an activity measurement of factor II, V, VII, or X using the prothrombin time measurement, and a measurement of complex factors (a thrombo test, a hepaplastin test, and the like);
  • a partial thromboplastin time measurement an activated partial thromboplastin time measurement, an activity measurement of factor VIII, IX, XI, or XII, prekallikrein, or high molecular kininogen using the activated partial thromboplastin time measurement, a viper venom time measurement, a quantitative determination of factor X using the viper venom time measurement, a lupus anti-coagulant (LA) measurement using a diluted viper venom time measurement, a protein C activity measurement, and a thromboplastin generation test.
  • a viper venom time measurement a quantitative determination of factor X using the viper venom time measurement
  • LA lupus anti-coagulant
  • a time from initiation of coagulation by mixing a reagent including a blood coagulation activator and the like with a specimen from a patient to final conversion of fibrinogen into fibrin to be deposited is measured.
  • the mechanical detection method refers to a method including: monitoring by a magnetic force or the like a magnetic substance or the like loaded into a reaction solution; and detecting a decrease in mobility of the magnetic substance due to an increase in viscosity by coagulation.
  • the optical detection method refers to a method including detecting white turbidity of a reaction solution due to coagulation as a change in transmitted light or scattered light, and is most widely used because the method is relatively simple. These are generally detected by an automated analyzer at present.
  • FIG. 2 One example of a change curve for scattered light intensity obtained in a scattered light detection method is shown in FIG. 2 .
  • the point A indicates a point at which the coagulation is initiated by mixing a reagent including a blood coagulation activator, subsequently fibrin deposition is initiated through multistep reactions, and as a result, a change in scattered light intensity appears after the point B. Fibrinogen is in turn consumed and almost depleted in the reaction solution. Then, the scattered light intensity exhibits no change, the curve becomes flat as shown at the point C, and the coagulation is terminated.
  • a coagulation time is calculated with known calculation parameters based on such change curve for the scattered light intensity (Patent document 1).
  • the reagent for measuring blood coagulability desirably has a property in which an optical change is largely displayed.
  • a substance for prolonging the coagulation time is generally added to adjust the coagulation time to a desired time.
  • a blood coagulation time prolonging agent is exemplified by halide salts of alkali metals and alkali earth metals, including sodium chloride in Patent document 2, and by sodium propionate in Patent document 3.
  • halide salts of alkali metals and alkali earth metals including sodium chloride in Patent document 2, and by sodium propionate in Patent document 3.
  • Patent documents 4 and 5 For the purpose of solving the problems in those conventional methods, there is proposed a method including using a reagent supplemented with a high molecular material such as polyethylene glycol, polyvinyl alcohol or high molecular polysaccharide.
  • the inventors of the present invention have searched a blood coagulation time prolonging agent which enhances an optical change using various compounds, and have found that a guanidine compound having a specific structure or a salt thereof has not only an action of prolonging a blood coagulation time but also an action of enhancing an optical change upon coagulation, and the use thereof as an additive for a reagent for measuring blood coagulability allows the blood coagulability to be measured correctly with high sensitivity, and have completed the present invention.
  • the present invention provides a blood coagulation time prolonging agent, including, as an active ingredient, a guanidine compound represented by the following formula (1) or an acid addition salt thereof:
  • R 1 represents a hydrogen atom, an amino group, or an alkyl group which may have a substituent.
  • the present invention provides a blood coagulation activator-containing reagent or a specimen diluent for measuring blood coagulability, including the above-mentioned blood coagulation time prolonging agent.
  • the present invention provides a blood coagulation activator-containing reagent for measuring fibrinogen including thrombin as the blood coagulation activator or a specimen diluent for measuring fibrinogen, containing the above-mentioned blood coagulation time prolonging agent.
  • the present invention provides a use of a guanidine compound represented by the following formula (1) or an acid addition salt thereof for prolonging a blood coagulation time:
  • R 1 represents a hydrogen atom, an amino group, or an alkyl group which may have a substituent.
  • the above-mentioned guanidine compound or acid addition salt thereof is used in combination with the blood coagulation activator. In another embodiment, the above-mentioned guanidine compound or acid addition salt thereof is used in combination with the specimen diluent for measuring blood coagulability.
  • the present invention provides a method of prolonging a blood coagulation time, including adding to a plasma specimen a guanidine compound represented by the following formula (1) or an acid addition salt thereof:
  • R 1 represents a hydrogen atom, an amino group, or an alkyl group which may have a substituent.
  • the addition of the above-mentioned guanidine compound or acid addition salt thereof is carried out in combination with the specimen diluent for measuring blood coagulability. In another embodiment, the addition of the above-mentioned guanidine compound or acid addition salt thereof is carried out in combination with the blood coagulation activator.
  • the present invention provides a method of measuring blood coagulability, including the steps of: preparing a reaction solution containing a plasma specimen, a blood coagulation activator, and the above-mentioned guanidine compound or acid addition salt thereof; and measuring a coagulation time of the reaction solution.
  • the present invention provides a method of determining a fibrinogen concentration in the plasma specimen by comparing a measured value of the coagulation time using the plasma specimen with a measured value obtained by similarly measuring the coagulation time using a diluted standard solution in place of the plasma specimen.
  • fibrinogen level, PT, and APTT for blood coagulability can be measured correctively because the optical change is enhanced.
  • FIG. 1 is a view schematically showing a blood coagulation mechanism in a clinical examination.
  • FIG. 2 is a graph showing one example of a change curve for scattered light intensity obtained by a scattered light detection method when blood coagulability is measured.
  • FIG. 3 is a graph showing a relationship between an addition amount and a coagulation time of each compound.
  • FIG. 4 is a graph showing a relationship between a coagulation time and a maximum of a change in scattered light intensity when the coagulation time is controlled by the addition of each compound.
  • FIG. 5 is a graph showing change curves for scattered light intensity obtained by using reagents according to conventional methods and a reagent of the present invention.
  • a blood coagulation time prolonging agent of the present invention contains a compound represented by the formula (1) or a salt thereof as an active ingredient.
  • R 1 represents a hydrogen atom, an amino group, or an alkyl group which may have a substituent.
  • the alkyl group is preferably an alkyl group having 1 to 6 carbon atoms, particularly preferably an alkyl group having 2 to 5 carbon atoms.
  • a group which may serve as a substituent of the alkyl group is exemplified by a carboxy group, an alkoxycarbonyl group, and an amino group.
  • the alkyl group is preferably substituted with both of the carboxy group and the amino group or with both of the alkoxycarbonyl group and the amino group.
  • the compound represented by the formula (1) examples include guanidine, aminoguanidine, arginine, and an arginine alkyl ester. Of those, aminoguanidine is particularly preferred because an action of enhancing an optical change is large and the correct measurement can be performed.
  • the acid addition salt of the compound of the formula (1) include a hydrochloride, a sulfate, a nitrate, a phosphate, and a sulfamate.
  • the blood coagulation time prolonging agent of the present invention has not only an action of prolonging a blood coagulation time but also an action of enhancing a turbidity change of a reaction solution in the measurement of blood coagulability. This indicates that a network of fibrin, which is a final product of blood coagulation, is more firmly formed. Therefore, the blood coagulation time prolonging agent of the present invention can be used in, for example, an optical detection method including directly detecting a turbidity change and a mechanical detection method including monitoring a formation state of a network of fibrin with a magnetic substance or the like. Of those, it is preferred to use the blood coagulation time prolonging agent in the optical detection method.
  • a method of detecting a scattered light amount and a method of detecting a transmitted light amount are available as the optical detection method. Of those, it is preferred to use the blood coagulation time prolonging agent in the method of detecting a scattered light amount.
  • a blood coagulability test to which the blood coagulation time prolonging agent of the present invention can be applied may be any test including measuring a time from the initiation of coagulation by mixing a reagent containing a blood coagulation activator and the like with a patient specimen to the final conversion of fibrinogen into fibrin to be deposited.
  • Examples thereof include a fibrinogen measurement, an ATIII measurement, a thrombin time measurement, a prothrombin time measurement, an activity measurement of factor II, V, VII, or X using a prothrombin time, a measurement of complex factors (a thrombo test, a hepaplastin test, and the like), a partial thromboplastin time measurement, an activated partial thromboplastin time measurement, an activity measurement of factor VIII, IX, XI, or XII, prekallikrein, or high molecular kininogen using the activated partial thromboplast in time measurement, a viper venom time measurement, a quantitative determination of factor X using the viper venom time measurement, a lupus anti-coagulant (LA) measurement using a diluted viper venom time measurement, a protein C activity measurement, a thromboplastin generation test, and any other detection of abnormal coagulation.
  • a fibrinogen measurement an ATIII measurement, a thrombin
  • a time from the addition of a blood coagulation activator-containing reagent e.g., a thrombin-containing reagent in the fibrinogen measurement
  • the specimen is appropriately diluted with a specimen diluent if necessary.
  • the blood coagulation time prolonging agent of the present invention has only to be contained in a measurement system.
  • the blood coagulation time prolonging agent may be contained in the blood coagulation activator-containing reagent, or may be contained in the specimen diluent. That is, for example, in the fibrinogen measurement, the blood coagulation time prolonging agent of the present invention may be contained in the thrombin-containing reagent, or may be contained in the specimen diluent.
  • the above-mentioned blood coagulation activator-containing reagent has only to be one used for the above-mentioned blood coagulability test, and examples thereof include a thrombin-containing reagent, a tissue thromboplastin-containing reagent, and a phospholipid-containing reagent.
  • the above-mentioned specimen diluent has only to be one used for the above-mentioned blood coagulability test, and examples thereof include Good's buffers such as MES, Bis-Tris, ADA, PIPES, ACES, MOPSO, BES, MOPS, TES, HEPES, DIPSO, TAPSO, POPSO, HEPPSO, EPPS, Tricine, Bicine, TAPS, and CHES, a citrate buffer, aphosphatebuffer, an acetate buffer, an imidazole buffer, a barbital buffer, saline, and water.
  • Good's buffers such as MES, Bis-Tris, ADA, PIPES, ACES, MOPSO, BES, MOPS, TES, HEPES, DIPSO, TAPSO, POPSO, HEPPSO, EPPS, Tricine, Bicine, TAPS, and CHES, a citrate buffer, aphosphatebuffer, an acetate buffer, an imidazo
  • the blood coagulation time prolonging agent of the present invention is preferably contained in the reaction solution so that a coagulation time which is measurable with high accuracy is obtained in the blood coagulability test.
  • the blood coagulation time prolonging agent is preferably contained in the reaction solution so that the coagulation time is 5 to 50 seconds, preferably 7 to 20 seconds, more preferably 9 to 15 seconds when normal plasma (concentration: about 300 mg/dL) is measured.
  • the concentration of aminoguanidine in the reaction solution for the fibrinogen measurement i.e., the concentration of aminoguanidine in a mixed solution of a specimen, a specimen diluent, and a thrombin-containing reagent is 10 to 500 mM, preferably 20 to 200 mM, more preferably 30 to 120 mM.
  • the concentration of aminoguanidine in the specimen diluent is 10 to 900 mM, preferably 30 to 400 mM, more preferably 50 to 200 mM.
  • the blood coagulation time prolonging agent is preferably contained in the reaction solution so that the coagulation time is 9 to 60 seconds, preferably 9 to 30 seconds, more preferably 10 to 16 seconds when normal plasma (activity: about 100%) is measured.
  • the blood coagulation time prolonging agent is preferably contained in the reaction solution so that the coagulation time is 15 to 80 seconds, preferably 15 to 60 seconds, more preferably 20 to 50 seconds when normal plasma is measured.
  • the blood coagulation time prolonging agent has to be contained in the reaction solution so that a coagulation time suitable for a measurement method is obtained.
  • known high molecular polysaccharides and synthetic polymers can be contained in the reagent for measuring blood coagulability including the specimen diluent and the blood coagulation activator-containing reagent used for the blood coagulability test.
  • high molecular polysaccharides include dextran such as dextran 40, dextran 70, dextran 200000, or dextran 500000, and Ficoll, and the like.
  • dextran such as dextran 40, dextran 70, dextran 200000, or dextran 500000, and Ficoll, and the like.
  • One kind of those high molecular polysaccharides may be used alone, or two or more kinds thereof may be used in combination.
  • Those high molecular polysaccharides are each added in an amount so that the concentration in the reagent for measuring blood coagulability is suitably 0.01 to 10 W/V %, more preferably 0.1 to 5 W/V %.
  • the synthetic polymers include: polyvinyl alcohol such as polyvinyl alcohol 500, polyvinyl alcohol 1500, or polyvinyl alcohol 2000; polyethylene glycol such as polyethylene glycol 1500, polyethylene glycol 2000, polyethylene glycol 4000, polyethylene glycol 6000, polyethylene glycol 8000, or polyethylene glycol 20000; and polyvinylpyrrolidone.
  • polyvinyl alcohol such as polyvinyl alcohol 500, polyvinyl alcohol 1500, or polyvinyl alcohol 2000
  • polyethylene glycol such as polyethylene glycol 1500, polyethylene glycol 2000, polyethylene glycol 4000, polyethylene glycol 6000, polyethylene glycol 8000, or polyethylene glycol 20000
  • polyvinylpyrrolidone One kind of those synthetic polymers may be used alone, or two or more kinds thereof may be used in combination.
  • the amount of each of those synthetic polymers to be added is not particularly limited, and the concentration thereof in the reagent for measuring blood coagulability is preferably 0.01 to 10W/V %, more
  • the reagent for measuring blood coagulability can also contain a buffer, a calcium ion, an antagonist of an anticoagulant, and the like in addition to the blood coagulation time prolonging agent of the present invention.
  • the buffer As a specific example of the buffer, a buffer having a buffering ability in the range of pH 4 to 9 is appropriately selected and used.
  • the buffer can be selected from Good's buffers such as MES, Bis-Tris, ADA, PIPES, ACES, MOPSO, BES, MOPS, TES, HEPES, DIPSO, TAPSO, POPSO, HEPPSO, EPPS, Tricine, Bicine, TAPS, and CHES, citric acid, phosphoric acid, acetic acid, imidazole, barbital, GTA, and the like, and one kind thereof maybe used alone, or two or more or kinds thereof may be used in combination.
  • the amount of the buffer to be added is not particularly limited as long as the buffer has a buffering ability, and the concentration in the reagent for measuring blood coagulability is preferably 1 to 1,000 mM, more preferably 5 to 500 mM.
  • Water-soluble calcium compounds such as calcium chloride, calcium lactate, calciumgluconate, calcium glucuronate, and calcium tartrate are used as the calcium ion.
  • One kind of those calcium compounds may be used alone, or two or more kinds thereof may be used in combination.
  • the amount of each of the calcium compounds to be used is not particularly limited as long as the calcium compounds can help a coagulation reaction, and the concentration thereof in the reagent for measuring blood coagulability is preferably 5 mM to 100 mM, and more preferably 10 mM to 50 mM.
  • Protamine, polybrene (hexadimethrine bromide), and the like are used as the antagonist of the anticoagulant.
  • One kind of those antagonists of the anticoagulant may be used alone, or two or more kinds thereof may be used in combination.
  • the amount of the antagonist of the anticoagulant to be used is not particularly limited as long as the antagonist can sufficiently neutralize an anticoagulant such as heparin contained in specimen plasma, and the concentration in the reagent for measuring blood coagulability is preferably 10 ⁇ 5 to 10 ⁇ 2 W/V %, more preferably 5 ⁇ 10 ⁇ 4 to 5 ⁇ 10 ⁇ 3 W/V %.
  • the reagent for measuring blood coagulability may be a liquid product, a frozen product, or a dried product, and the dried product is dissolved by adding purified water or a buffer.
  • an appropriate preservative may be added to the reagent for measuring blood coagulability.
  • the preservative can be selected from ciprofloxacin, propionic acid, sodium benzoate, sodium azide, ProClin 300, and the like, and one kind thereof may be used alone, or two or more kinds thereof may be used in combination. Further, salts such as sodium chloride, common stabilizers such as amino acids and sugars, and the like may be contained as needed.
  • the concentration described above is the concentration in the solution, and the concentration of the dried product or the like is the concentration when dissolved in water, a buffer, or the like before use.
  • the measurement of blood coagulability of the present invention can be conducted with a usual method. Taking the fibrinogen measurement as an example, a standard solution is diluted with a specimen diluent 5-fold, 10-fold, and 20-fold (a dilution rate can be adjusted appropriately). Next, two volumes of each diluted standard solution is warmed at 37° C. for 3 minutes, one volume of a thrombin reagent previously warmed at 37° C. is added, and a coagulation time is measured. Subsequently, a measured value of the each diluted standard solution is plotted on a graph. A plasma specimen is diluted with a specimen diluent 10-fold (a dilution rate can be adjusted appropriately), the diluted specimen is measured likewise, and the concentration can be obtained from the graph based on the obtained coagulation time.
  • thrombin time method When a fibrinogen concentration in a plasma specimen is measured by a thrombin time method, in general, thrombin is added to plasma to measure a coagulation time, and the concentration is determined from a standard curve prepared using standard solutions having known fibrinogen concentrations.
  • 90 ⁇ L of a specimen diluent are added to and mixed with 10 ⁇ L of specimen plasma, subsequently 50 ⁇ L of a thrombin-containing reagent are added as a blood coagulation activator-containing reagent, and a coagulation time is measured.
  • a maximum of a change in scattered light intensity was calculated as ⁇ H, and its relationship with the coagulation time was shown in FIG. 4 .
  • ⁇ H was remarkably decreased along with the prolonged coagulation time in the conventional methods using sodium chloride and sodium bromide, while large ⁇ H was kept in the present invention.
  • the change in scattered light intensity is larger in the present invention than in the conventional methods, and it is found that the coagulation time can be measured more correctly in the present invention.
  • Example 1 Commercially available normal plasma was measured in the same manner as in Example 1 using the specimen diluent (HEPES buffer, pH 7.5) in which the amount of the blood coagulation time prolonging agent added was adjusted so as to obtain the similar coagulation time.
  • the comparative results of the maximum of a change ⁇ H in scattered light intensity and of the simultaneous reproducibility of the measurements between in the conventional method and in the present invention were shown in Table 1.
  • the blood coagulation time prolonging agent of the present invention i.e., arginine hydrochloride, arginine methyl ester, guanidine hydrochloride, guanidine phosphate, guanidine sulfamate, or aminoguanidine hydrochloride drastically enhances ⁇ H and further enhances the accuracy of the measurement (simultaneous reproducibility or the like). It was also found that aminoguanidine hydrochloride had a particularly excellent effect of improving the accuracy of the measurement (simultaneous reproducibility or the like).
  • a specimen diluent containing sodium chloride as the blood coagulation time prolonging agent was defined as a conventional method A and a specimen diluent further containing polyethylene glycol (molecular weight: 20000) (hereinafter, referred to as PEG 20000) as an enhancer of a change in light intensity in the conventional method A was defined as a conventional method B, and the specimen diluents were compared with a specimen diluent employing aminoguanidine hydrochloride (present invention) which served as both of the blood coagulation time prolonging agent and the enhancer of a change in light intensity.
  • an HEPES buffer pH 7.5
  • the concentration of PEG 20000 to be added in the conventional method B was set to 0.1%, because a large amount of PEG 20000 could increase the viscosity of diluent and reduce the accuracy of sampling.
  • the coagulation time of commercially available normal plasma was measured in the same manner as in Example 1 using each specimen diluent. As a result, a change curve for scattered light intensity as shown in FIG. 5 was obtained. Further, the coagulation time and ⁇ H were shown in Table 2.
  • the change of the scattered light intensity in the conventional method B with the addition of PEG 20000 became larger than in the conventional method A without the addition. However, it is found that, by employing the present invention, the change of the scattered light intensity is more drastically enhanced and the coagulation time can be measured more correctly.
  • a method B invention Coagulation time NaCl 150 mM NaCl 150 mM Aminoguanidine prolonging agent hydrochloride Enhancer of change — PEG 90 mM in light intensity 20000 0.1% Coagulation time 11.6 seconds 10.4 seconds 10.4 seconds Change in light 2,194 2,244 3,444 intensity ⁇ H Mean of three times measurement
US13/321,732 2009-05-20 2010-05-20 Blood coagulation time prolonging agent Abandoned US20120064553A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2009121834 2009-05-20
PCT/JP2010/003401 WO2010134345A1 (ja) 2009-05-20 2010-05-20 血液凝固時間延長剤

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2010/003401 A-371-Of-International WO2010134345A1 (ja) 2009-05-20 2010-05-20 血液凝固時間延長剤

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US14/153,739 Division US20140127727A1 (en) 2009-05-20 2014-01-13 Blood coagulation time prolonging agent

Publications (1)

Publication Number Publication Date
US20120064553A1 true US20120064553A1 (en) 2012-03-15

Family

ID=43126036

Family Applications (3)

Application Number Title Priority Date Filing Date
US13/321,732 Abandoned US20120064553A1 (en) 2009-05-20 2010-05-20 Blood coagulation time prolonging agent
US14/153,739 Abandoned US20140127727A1 (en) 2009-05-20 2014-01-13 Blood coagulation time prolonging agent
US14/658,911 Active US9733262B2 (en) 2009-05-20 2015-03-16 Method for measuring blood coagulation

Family Applications After (2)

Application Number Title Priority Date Filing Date
US14/153,739 Abandoned US20140127727A1 (en) 2009-05-20 2014-01-13 Blood coagulation time prolonging agent
US14/658,911 Active US9733262B2 (en) 2009-05-20 2015-03-16 Method for measuring blood coagulation

Country Status (7)

Country Link
US (3) US20120064553A1 (ko)
EP (2) EP3018481B1 (ko)
JP (2) JPWO2010134345A1 (ko)
KR (2) KR101882408B1 (ko)
CN (1) CN102428372A (ko)
CA (1) CA2762611C (ko)
WO (1) WO2010134345A1 (ko)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5903215B2 (ja) * 2011-02-28 2016-04-13 シスメックス株式会社 ループスアンチコアグラント検出用試薬キット及びループスアンチコアグラントの存否を判定する方法
CN110559435B (zh) 2013-09-11 2023-10-20 伊戈尔生物药品股份有限公司 包含离子性液体的液体蛋白质制剂
US11471479B2 (en) 2014-10-01 2022-10-18 Eagle Biologics, Inc. Polysaccharide and nucleic acid formulations containing viscosity-lowering agents
CN105372424A (zh) * 2015-11-12 2016-03-02 武汉景川诊断技术股份有限公司 肝促凝血活酶试验测定试剂盒及其制备方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5851836A (en) * 1994-09-02 1998-12-22 Nippon Shoji Kaisha Ltd. Method for determining fibrinogen and reagent for determination thereof
US6037356A (en) * 1997-11-26 2000-03-14 3-Dimensional Pharmaceuticals, Inc. Heteroaryl aminoguanidines and alkoxyguanidines and their use as protease inhibitors
US20040077092A1 (en) * 2002-05-16 2004-04-22 Hans Wielinger Control and calibration material for blood coagulation tests
US7741124B2 (en) * 2003-06-04 2010-06-22 Stief Thomas W Detection procedures for fibrinogen and/or fibrinogen derivatives

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3180824B2 (ja) 1991-08-30 2001-06-25 シスメックス株式会社 血液凝固試薬
JP3074611B2 (ja) 1991-08-30 2000-08-07 シスメックス株式会社 血液凝固測定方法
US5358853A (en) 1992-08-03 1994-10-25 Akzo Av Liquid thromboplastin reagent
JPH0815263A (ja) 1994-06-24 1996-01-19 Shimadzu Corp 血液凝固分析装置
DE19549118C2 (de) * 1995-12-29 2000-07-13 Thomas W Stief Hämostaseaktivierungs-Inhibitor und Verfahren zum Hemmen der Hämostaseaktivierung in Blut oder anderen biologischen Flüssigkeiten
TW499412B (en) * 1996-11-26 2002-08-21 Dimensional Pharm Inc Aminoguanidines and alkoxyguanidines as protease inhibitors
CN1504580A (zh) * 2002-11-28 2004-06-16 希森美康株式会社 凝血酶试剂以及检查试剂套装
JP2004191367A (ja) * 2002-11-28 2004-07-08 Sysmex Corp トロンビン試薬及び検査試薬キット
EP1672369A1 (de) * 2004-12-16 2006-06-21 Roche Diagnostics GmbH Verfahren zur Bestimmung von Blutkoagulationsparametern, insbesondere in Durchflussmesssystemen

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5851836A (en) * 1994-09-02 1998-12-22 Nippon Shoji Kaisha Ltd. Method for determining fibrinogen and reagent for determination thereof
US6037356A (en) * 1997-11-26 2000-03-14 3-Dimensional Pharmaceuticals, Inc. Heteroaryl aminoguanidines and alkoxyguanidines and their use as protease inhibitors
US20040077092A1 (en) * 2002-05-16 2004-04-22 Hans Wielinger Control and calibration material for blood coagulation tests
US7741124B2 (en) * 2003-06-04 2010-06-22 Stief Thomas W Detection procedures for fibrinogen and/or fibrinogen derivatives

Also Published As

Publication number Publication date
EP2434290B1 (en) 2016-02-10
EP3018481A1 (en) 2016-05-11
EP3018481B1 (en) 2018-02-21
KR20180019781A (ko) 2018-02-26
JP5840273B2 (ja) 2016-01-06
US9733262B2 (en) 2017-08-15
KR20120024610A (ko) 2012-03-14
CA2762611C (en) 2017-03-28
EP2434290A1 (en) 2012-03-28
WO2010134345A1 (ja) 2010-11-25
JPWO2010134345A1 (ja) 2012-11-08
CN102428372A (zh) 2012-04-25
EP2434290A4 (en) 2013-01-30
CA2762611A1 (en) 2010-11-25
US20140127727A1 (en) 2014-05-08
JP2015042652A (ja) 2015-03-05
US20150185236A1 (en) 2015-07-02
KR101882408B1 (ko) 2018-07-27

Similar Documents

Publication Publication Date Title
Kachhawa et al. Study of the stability of various biochemical analytes in samples stored at different predefined storage conditions at an accredited laboratory of India
US9733262B2 (en) Method for measuring blood coagulation
US7732210B2 (en) Multi-analyte reference solutions
WO2002079375A1 (en) Rapid assessment of coagulation activity in whole blood
JP6025105B2 (ja) ループスアンチコアグラント検出用血液凝固時間の測定方法
CN104076156A (zh) 活化部分凝血活酶时间测定法和活化部分凝血活酶时间测定用试剂盒
JP2011133396A (ja) 活性化部分トロンボプラスチン時間測定試薬、活性化部分トロンボプラスチン時間測定方法、及び血液凝固抑制物質の有無の判定方法
Handeland et al. Simplified assay for antithrombin III activity using chromogenic peptide substrate: Manual and automated method
WO1994007145A1 (fr) Procede de determination de l'activite du facteur de coagulation sanguine xiii et trousse de reactifs appropries
JP2008145278A (ja) ガストリン放出ペプチド前駆体の免疫学的測定方法
US20220120768A1 (en) Reagent for measuring fibrinogen
Inada et al. Faster determination of clottable fibrinogen in human plasma: an improved method and kinetic study.
US5614414A (en) Liquid lipoprotein control
JP6058726B2 (ja) 血液凝固反応における血液凝固時間延長剤
CN110760565A (zh) 一种对氧磷酶1的检测试剂盒及制备方法
JP7473282B1 (ja) 凝固第ｘｉｉ因子欠乏血液検体の血液凝固時間短縮剤
JP7473283B1 (ja) 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
JP2011069718A (ja) 血液凝固反応における血液凝固時間延長剤
CN115219486B (zh) 一种肝素和低分子肝素抗Xa活性的检测试剂盒及其非疾病诊断检测方法
JP2013145238A (ja) 血液凝固反応における血液凝固時間延長剤
JP2014197019A (ja) 血液凝固反応における血液凝固時間延長剤
Halperin et al. Evaluation of the bicarbonate buffer system
CN116026761A (zh) 抗Xa发色底物法测定试剂盒
JP3181750B2 (ja) イオンセンサ用寿命判定液及び寿命判定方法
WO1997019350A1 (en) A method and a rinse solution used in operating instruments analyzing protein-containing biological liquids

Legal Events

Date Code Title Description
AS Assignment

Owner name: SEKISUI MEDICAL CO., LTD., JAPAN

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:MORIKAWA, CHIZURU;NAKAMURA, REMI;YAMAMOTO, MITSUAKI;SIGNING DATES FROM 20110804 TO 20110817;REEL/FRAME:027282/0722

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION