WO2012043577A1 - Reversing agent for anticoagulants - Google Patents

Reversing agent for anticoagulants Download PDF

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WO2012043577A1
WO2012043577A1 PCT/JP2011/072106 JP2011072106W WO2012043577A1 WO 2012043577 A1 WO2012043577 A1 WO 2012043577A1 JP 2011072106 W JP2011072106 W JP 2011072106W WO 2012043577 A1 WO2012043577 A1 WO 2012043577A1
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blood coagulation
carbonyl
compound
coagulation factor
complex
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French (fr)
Japanese (ja)
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義行 森島
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第一三共株式会社
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    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • A61K31/444—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring heteroatom, e.g. amrinone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A61K38/482—Serine endopeptidases (3.4.21)
    • A61K38/4846—Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/02—Antidotes
    • 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
    • A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents

Definitions

  • the present invention relates to a reverse method using a reverse agent and / or reverse agent that reverses the anticoagulant action of an activated blood coagulation factor X inhibitor.
  • ⁇ Heparins, warfarins, etc. which are anticoagulants that suppress blood coagulation, are known as therapeutic agents for thrombosis and embolism.
  • an activated blood coagulation factor X inhibitor that directly acts on and inhibits activated blood coagulation factor X (hereinafter sometimes abbreviated as FXa) [ Hereinafter, it may be abbreviated as FXa inhibitor.
  • FXa inhibitor activated blood coagulation factor X inhibitor
  • FXa inhibitor activated blood coagulation factor X inhibitor that directly acts on and inhibits activated blood coagulation factor X
  • neutralizing agents or reverse agents for anticoagulant action of anticoagulants for example, protamine for heparins (protamine sulfate), vitamin K1 for warfarin and the like are known (for example, see Non-patent Documents 1 and 2).
  • protamine for heparins protamine sulfate
  • vitamin K1 for warfarin for example, see Non-patent Documents 1 and 2.
  • a non-specific hemostatic agent fresh frozen plasma (FFP) is known, but there is an optimal neutralizing agent or neutralizing method that can be applied depending on the individual anticoagulant. It is also known to be different.
  • FFP fresh frozen plasma
  • the neutralizing agent of the action varies depending on the action mechanism of the anticoagulant, the provision of an optimal neutralizing agent or neutralizing method for each drug having an anticoagulant action is in the treatment of thrombosis / embolism. It is important as a means of treatment in case of drug overdose and serious bleeding.
  • pharmacologically acceptable Salts or their hydrates have a strong direct inhibitory effect on FXa and prevent thrombosis / embolism, particularly deep vein thrombosis (DVT), venous thromboembolism (Venous) Prevention of thrombosis and embolism based on Thromboembolis (VTE) and non-valvular atrial fibrillation (NVAF) Therapeutic agent, as stroke prevention agents in patients with atrial fibrillation, usefulness in clinical trials it has been demonstrated (see, for
  • low-molecular-weight FXa inhibitors are useful for other anticoagulant thrombosis / embolism such as deep vein thrombosis, venous thrombosis / embolism (VTE), and non-valvular atrial fibrillation (NVAF).
  • VTE deep vein thrombosis
  • NVAF non-valvular atrial fibrillation
  • Thrombosis / embolism based, stroke prevention in patients with atrial fibrillation, cerebral infarction, cerebral embolism, myocardial infarction, angina pectoris, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS), Buerger's disease, generalized intravascular Expected to have therapeutic effects on coagulation syndrome, thrombus formation after artificial valve / joint replacement, thrombus formation and re-occlusion after revascularization, multiple organ failure (MODS), thrombus formation during extracorporeal circulation, or blood coagulation during blood collection (For example, refer nonpatent literature 6).
  • Compound (1), a pharmacologically acceptable salt, or a hydrate thereof has a low risk of bleeding at a dose for preventing or treating a disease in clinical trials.
  • a bleeding accident occurs due to, for example, excessive administration of the compound (1), a pharmacologically acceptable salt, or a hydrate thereof, a reverse agent and / or reverse that reverses the anticoagulant action of the drug.
  • the method is not known.
  • the subject of this invention is the reverse agent and / or reverse which reverses the prolonged blood coagulation time at the time of the overdose of compound (1), a pharmacologically acceptable salt, or those hydrates, or at the time of serious bleeding It is to provide a way to do.
  • the present inventors have studied various drugs capable of neutralizing the anticoagulant action of the compound (1), pharmacologically acceptable salts, or hydrates thereof, and prothrombin in an in vitro test using human plasma.
  • Dry human blood coagulation factor IX complex in which blood coagulation time prolonged by compound (1) is used as a hemophilia treatment agent or a blood coagulation factor IX deficiency treatment agent using time (PT) as an index [PPSB®-HT], dried human blood coagulation factor antibody bypassing active complex [Feiba®], and recombinant active factor VII (rFVIIa) are found to be shortened in a concentration-dependent manner
  • PT blood coagulation factor IX deficiency treatment agent
  • PT blood coagulation factor antibody bypassing active complex
  • rFVIIa recombinant active factor VII
  • the present invention provides the following (1) to (4).
  • (1a) a neutralizing agent for the coagulation action of an oral anticoagulant, A neutralizing agent which is one or more selected from a dried human blood coagulation factor antibody bypass activity complex, a dried human blood coagulation factor IX complex, and a recombinant recombinant factor VII.
  • FXa inhibitor is represented by the following formula (1)
  • N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4 which is an anticoagulant, for example, an FXa inhibitor -[(Dimethylamino) carbonyl] -2- ⁇ [(5-methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino ⁇ cyclohexyl) ethanediamide
  • an anticoagulant for example, an FXa inhibitor -[(Dimethylamino) carbonyl] -2- ⁇ [(5-methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino ⁇ cyclohexyl) ethanediamide
  • a reverse agent that reverses the anticoagulant effect of a drug when a pharmacologically acceptable salt or hydrate thereof increases the risk of bleeding due to misadministration or overdose
  • FIG. 4 is a graph showing the influence on prothrombin time (PT) by each of the dried human blood coagulation factor IX complex [PPSB (registered trademark) -HT] and compound (1) alone and in combination, and the results of Example 2
  • FIG. 5 is a graph showing the effect on prothrombin time (PT) by each of recombinant recombinant active factor VII (rFVIIa) and compound (1) alone and in combination, and the results of Example 3.
  • oral anticoagulant means a direct-acting low-molecular oral FXa inhibitor.
  • compound (1) a compound represented by the above formula (1) [hereinafter sometimes abbreviated as compound (1)] is preferable, and compound (1) is It may be a free form (free base) or a hydrate thereof, or may be a pharmacologically acceptable salt or a hydrate of a salt.
  • the pharmacologically acceptable salt of the compound represented by the formula (1) includes hydrochloride, sulfate, hydrobromide, hydroiodide, phosphate, nitrate, benzoate, methanesulfonic acid. Salt, 2-hydroxyethanesulfonate, p-toluenesulfonate, acetate, propanoate, oxalate, malonate, succinate, glutarate, adipate, tartrate, maleate , Fumarate, malate, mandelate and the like.
  • the salt of the compound represented by the formula (1) is preferably hydrochloride or p-toluenesulfonate; p-Toluenesulfonate is particularly preferred.
  • N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2- ⁇ [(5-Methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino ⁇ cyclohexyl) ethanediamide
  • the free base (free form) of “FXa inhibitor” means “acid” of the above-mentioned salt (acid addition salt) and / or acid addition salt forming a hydrate with “FXa inhibitor”. And the compound excluding “water” in the hydrate.
  • the free base (free form) of the compound (1a) is the following formula (1)
  • the dose of the compound (1) represented by the formula (1) is the dose when converted into the compound (1) which is a free base (free form).
  • the FXa inhibitor N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2- ⁇ [(5-methyl- 4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino ⁇ cyclohexyl) ethanediamide / p-toluenesulfonate / monohydrate (compound 1a) Is preferably 5 mg to 90 mg as the free base (free form) [the above compound (1)]; 15 mg to 60 mg is more preferable.
  • the FXa inhibitor N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2- ⁇ [(5-Methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino ⁇ cyclohexyl) ethanediamide, a salt thereof or a hydrate thereof Is administered to a patient in a dosage form such as a tablet together with a pharmacologically acceptable additive.
  • the FXa inhibitor N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2 contained in the administration formulation - ⁇ [(5-Methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino ⁇ cyclohexyl) ethanediamide free base (free form)
  • the content of (1)] is preferably 15 mg, 30 mg, 45 mg and 60 mg; particularly preferably 15 mg, 30 mg and 60 mg.
  • neutralizing agent or “reverse agent” in the present invention means an agent that neutralizes or reverses the anticoagulant action of the anticoagulant. Neutralizing or reversing the action may be understood as detoxifying side effects which are generally undesirable for the human body due to the action of drugs.
  • Anticoagulant “neutralizing agents” or “reverse agents” include dried human blood coagulation factor antibody bypass activity complex [Feiba®], dried human blood coagulation factor IX complex [PPSB®].
  • a dry human blood coagulation factor antibody bypassing active complex [Feiba®]
  • a dry human blood coagulation factor IX complex [PPSB®] — HT]
  • recombinant recombinant factor VII rFVIIa
  • the reverse action of the anticoagulant action in this specification was evaluated using prothrombin time (PT) and activated partial thromboplastin time (APTT) as indices.
  • the clotting time was measured using an automatic blood clotting time measuring machine (Amelung KC-10A micro (Trinity Biotech Plc.)).
  • thrombosis / embolism examples include the following “cerebral infarction, cerebral embolism, myocardial infarction, angina, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS), Buerger's disease, deep vein thrombosis, Generalized intravascular coagulation syndrome, thrombus formation after prosthetic valve / joint replacement, thrombus formation and reocclusion after revascularization, multiple organ failure (MODS), thrombus formation during extracorporeal circulation or blood coagulation during blood collection, venous thrombus -One or more diseases selected from the group consisting of "embolism (VTE) and non-valvular atrial fibrillation (NVAF)” or the like, or thrombosis / embolism based on those diseases; It is not limited to.
  • VTE embolism
  • NVAF non-valvular atrial fibrillation
  • Thromboembolism includes the following cerebral infarction, cerebral embolism, myocardial infarction, angina pectoris, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS), Buerger's disease, deep vein thrombosis, generalized intravascular coagulation syndrome Thrombus formation after prosthetic valve / joint replacement, Thrombus formation and reocclusion after revascularization, Multiple organ failure (MODS), Thrombus formation during extracorporeal circulation or Blood coagulation during blood collection, Venous thrombosis / embolism (VTE), And thrombosis / embolism based on non-valvular atrial fibrillation (NVAF); Preferred is thrombosis / embolism based on venous thrombosis / embolism (VTE) and non-valvular atrial fibrillation (NVAF).
  • VTE venous thrombosis / embolism
  • NVAF non-valvular atrial fibrill
  • formulation means an oral dosage form, together with an active ingredient and a pharmacologically acceptable additive, for example, in a dosage form such as a tablet, granule, powder, capsule or syrup. Be administered.
  • pharmacologically acceptable additive include an excipient, a disintegrant, a binder, a fluidizing agent, a lubricant, a coloring agent, and a brightening agent.
  • Feiba registered trademark
  • distilled water On the day of the experiment, Feiba (registered trademark) was dissolved in distilled water to prepare a 50 U / mL solution. This 50 U / mL solution was diluted with physiological saline to prepare solutions with concentrations of 15, 5, and 1.5 U / mL, and stored in ice for use in the test.
  • Recombinant active factor VII (rFVIIa) solution Recombinant active factor VII (rFVIIa) (NovoSeven (registered trademark); Lot No. X1002) was purchased from Novo Nordisk Pharma Co., Ltd. did.
  • rFVIIa was dissolved in distilled water to prepare a 0.6 mg / mL solution.
  • This 0.6 mg / mL solution was diluted with physiological saline to prepare solutions having a concentration of 10,000, 3000, and 1000 ng / mL, and stored in ice for use in the test.
  • Example 1 Human (Pooled) plasma was thawed in a 37 ° C. water bath and stored in ice for use in experiments. 45 ⁇ L of the solution of compound (1) (16.7 and 33.3 ⁇ g / mL) was added to 4455 ⁇ L of plasma. For control, 45 ⁇ L of 5% DMSO-saline solution was added to plasma.
  • PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.).
  • HemosIL PT-fibrinogen HS PLUS (Instrumentation Laboratory, Lot No. N0298458) was added to start coagulation, and the coagulation time was measured.
  • Human (Pooled) plasma was thawed in a 37 ° C. water bath and stored in ice for use in experiments.
  • 45 ⁇ L of the solution of compound (1) (16.7 and 33.3 ⁇ g / mL) was added to 4455 ⁇ L of plasma.
  • 45 ⁇ L of 5% DMSO-saline solution was added to plasma.
  • PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.) As follows.
  • Example 1 the PT of the control group as a control was 18.4 ⁇ 0.2 seconds (mean value ⁇ standard error, the same applies hereinafter).
  • Addition of 150 or 300 ng / mL concentration of compound (1) significantly extended PT to 31.5 ⁇ 0.2 seconds or 43.7 ⁇ 0.3 seconds, respectively.
  • Single administration (0.15, 0.5, and 1.5 U / mL) of dry human blood coagulation factor antibody bypass activity complex [Feiba®] shortened PT in a concentration-dependent manner.
  • Feiba® significantly shortened the PT prolongation caused by the addition of the compound (1) in a concentration-dependent manner.
  • the PT prolongation caused by the addition of 150 and 300 ng / mL of compound (1) is 19.5 ⁇ 0.1 seconds and The time was shortened to 25.0 ⁇ 0.1 seconds.
  • Example 2 The results of Example 2 are summarized in FIG.
  • the PT of the control group as a control was 17.8 ⁇ 0.0 seconds.
  • the addition of 150 or 300 ng / mL of compound (1) significantly extended PT to 30.7 ⁇ 0.2 seconds or 42.5 ⁇ 0.4 seconds, respectively.
  • [PPSB®-HT] (0.15, 0.5, and 1.5 U / mL) significantly shortened the PT prolongation due to the addition of Compound (1) in a concentration-dependent manner.
  • the PT extension with the addition of 150 and 300 ng / mL of compound (1) is 23.7 ⁇ 0.1 seconds and The time was shortened to 31.6 ⁇ 0.4 seconds.
  • Example 3 The results of Example 3 are summarized in FIG.
  • the PT of the control group as a control was 17.9 ⁇ 0.2 seconds.
  • the addition of 150 or 300 ng / mL of compound (1) significantly extended PT to 30.7 ⁇ 0.2 seconds or 43.1 ⁇ 0.3 seconds, respectively.
  • PT administration was significantly and concentration-dependently shortened by the single administration of recombinant recombinant factor VII (rFVIIa) (100, 300, and 1000 ng / mL).
  • rFVIIa recombinant recombinant factor VII
  • Administration of rFVIIa 100, 300, and 1000 ng / mL significantly shortened the PT prolongation caused by compound (1) in a concentration-dependent manner.
  • the PT prolongation by the addition of 150 and 300 ng / mL of compound (1) is 17.3 ⁇ 0.2 seconds and 24.2 ⁇ 0.3 seconds, respectively.
  • the plasma concentration of the blood coagulation factor VIII preparation at the clinical dose is estimated to be about 0.44 U / mL [package insert of Coordinate (R) FS]. In this study, the concentration was 0.001 to 6.7 U / mL Was used.
  • Compound (1) blood coagulation factor IX preparation [Chris machine (registered trademark) -M] is added to human plasma, and activated partial thromboplastin time (APTT) is measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, (Thrinity Biotech Plc.).
  • the plasma concentration is estimated to be about 0.47 U / mL when Chris Machine (registered trademark) -M is administered at 50 U / kg (near clinical dose) [Chris Machine (registered trademark) -M package insert]. In this case, a concentration of 0.001 to 6.7 U / mL was used.
  • Rats were anesthetized with thiopentasodium (100 mg / kg, ip) 15 minutes after oral administration, and then fresh rat plasma (30 mL / kg / 2h: equivalent to 3 times the clinical dose) 30 minutes after oral administration was continuously administered into the jugular vein.
  • physiological saline (30 mL / kg / 2h) was continuously administered into the jugular vein.
  • Citrated blood was collected from the jugular vein 2 hours after the start of continuous intravenous infusion of fresh plasma. Plasma was separated from the collected blood, and PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.).
  • Compound (1) extended PT by 2.14 times.
  • Fresh plasma (FF) reduced PT prolongation by compound (1) from 2.14 times to 1.66 times at a dose 3 times higher than the clinical dose.
  • Comparative Example 4 Rats fasted overnight were orally administered compound (1) (10 mg / kg) suspended in 0.5% methylcellulose, and vitamin K2 (0.1 and 1 mg / kg) or distilled water was orally administered 30 minutes later. did.
  • Vitamin K2 0.1 and 1 mg / kg or distilled water was orally administered 30 minutes later. did.
  • One hour and 45 minutes after vitamin K2 administration the rats were anesthetized with thiopentasodium (100 mg / kg, ip), and citrated blood was collected from the inferior vena cava 15 minutes later. Plasma was separated from the collected blood, and PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.).
  • Compound (1) prolonged PT by 1.34 times 2.5 hours after oral administration. Vitamin K2 did not affect PT prolonged by compound (1).
  • Rats fasted overnight were anesthetized with thiopental sodium (0.1 g / kg, ip). From 15 minutes after anesthesia to the end of the experiment, 5% glucose solution or compound (1) was continuously administered from the jugular vein (1 mg / kg / h), and the bleeding time was measured 2 hours after the start of administration. Saline solution or tranexamic acid was injected over 2 minutes from the femoral vein 8 minutes before the bleeding time measurement. The bleeding time was measured by the plantar template method or the tail template method.
  • the rat was placed in a supine position, and a 5 mm long incision was made with a cutter knife in the sole of the right hind limb.
  • the blood was adsorbed on the filter paper every 30 seconds, and the time until blood did not bleed on the filter paper was measured up to 30 minutes. If the hemostasis was not stopped within 30 minutes, the bleeding time was 30 minutes.
  • the rat was placed in a supine position, and a 1 mm deep incision was made with a razor into the arterial portion 4 cm from the tip of the tail.
  • the blood was adsorbed on the filter paper every 15 seconds, and the time until blood did not bleed on the filter paper was measured up to 30 minutes.
  • plantar bleeding time was significantly prolonged by administration of compound (1).
  • administration of 30, 100 and 300 mg / kg tranexamic acid showed no inhibitory effect.
  • PT was extended 1.36 times, and the same PT extension was observed in the 30 mg / kg tranexamic acid combination group.
  • administration of compound (1) significantly prolonged rat tail bleeding time (the rate of extension was 1.77 times).
  • administration of 30 and 100 mg / kg tranexamic acid showed no inhibitory effect.
  • the compound (1) prolongs PT by 1.33 times, and the PT in the tranexamic acid combination group similarly prolongs PT, and no inhibitory effect on PT prolongation by administration of compound (1) was observed.
  • tranexamic acid showed a reverse action of compound (1) at doses of 30, 100, and 300 mg / kg, with respect to prolonged bleeding time by compound (1) using rat plantar and tail bleeding time models. There wasn't.
  • the present invention relates to a reverse agent that reverses the anticoagulant action of a drug in the event that a bleeding accident occurs due to, for example, excessive administration of compound (1), a pharmacologically acceptable salt, or a hydrate thereof. Alternatively, it can be used as a reverse method.

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Abstract

The problem addressed by the present invention is the provision of: a reversing agent that reverses the anticoagulant effects of an FXa inhibitor; and/or a reversal method using said reversing agent. [Solution] A reversing agent for an oral anticoagulant, said reversing agent causing clotting. Said clotting agent is one or more of the following: a dried complex that has the effect of circumventing antibodies for a human blood coagulation factor; a dried complex of human blood coagulation factor IX; and recombinant activated factor VII.

Description

抗凝固作用のリバース剤Anticoagulant reverse agent
　本発明は、活性化血液凝固第Ｘ因子阻害薬の抗凝固作用をリバースする、リバース剤及び／又はリバース剤を用いたリバース方法に関する。 The present invention relates to a reverse method using a reverse agent and / or reverse agent that reverses the anticoagulant action of an activated blood coagulation factor X inhibitor.
　血栓・塞栓症の治療薬として血液の凝固を抑える抗凝固薬であるヘパリン類、ワルファリン等が知られている。また、新たな作用メカニズムの抗凝固薬として、活性化された血液凝固第Ｘ因子［以下、ＦＸａと略する場合もある］に直接作用して阻害する、活性化血液凝固第Ｘ因子阻害薬［以下、ＦＸａ阻害薬と略する場合もある］等も現在開発されている。これらの抗凝固薬は、その作用メカニズムである血液凝固阻害作用に基づき、副作用として出血のリスクを持つことが知られている。したがって、抗凝固薬の過剰投与時、又は重大な出血時等、投与薬物の血液凝固阻害作用をリバース（中和）し、延長された凝固時間を正常値に戻す薬剤、又は方法が必要である。 <Heparins, warfarins, etc., which are anticoagulants that suppress blood coagulation, are known as therapeutic agents for thrombosis and embolism. In addition, as an anticoagulant with a new mechanism of action, an activated blood coagulation factor X inhibitor that directly acts on and inhibits activated blood coagulation factor X (hereinafter sometimes abbreviated as FXa) [ Hereinafter, it may be abbreviated as FXa inhibitor. These anticoagulants are known to have a risk of bleeding as a side effect based on the blood coagulation inhibitory action which is the action mechanism. Therefore, there is a need for a drug or method that reverses (neutralizes) the blood coagulation inhibitory action of the administered drug and returns the prolonged clotting time to a normal value, such as when an anticoagulant is overdosed or severe bleeding. .
　抗凝固薬の抗凝固作用の中和剤又はリバース剤として、例えば、ヘパリン類に対するプロタミン（硫酸プロタミン）、また、ワルファリンに対するビタミンＫ１等が知られている（例えば、非特許文献１～２参照）。また、非特異的な止血剤としては新鮮凍結血漿（ＦＦＰ：ｆｒｅｓｈ　ｆｒｏｚｅｎ　ｐｌａｓｍａ）等が知られているが、個々の抗凝固薬によって、適用可能で、かつ最適な中和剤又は中和方法が異なることも知られている。このように、抗凝固薬の作用メカニズムによって作用の中和剤が異なることから、個々の抗凝固作用を持つ薬物の最適な中和剤又は中和方法の提供は、血栓・塞栓症の治療において、薬剤の過剰投与や重大な出血の際の治療手段として重要である。 As neutralizing agents or reverse agents for anticoagulant action of anticoagulants, for example, protamine for heparins (protamine sulfate), vitamin K1 for warfarin and the like are known (for example, see Non-patent Documents 1 and 2). . In addition, as a non-specific hemostatic agent, fresh frozen plasma (FFP) is known, but there is an optimal neutralizing agent or neutralizing method that can be applied depending on the individual anticoagulant. It is also known to be different. Thus, since the neutralizing agent of the action varies depending on the action mechanism of the anticoagulant, the provision of an optimal neutralizing agent or neutralizing method for each drug having an anticoagulant action is in the treatment of thrombosis / embolism. It is important as a means of treatment in case of drug overdose and serious bleeding.
　下記式（１） The following formula (1)
Figure JPOXMLDOC01-appb-C000003
Figure JPOXMLDOC01-appb-C000003
で表される、Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド［以下、化合物（１）と略する場合もある）、薬理上許容される塩、又はそれらの水和物は、強力なＦＸａの直接阻害作用を有し、血栓・塞栓症の予防薬、特に深部静脈血栓症（Ｄｅｅｐ　ｖｅｉｎ　ｔｈｒｏｍｂｏｓｉｓ：ＤＶＴ）、静脈血栓・塞栓症（Ｖｅｎｏｕｓ　Ｔｈｒｏｍｂｏｅｍｂｏｌｉｓｍ：ＶＴＥ）及び非弁膜性心房細動（Ｎｏｎ－Ｖａｌｖｕｌａｒ　Ａｔｒｉａｌ　Ｆｉｂｒｉｌｌａｔｉｏｎ：ＮＶＡＦ）に基づく血栓・塞栓症の予防的治療薬、心房細動患者における脳卒中予防薬として、臨床試験での有用性が証明されてきている（例えば、特許文献１～２及び非特許文献３～５参照）。さらに、低分子ＦＸａ阻害薬は、上記以外の抗凝固薬血栓・塞栓症、例えば、深部静脈血栓症、静脈血栓・塞栓症（ＶＴＥ）、及び非弁膜性心房細動（ＮＶＡＦ）等の疾患に基づく血栓・塞栓症、心房細動患者における脳卒中予防の他、脳梗塞、脳塞栓、心筋梗塞、狭心症、肺梗塞、肺塞栓、急性冠動脈症候群（ＡＣＳ）、バージャー病、汎発性血管内凝固症候群、人工弁／関節置換後の血栓形成、血行再建後の血栓形成及び再閉塞、多臓器不全（ＭＯＤＳ）、体外循環時の血栓形成又は採血時の血液凝固等への治療効果が期待されることが報告されている（例えば、非特許文献６参照）。また、化合物（１）、薬理上許容される塩、又はそれらの水和物は、臨床試験において、疾患の予防・治療投与用量における出血リスクが少ないことも報告されている。しかし、化合物（１）、薬理上許容される塩、又はそれらの水和物が、例えば過剰投与等によって万が一に出血事故が発生した場合、薬物の抗凝固作用をリバースするリバース剤及び／又はリバース方法は知られていない。 N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl- 4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide [hereinafter sometimes abbreviated as compound (1)), pharmacologically acceptable Salts or their hydrates have a strong direct inhibitory effect on FXa and prevent thrombosis / embolism, particularly deep vein thrombosis (DVT), venous thromboembolism (Venous) Prevention of thrombosis and embolism based on Thromboembolis (VTE) and non-valvular atrial fibrillation (NVAF) Therapeutic agent, as stroke prevention agents in patients with atrial fibrillation, usefulness in clinical trials it has been demonstrated (see, for example, to 5 Patent Documents 1-2 and Non-Patent Document 3). Furthermore, low-molecular-weight FXa inhibitors are useful for other anticoagulant thrombosis / embolism such as deep vein thrombosis, venous thrombosis / embolism (VTE), and non-valvular atrial fibrillation (NVAF). Thrombosis / embolism based, stroke prevention in patients with atrial fibrillation, cerebral infarction, cerebral embolism, myocardial infarction, angina pectoris, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS), Buerger's disease, generalized intravascular Expected to have therapeutic effects on coagulation syndrome, thrombus formation after artificial valve / joint replacement, thrombus formation and re-occlusion after revascularization, multiple organ failure (MODS), thrombus formation during extracorporeal circulation, or blood coagulation during blood collection (For example, refer nonpatent literature 6). In addition, it has been reported that Compound (1), a pharmacologically acceptable salt, or a hydrate thereof has a low risk of bleeding at a dose for preventing or treating a disease in clinical trials. However, if a bleeding accident occurs due to, for example, excessive administration of the compound (1), a pharmacologically acceptable salt, or a hydrate thereof, a reverse agent and / or reverse that reverses the anticoagulant action of the drug. The method is not known.
国際公開第２００３／０００６８０号パンフレットInternational Publication No. 2003/000680 Pamphlet 国際公開第２００４／０５８７１５号パンフレットInternational Publication No. 2004/058715 Pamphlet
　本発明の課題は、化合物（１）、薬理上許容される塩、又はそれらの水和物の過剰投与時又は重大な出血時において、延長された血液凝固時間をリバースするリバース剤及び／又はリバースする方法を提供することにある。 The subject of this invention is the reverse agent and / or reverse which reverses the prolonged blood coagulation time at the time of the overdose of compound (1), a pharmacologically acceptable salt, or those hydrates, or at the time of serious bleeding It is to provide a way to do.
　本発明者らは、化合物（１）、薬理上許容される塩、又はそれらの水和物の抗凝固作用を中和可能な薬剤を種々検討し、ヒト血漿を用いたｉｎ　ｖｉｔｒｏ試験において、プロトロンビン時間（ＰＴ）等を指標とし、化合物（１）によって延長された血液凝固時間を、血友病治療剤又は血液凝固第ＩＸ因子欠乏症治療剤として用いられている乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］、乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］、及び遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）が濃度依存的に短縮することを見出して本発明を完成した。 The present inventors have studied various drugs capable of neutralizing the anticoagulant action of the compound (1), pharmacologically acceptable salts, or hydrates thereof, and prothrombin in an in vitro test using human plasma. Dry human blood coagulation factor IX complex in which blood coagulation time prolonged by compound (1) is used as a hemophilia treatment agent or a blood coagulation factor IX deficiency treatment agent using time (PT) as an index [PPSB®-HT], dried human blood coagulation factor antibody bypassing active complex [Feiba®], and recombinant active factor VII (rFVIIa) are found to be shortened in a concentration-dependent manner The present invention has been completed.
　すなわち、本発明は下記の（１）～（４）を提供するものである。
（１ａ）：経口抗凝固薬の凝固作用の中和剤であって、
乾燥人血液凝固因子抗体迂回活性複合体、乾燥人血液凝固第ＩＸ因子複合体、及び、遺伝子組換え活性型第ＶＩＩ因子から選ばれる１以上である中和剤。
（１ｂ）：ＦＸａ阻害薬の凝固作用の中和剤であって、
乾燥人血液凝固因子抗体迂回活性複合体、乾燥人血液凝固第ＩＸ因子複合体、及び、遺伝子組換え活性型第ＶＩＩ因子から選ばれる１以上である中和剤。
（１）：ＦＸａ阻害薬の凝固作用のリバース剤であって、
乾燥人血液凝固因子抗体迂回活性複合体、乾燥人血液凝固第ＩＸ因子複合体、及び、遺伝子組換え活性型第ＶＩＩ因子から選ばれる１以上であるリバース剤。
（２ａ）：ＦＸａ阻害薬が、下記式（１） That is, the present invention provides the following (1) to (4).
(1a): a neutralizing agent for the coagulation action of an oral anticoagulant,
A neutralizing agent which is one or more selected from a dried human blood coagulation factor antibody bypass activity complex, a dried human blood coagulation factor IX complex, and a recombinant recombinant factor VII.
(1b): a neutralizing agent for coagulation of an FXa inhibitor,
A neutralizing agent which is one or more selected from a dried human blood coagulation factor antibody bypass activity complex, a dried human blood coagulation factor IX complex, and a recombinant recombinant factor VII.
(1): a reverse agent of the coagulation action of an FXa inhibitor,
A reverse agent that is one or more selected from a dried human blood coagulation factor antibody bypass activity complex, a dried human blood coagulation factor IX complex, and a recombinant recombinant factor VII.
(2a): FXa inhibitor is represented by the following formula (1)
Figure JPOXMLDOC01-appb-C000004
Figure JPOXMLDOC01-appb-C000004
で表されるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド、薬理上許容される塩、又はそれらの水和物である、（１ｂ）に記載の中和剤。
（２）：ＦＸａ阻害薬が、下記式（１） N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4 , 5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, a pharmacologically acceptable salt, or a hydrate thereof, (1b) The neutralizing agent described in 1.
(2): FXa inhibitor is represented by the following formula (1)
Figure JPOXMLDOC01-appb-C000005
Figure JPOXMLDOC01-appb-C000005
で表されるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド、薬理上許容される塩、又はそれらの水和物である、（１）に記載のリバース剤。
（３ａ）：ＦＸａ阻害薬が、下記式（１ａ） N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4 , 5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, a pharmacologically acceptable salt, or a hydrate thereof, (1) The reverse agent described in 1.
(3a): FXa inhibitor is represented by the following formula (1a)
Figure JPOXMLDOC01-appb-C000006
Figure JPOXMLDOC01-appb-C000006
で表されるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド・ｐ－トルエンスルホン酸塩・１水和物である、（１ｂ）に記載の中和剤。
（３）：ＦＸａ阻害薬が、下記式（１ａ） N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4 , 5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, p-toluenesulfonate, monohydrate, described in (1b) Neutralizer.
(3): FXa inhibitor is represented by the following formula (1a)
Figure JPOXMLDOC01-appb-C000007
Figure JPOXMLDOC01-appb-C000007
で表されるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド・ｐ－トルエンスルホン酸塩・１水和物である、（１）に記載のリバース剤。
（４ａ）：乾燥人血液凝固因子抗体迂回活性複合体、乾燥人血液凝固第ＩＸ因子複合体、及び、遺伝子組換え活性型第ＶＩＩ因子から選ばれる１以上を用いることを特徴とする、Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド、薬理上許容される塩、又はそれらの水和物のＦＸａ阻害作用を中和する方法。
（４）：乾燥人血液凝固因子抗体迂回活性複合体、乾燥人血液凝固第ＩＸ因子複合体、及び、遺伝子組換え活性型第ＶＩＩ因子から選ばれる１以上を用いることを特徴とする、Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド、薬理上許容される塩、又はそれらの水和物のＦＸａ阻害作用をリバースする方法。 N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4 , 5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, p-toluenesulfonate, monohydrate, described in (1) Reverse agent.
(4a): One or more selected from dry human blood coagulation factor antibody bypass activity complex, dry human blood coagulation factor IX complex, and recombinant recombinant factor VII, N 1 -(5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4,5,6,7 A method for neutralizing the FXa inhibitory action of tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, pharmacologically acceptable salts, or hydrates thereof.
(4): One or more selected from dry human blood coagulation factor antibody bypass active complex, dry human blood coagulation factor IX complex, and recombinant recombinant factor VII, N 1 -(5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4,5,6,7 A method for reversing the FXa inhibitory action of tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, pharmacologically acceptable salts, or hydrates thereof.
　本発明によれば、血栓・塞栓治療において、抗凝固薬、例えばＦＸａ阻害薬である、Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド、薬理上許容される塩、又はそれらの水和物が、誤投与や、過剰投与等によって出血リスクが高まったとき、又は重大な出血事故が発生した時に、薬物の抗凝固作用をリバースするリバース剤及び／又はリバースする方法が提供される。 According to the present invention, in the treatment of thrombosis / embolism, N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4, which is an anticoagulant, for example, an FXa inhibitor -[(Dimethylamino) carbonyl] -2-{[(5-methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, A reverse agent that reverses the anticoagulant effect of a drug when a pharmacologically acceptable salt or hydrate thereof increases the risk of bleeding due to misadministration or overdose, or when a serious bleeding accident occurs And / or a reverse method is provided.
：乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］及び化合物（１）のそれぞれ単独及び併用によるプロトロンビン時間（ＰＴ）への影響を示した図であり、実施例１の結果を示す図である。: It is the figure which showed the influence on prothrombin time (PT) by the dry human blood coagulation factor antibody detour active complex [Feiba (registered trademark)] and the compound (1) each alone and in combination. FIG. ：乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］及び化合物（１）のそれぞれ単独及び併用によるプロトロンビン時間（ＰＴ）への影響を示した図であり、実施例２の結果を示す図である。FIG. 4 is a graph showing the influence on prothrombin time (PT) by each of the dried human blood coagulation factor IX complex [PPSB (registered trademark) -HT] and compound (1) alone and in combination, and the results of Example 2 FIG. ：遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）及び化合物（１）のそれぞれ単独及び併用によるプロトロンビン時間（ＰＴ）への影響を示した図であり、実施例３の結果を示す図である。FIG. 5 is a graph showing the effect on prothrombin time (PT) by each of recombinant recombinant active factor VII (rFVIIa) and compound (1) alone and in combination, and the results of Example 3.
　以下に本発明を詳細に説明する。 The present invention will be described in detail below.
　本明細書における「経口抗凝固薬」とは、直接作用型の低分子経口ＦＸａ阻害薬を意味する。 In the present specification, “oral anticoagulant” means a direct-acting low-molecular oral FXa inhibitor.
　本明細書における「ＦＸａ阻害薬」の具体的な例としては、上記の式（１）で表される化合物［以下、化合物（１）と略する場合がある］が好ましく、化合物（１）はフリー体（遊離塩基）、その水和物でもよく、また薬理学上許容される塩、塩の水和物であってもよい。 As a specific example of “FXa inhibitor” in the present specification, a compound represented by the above formula (1) [hereinafter sometimes abbreviated as compound (1)] is preferable, and compound (1) is It may be a free form (free base) or a hydrate thereof, or may be a pharmacologically acceptable salt or a hydrate of a salt.
　式（１）で表される化合物の薬理学上許容される塩としては、塩酸塩、硫酸塩、臭化水素酸塩、ヨウ化水素酸塩、燐酸塩、硝酸塩、安息香酸塩、メタンスルホン酸塩、２－ヒドロキシエタンスルホン酸塩、ｐ－トルエンスルホン酸塩、酢酸塩、プロパン酸塩、シュウ酸塩、マロン酸塩、コハク酸塩、グルタル酸塩、アジピン酸塩、酒石酸塩、マレイン酸塩、フマル酸塩、リンゴ酸塩、マンデル酸塩等が挙げられる。 The pharmacologically acceptable salt of the compound represented by the formula (1) includes hydrochloride, sulfate, hydrobromide, hydroiodide, phosphate, nitrate, benzoate, methanesulfonic acid. Salt, 2-hydroxyethanesulfonate, p-toluenesulfonate, acetate, propanoate, oxalate, malonate, succinate, glutarate, adipate, tartrate, maleate , Fumarate, malate, mandelate and the like.
　式（１）で表される化合物の塩としては、塩酸塩、ｐ－トルエンスルホン酸塩が好ましく；
ｐ－トルエンスルホン酸塩が特に好ましい。 The salt of the compound represented by the formula (1) is preferably hydrochloride or p-toluenesulfonate;
p-Toluenesulfonate is particularly preferred.
　下記式（１） The following formula (1)
Figure JPOXMLDOC01-appb-C000008
Figure JPOXMLDOC01-appb-C000008
で表される化合物として好ましいものとして、以下の
Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド；
Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド・塩酸塩；
Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド・ｐ－トルエンスルホン酸塩；及び
Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド・ｐ－トルエンスルホン酸塩・１水和物；
を挙げることができる。 As preferred compounds represented by formula (1), the following N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2- {[(5-Methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide;
N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4,5,6 , 7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide hydrochloride;
N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4,5,6 , 7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide p-toluenesulfonate; and N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridine- 2-yl) carbonyl] amino} cyclohexyl) ethanediamide p-toluenesulfonate monohydrate;
Can be mentioned.
　上記の好ましい化合物の中で、下記式（１ａ） Among the above preferred compounds, the following formula (1a)
Figure JPOXMLDOC01-appb-C000009
Figure JPOXMLDOC01-appb-C000009
で表される、Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド・ｐ－トルエンスルホン酸塩・１水和物［以下、化合物（１ａ）と略する場合がある］が特に好ましい。 N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl- 4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide p-toluenesulfonate monohydrate [hereinafter referred to as compound (1a) Is sometimes abbreviated as].
　「ＦＸａ阻害薬」の遊離塩基（フリー体）とは、「ＦＸａ阻害薬」と、たとえば上記の塩（酸付加塩）、及び／又は水和物を形成している酸付加塩の「酸」及び水和物の「水」を除いた化合物を意味する。例えば、上記化合物（１ａ）の遊離塩基（フリー体）とは、下記式（１） The free base (free form) of “FXa inhibitor” means “acid” of the above-mentioned salt (acid addition salt) and / or acid addition salt forming a hydrate with “FXa inhibitor”. And the compound excluding “water” in the hydrate. For example, the free base (free form) of the compound (1a) is the following formula (1)
Figure JPOXMLDOC01-appb-C000010
Figure JPOXMLDOC01-appb-C000010
で表されるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド（１）を意味する。本明細書中における式（１）で表される化合物（１）の投与量は、すべて遊離塩基（フリー体）である化合物（１）に換算したときの投与量を用いた。 N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4 , 5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide (1). In the present specification, the dose of the compound (1) represented by the formula (1) is the dose when converted into the compound (1) which is a free base (free form).
　ＦＸａ阻害薬であるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド・ｐ－トルエンスルホン酸塩・１水和物（化合物１ａ）の投与量としては、遊離塩基（フリー体）［上記の化合物（１）］として、５ｍｇ～９０ｍｇが好ましく；
１５ｍｇ～６０ｍｇがより好ましい。 The FXa inhibitor N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl- 4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide / p-toluenesulfonate / monohydrate (compound 1a) Is preferably 5 mg to 90 mg as the free base (free form) [the above compound (1)];
15 mg to 60 mg is more preferable.
　経口血液凝固抑制剤として投与する場合、ＦＸａ阻害薬であるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド、その塩又はそれらの水和物を、薬理学的に許容される添加剤とともに、例えば錠剤等の投与剤形で患者に投与される。その投与製剤中に含まれる、ＦＸａ阻害薬であるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミドの遊離塩基（フリー体）［上記の化合物（１）］の含有量としては、１５ｍｇ、３０ｍｇ、４５ｍｇ及び６０ｍｇが好ましく；１５ｍｇ、３０ｍｇ及び６０ｍｇが特に好ましい。 When administered as an oral blood coagulation inhibitor, the FXa inhibitor N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-Methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, a salt thereof or a hydrate thereof Is administered to a patient in a dosage form such as a tablet together with a pharmacologically acceptable additive. The FXa inhibitor N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2 contained in the administration formulation -{[(5-Methyl-4,5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide free base (free form) The content of (1)] is preferably 15 mg, 30 mg, 45 mg and 60 mg; particularly preferably 15 mg, 30 mg and 60 mg.
　本願発明おける「中和剤」又は「リバース剤」としては、抗凝固薬のもつ抗凝固作用を中和又はリバースする薬剤を意味する。作用の中和又はリバースとは、一般的には薬物の作用による、人体にとって好ましくない副作用を解毒すると解されることもある。抗凝固薬の「中和剤」又は「リバース剤」としては、乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］、乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］、遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）、血液凝固第ＶＩＩＩ因子製剤［コージネイト（登録商標）ＦＳ］、血液凝固第ＩＸ因子製剤［クリスマシン（登録商標）－Ｍ］、新鮮凍結血漿（ＦＦＰ：Ｆｒｅｓｈ　Ｆｒｏｚｅｎ　Ｐｌａｓｍａ）、ビタミンＫ（ビタミンＫ１、ビタミンＫ２を包含する）、トラネキサム酸等を挙げることができる。ここで、乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］とは、ＰＰＳＢ（登録商標）－ＨＴ「ニチヤク」の医薬品インタビューファーム［２００９年４月（第３版）］に記載されているように、高力価の血液凝固第ＩＸ因子を含有する製剤であり（２０単位／ｍＬ以上）血液凝固第ＩＸ因子以外に、他のビタミンＫ依存性の血液凝固因子である第ＩＩ、第ＶＩＩ、及び第Ｘ因子を含有する「プロトロンビン複合体濃縮製剤（ＰＣＣ：Ｐｒｏｔｈｒｏｍｂｉｎ　Ｃｏｍｐｌｅｘ　Ｃｏｎｃｅｎｔｒａｔｅ）」を意味する。 The “neutralizing agent” or “reverse agent” in the present invention means an agent that neutralizes or reverses the anticoagulant action of the anticoagulant. Neutralizing or reversing the action may be understood as detoxifying side effects which are generally undesirable for the human body due to the action of drugs. Anticoagulant “neutralizing agents” or “reverse agents” include dried human blood coagulation factor antibody bypass activity complex [Feiba®], dried human blood coagulation factor IX complex [PPSB®]. -HT], recombinant recombinant factor VII (rFVIIa), blood coagulation factor VIII preparation [Cognate (R) FS], blood coagulation factor IX preparation [Chris Machine (R) -M], fresh frozen Examples thereof include plasma (FFP: Fresh Frozen Plasma), vitamin K (including vitamin K1 and vitamin K2), tranexamic acid, and the like. Here, the dried human blood coagulation factor IX complex [PPSB (registered trademark) -HT] refers to PPSB (registered trademark) -HT "Nichiyaku" pharmaceutical interview farm [April 2009 (3rd edition)]. As described, it is a preparation containing high-titer blood coagulation factor IX (20 units / mL or more), in addition to blood coagulation factor IX, other vitamin K-dependent blood coagulation factors It means “Prothrombin Complex Concentrate (PCC)” containing II, Factor VII, and Factor X.
　本願発明における「リバース剤」又は「中和剤」としては、乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］、乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］、及び遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）が好ましく；
乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］及び遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）がより好ましい。 As the “reverse agent” or “neutralizing agent” in the present invention, a dry human blood coagulation factor antibody bypassing active complex [Feiba®], a dry human blood coagulation factor IX complex [PPSB®] — HT], and recombinant recombinant factor VII (rFVIIa) are preferred;
More preferred are dried human blood coagulation factor antibody bypass active complex [Feiba®] and recombinant factor VII (rFVIIa).
　本願明細書における抗凝固作用のリバース作用は、プロトロンビン時間（ＰＴ）及び活性化部分トロンボプラスチン時間（ＡＰＴＴ）を指標として評価した。また、凝固時間は、血液凝固時間自動測定機（Ａｍｅｌｕｎｇ　ＫＣ－１０Ａ　ｍｉｃｒｏ（Ｔｒｉｎｉｔｙ　Ｂｉｏｔｅｃｈ　Ｐｌｃ．）を用いて測定した。 The reverse action of the anticoagulant action in this specification was evaluated using prothrombin time (PT) and activated partial thromboplastin time (APTT) as indices. The clotting time was measured using an automatic blood clotting time measuring machine (Amelung KC-10A micro (Trinity Biotech Plc.)).
　本明細書におけるデータの統計学的有意性の解析は、Ｄｕｎｎｅｔｔ法を用いた。＃＃＃はＰ＜０．００１、＄＄＄はＰ＜０．００１　対　対照群（化合物（１）とリバース剤の非存在）。＊＊＊はＰ＜０．００１　対　化合物（１）群の有意差をそれぞれ示したものである。 The Dunnett method was used for analysis of the statistical significance of the data in this specification. ## is P <0.001, and $$ is P <0.001 vs. control group (compound (1) and absence of reverse agent). *** indicates a significant difference between P <0.001 and the compound (1) group.
　本明細書における、血栓・塞栓症としては、例えば下記の
「脳梗塞、脳塞栓、心筋梗塞、狭心症、肺梗塞、肺塞栓、急性冠動脈症候群（ＡＣＳ）、バージャー病、深部静脈血栓症、汎発性血管内凝固症候群、人工弁／関節置換後の血栓形成、血行再建後の血栓形成及び再閉塞、多臓器不全（ＭＯＤＳ）、体外循環時の血栓形成又は採血時の血液凝固、静脈血栓・塞栓症（ＶＴＥ）及び非弁膜性心房細動（ＮＶＡＦ）」等からなる群より選ばれる１以上の疾患或いはそれら疾患に基づく血栓・塞栓症；を挙げることができるが、なんらこれらの疾患のみに限定されるものではない。 Examples of thrombosis / embolism in the present specification include the following “cerebral infarction, cerebral embolism, myocardial infarction, angina, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS), Buerger's disease, deep vein thrombosis, Generalized intravascular coagulation syndrome, thrombus formation after prosthetic valve / joint replacement, thrombus formation and reocclusion after revascularization, multiple organ failure (MODS), thrombus formation during extracorporeal circulation or blood coagulation during blood collection, venous thrombus -One or more diseases selected from the group consisting of "embolism (VTE) and non-valvular atrial fibrillation (NVAF)" or the like, or thrombosis / embolism based on those diseases; It is not limited to.
　血栓・塞栓症としては、下記の
脳梗塞、脳塞栓、心筋梗塞、狭心症、肺梗塞、肺塞栓、急性冠動脈症候群（ＡＣＳ）、バージャー病、深部静脈血栓症、汎発性血管内凝固症候群、人工弁／関節置換後の血栓形成、血行再建後の血栓形成及び再閉塞、多臓器不全（ＭＯＤＳ）、体外循環時の血栓形成又は採血時の血液凝固、静脈血栓・塞栓症（ＶＴＥ）、及び非弁膜性心房細動（ＮＶＡＦ）に基づく血栓・塞栓症；
が好ましく；静脈血栓・塞栓症（ＶＴＥ）及び非弁膜性心房細動（ＮＶＡＦ）に基づく血栓・塞栓症がより好ましい。 Thromboembolism includes the following cerebral infarction, cerebral embolism, myocardial infarction, angina pectoris, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS), Buerger's disease, deep vein thrombosis, generalized intravascular coagulation syndrome Thrombus formation after prosthetic valve / joint replacement, Thrombus formation and reocclusion after revascularization, Multiple organ failure (MODS), Thrombus formation during extracorporeal circulation or Blood coagulation during blood collection, Venous thrombosis / embolism (VTE), And thrombosis / embolism based on non-valvular atrial fibrillation (NVAF);
Preferred is thrombosis / embolism based on venous thrombosis / embolism (VTE) and non-valvular atrial fibrillation (NVAF).
　本明細書における製剤としては、経口製剤を意味し、有効成分及び薬理学的に許容される添加剤とともに、例えば錠剤、顆粒剤、粉末剤、カプセル剤又はシロップ剤等の投与剤形で患者に投与される。薬理学的に許容される添加剤としては、例えば賦形剤、崩壊剤、結合剤、流動化剤、滑沢剤、着色剤、光沢化剤等を挙げることができる。 As used herein, the term “formulation” means an oral dosage form, together with an active ingredient and a pharmacologically acceptable additive, for example, in a dosage form such as a tablet, granule, powder, capsule or syrup. Be administered. Examples of the pharmacologically acceptable additive include an excipient, a disintegrant, a binder, a fluidizing agent, a lubricant, a coloring agent, and a brightening agent.
　次に実施例を挙げて本発明を詳細に説明するが、本発明は何らこれら実施例に限定されるものではない。 Next, the present invention will be described in detail with reference to examples, but the present invention is not limited to these examples.
　［評価用試験溶液の調製］
　（１）化合物（１）の溶液
試験当日に化合物（１ａ）を秤量し、５％ＤＭＳＯ－生理食塩水溶液で希釈して３３．３μｇ／ｍＬ、及び１６．７μｇ／ｍＬの濃度［遊離塩基である化合物（１）に換算した濃度］の溶液を調製し、遮光条件下、室温で保存して試験に使用した。 [Preparation of test solution for evaluation]
(1) The solution (1a) of compound (1) is weighed and diluted with 5% DMSO-saline solution to give concentrations of 33.3 μg / mL and 16.7 μg / mL [free base. The solution of the concentration converted to the compound (1)] was prepared, stored at room temperature under light-shielding conditions, and used for the test.
　（２）乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］溶液
乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］（ロットＮｏ．ＶＮＦ１Ｈ０４０；有効成分：５００Ｕ）は、バクスター株式会社から購入した。 (2) Dried human blood coagulation factor antibody bypass activity complex [Feiba (registered trademark)] Solution dry human blood coagulation factor antibody bypass activity complex [Feiba (registered trademark)] (Lot No. VNF1H040; active ingredient: 500 U) , Purchased from Baxter Corporation.
　実験の当日にＦｅｉｂａ（登録商標）を蒸留水で溶解し、５０Ｕ／ｍＬ溶液を調製した。この５０Ｕ／ｍＬ溶液を生理食塩水で希釈し、１５、５、及び１．５Ｕ／ｍＬの濃度の溶液を調製し、氷中で保存して試験に用いた。 On the day of the experiment, Feiba (registered trademark) was dissolved in distilled water to prepare a 50 U / mL solution. This 50 U / mL solution was diluted with physiological saline to prepare solutions with concentrations of 15, 5, and 1.5 U / mL, and stored in ice for use in the test.
　（３）乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］溶液
乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］（Ｌｏｔ　Ｎｏ．Ｎ１１７ＤＫ；２００Ｕ）は、日本製薬株式会社から購入した。 (3) Dried human blood coagulation factor IX complex [PPSB (registered trademark) -HT] solution Dried human blood coagulation factor IX complex [PPSB (registered trademark) -HT] (Lot No. N117DK; 200 U) Purchased from Nippon Pharmaceutical Co., Ltd.
　実験の当日に［ＰＰＳＢ（登録商標）－ＨＴ］を蒸留水で溶解し、２０Ｕ／ｍＬ溶液を調製した。この２０Ｕ／ｍＬ溶液を生理食塩水で希釈し、１５、５、及び１．５Ｕ／ｍＬの濃度の溶液を調製し、氷中で保存して試験に用いた。 On the day of the experiment, [PPSB (registered trademark) -HT] was dissolved in distilled water to prepare a 20 U / mL solution. This 20 U / mL solution was diluted with physiological saline to prepare solutions with concentrations of 15, 5, and 1.5 U / mL, and stored in ice for use in the test.
　（４）遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）溶液
遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）（ＮｏｖｏＳｅｖｅｎ（登録商標）；Ｌｏｔ　Ｎｏ．Ｘ１００２）は、ノボ・ノルディスク・ファーマ株式会社から購入した。 (4) Recombinant active factor VII (rFVIIa) solution Recombinant active factor VII (rFVIIa) (NovoSeven (registered trademark); Lot No. X1002) was purchased from Novo Nordisk Pharma Co., Ltd. did.
　実験の当日にｒＦＶＩＩａを蒸留水で溶解し、０．６ｍｇ／ｍＬ溶液を調製した。この０．６ｍｇ／ｍＬ溶液を生理食塩水で希釈し、１００００、３０００、及び１０００ｎｇ／ｍＬの濃度の溶液を調製し、氷中で保存して試験に用いた。
（実施例１）
ヒト（Ｐｏｏｌｅｄ）血漿を、３７℃の水浴で解凍し、それを氷中で保存して実験に使用した。
４５μＬの化合物（１）の溶液（１６．７及び３３．３μｇ／ｍＬ）を、４４５５μＬの血漿に添加した。４５μＬの５％ＤＭＳＯ－生理食塩水溶液を血漿に添加してコントロールとした。ＰＴは、血液凝固時間自動測定機（Ａｍｅｌｕｎｇ　ＫＣ－１０Ａ　ｍｉｃｒｏ，　Ｔｈｒｉｎｉｔｙ　Ｂｉｏｔｅｃｈ　Ｐｌｃ．）を用いて測定した。 On the day of the experiment, rFVIIa was dissolved in distilled water to prepare a 0.6 mg / mL solution. This 0.6 mg / mL solution was diluted with physiological saline to prepare solutions having a concentration of 10,000, 3000, and 1000 ng / mL, and stored in ice for use in the test.
Example 1
Human (Pooled) plasma was thawed in a 37 ° C. water bath and stored in ice for use in experiments.
45 μL of the solution of compound (1) (16.7 and 33.3 μg / mL) was added to 4455 μL of plasma. For control, 45 μL of 5% DMSO-saline solution was added to plasma. PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.).
　５μＬの乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］（上記の１５、５、及び１．５Ｕ／ｍＬの濃度の溶液）或いは生理食塩水と、４５μＬの化合物（１）の溶液又は５％ＤＭＳＯ－生理食塩水溶液を添加した血漿を、キュベットに加え、３７℃で１　分間プレインキュベーションした。 5 μL of dry human blood coagulation factor antibody bypass active complex [Feiba®] (solutions at concentrations of 15, 5, and 1.5 U / mL above) or saline and 45 μL of compound (1) Plasma added with solution or 5% DMSO-saline solution was added to the cuvette and preincubated for 1 minute at 37 ° C.
　混合液に、１００μＬのＨｅｍｏｓＩＬ　ＰＴ－ｆｉｂｒｉｎｏｇｅｎ　ＨＳ　ＰＬＵＳ　（Ｉｎｓｔｒｕｍｅｎｔａｔｉｏｎ　Ｌａｂｏｒａｔｏｒｙ，　Ｌｏｔ　Ｎｏ．　Ｎ０２９８４５８）を添加して凝固を開始させ、凝固時間を測定した。
（実施例２）
ヒト（Ｐｏｏｌｅｄ）血漿を、３７℃の水浴で解凍し、それを氷中で保存して実験に使用した。
４５μＬの化合物（１）の溶液（１６．７及び３３．３μｇ／ｍＬ）を、４４５５μＬの血漿に添加した。４５μＬの５％ＤＭＳＯ－生理食塩水溶液を血漿に添加してコントロールとした。
ＰＴは、血液凝固時間自動測定機（Ａｍｅｌｕｎｇ　ＫＣ－１０Ａ　ｍｉｃｒｏ，　Ｔｈｒｉｎｉｔｙ　Ｂｉｏｔｅｃｈ　Ｐｌｃ．）を用いて以下のように測定した。
５μＬの乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］（上記の１５、５、及び１．５Ｕ／ｍＬの濃度の溶液）、或いは生理食塩水と、４５μＬの化合物（１）の溶液又は５％ＤＭＳＯ－生理食塩水溶液を添加した血漿を、キュベットに加え、３７℃で１　分間プレインキュベーションした。
混合液に、１００μＬのＨｅｍｏｓＩＬ　ＰＴ－ｆｉｂｒｉｎｏｇｅｎ　ＨＳ　ＰＬＵＳ　（Ｉｎｓｔｒｕｍｅｎｔａｔｉｏｎ　Ｌａｂｏｒａｔｏｒｙ，　Ｌｏｔ　Ｎｏ．　Ｎ０２９８４５８）を添加して凝固を開始させ、凝固時間を測定した。
（実施例３）
ヒト（Ｐｏｏｌｅｄ）血漿を、３７℃の水浴で解凍し、それを氷中で保存した。
４５μＬの化合物（１）の溶液（１６．７及び３３．３μｇ／ｍＬ）を、４４５５μＬの血漿に添加した。４５μＬの５％ＤＭＳＯ－生理食塩水溶液を血漿に添加してコントロールとした。
ＰＴは、血液凝固時間自動測定機（Ａｍｅｌｕｎｇ　ＫＣ－１０Ａ　ｍｉｃｒｏ，　Ｔｈｒｉｎｉｔｙ　Ｂｉｏｔｅｃｈ　Ｐｌｃ．）を用いて以下のように測定した。
５μＬの遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）（上記の１００００、３０００、及び１０００ｎｇ／ｍＬの濃度の溶液）或いは生理食塩水と、４５μＬの化合物（１）又は５％ＤＭＳＯ－生理食塩水溶液を添加した血漿、又は５％ＤＭＳＯ－生理食塩水を、キュベットに加え、３７℃で１　分間プレインキュベーションした。
混合液に、１００μＬのＨｅｍｏｓＩＬ　ＰＴ－ｆｉｂｒｉｎｏｇｅｎ　ＨＳ　ＰＬＵＳ　（Ｉｎｓｔｒｕｍｅｎｔａｔｉｏｎ　Ｌａｂｏｒａｔｏｒｙ，Ｌｏｔ　Ｎｏ．Ｎ０２９８４５８）を添加して凝固を開始させ、凝固時間を測定した。
（試験結果）
実施例１の結果を図１にまとめた。実施例１において、コントロールである対照群のＰＴは１８．４±０．２秒であった（平均値±標準誤差、以下同様）。化合物（１）の１５０又は３００ｎｇ／ｍＬの濃度の添加により、ＰＴはそれぞれ３１．５±０．２秒又は４３．７±０．３秒に著しく延長した。乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］の単独投与（０．１５，０．５、及び１．５Ｕ／ｍＬ）は、濃度依存的にＰＴを短縮した。また、Ｆｅｉｂａ（登録商標）は、化合物（１）の添加によって引き起こされたＰＴの延長を、顕著にかつ濃度依存的に短縮した。Ｆｅｉｂａ（登録商標）の最高濃度（１．５Ｕ／ｍＬ）の投与では、１５０及び３００ｎｇ／ｍＬの化合物（１）の添加によってもたらされたＰＴ延長を、それぞれ１９．５±０．１秒及び２５．０±０．１秒に短縮した。 To the mixture, 100 μL of HemosIL PT-fibrinogen HS PLUS (Instrumentation Laboratory, Lot No. N0298458) was added to start coagulation, and the coagulation time was measured.
(Example 2)
Human (Pooled) plasma was thawed in a 37 ° C. water bath and stored in ice for use in experiments.
45 μL of the solution of compound (1) (16.7 and 33.3 μg / mL) was added to 4455 μL of plasma. For control, 45 μL of 5% DMSO-saline solution was added to plasma.
PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.) As follows.
5 μL of dried human blood coagulation factor IX complex [PPSB®-HT] (solutions at concentrations of 15, 5, and 1.5 U / mL above) or saline and 45 μL of compound (1 ) Or 5% DMSO-saline solution-added plasma was added to the cuvette and preincubated at 37 ° C. for 1 minute.
To the mixture, 100 μL of HemosIL PT-fibrinogen HS PLUS (Instrumentation Laboratory, Lot No. N0298458) was added to start coagulation, and the coagulation time was measured.
(Example 3)
Human (Pooled) plasma was thawed in a 37 ° C. water bath and stored in ice.
45 μL of the solution of compound (1) (16.7 and 33.3 μg / mL) was added to 4455 μL of plasma. For control, 45 μL of 5% DMSO-saline solution was added to plasma.
PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.) As follows.
5 μL of recombinant active factor VII (rFVIIa) (the solutions having the above concentrations of 10,000, 3000, and 1000 ng / mL) or physiological saline and 45 μL of compound (1) or 5% DMSO-physiological saline solution Added plasma or 5% DMSO-saline was added to the cuvette and preincubated for 1 minute at 37 ° C.
To the mixture, 100 μL of HemosIL PT-fibrinogen HS PLUS (Instrumentation Laboratory, Lot No. N0298458) was added to start coagulation, and the coagulation time was measured.
(Test results)
The results of Example 1 are summarized in FIG. In Example 1, the PT of the control group as a control was 18.4 ± 0.2 seconds (mean value ± standard error, the same applies hereinafter). Addition of 150 or 300 ng / mL concentration of compound (1) significantly extended PT to 31.5 ± 0.2 seconds or 43.7 ± 0.3 seconds, respectively. Single administration (0.15, 0.5, and 1.5 U / mL) of dry human blood coagulation factor antibody bypass activity complex [Feiba®] shortened PT in a concentration-dependent manner. In addition, Feiba® significantly shortened the PT prolongation caused by the addition of the compound (1) in a concentration-dependent manner. At the highest concentration of Feiba® (1.5 U / mL), the PT prolongation caused by the addition of 150 and 300 ng / mL of compound (1) is 19.5 ± 0.1 seconds and The time was shortened to 25.0 ± 0.1 seconds.
　また、実施例２の結果を図２にまとめた。実施例２において、コントロールである対照群のＰＴは、１７．８±０．０秒であった。化合物（１）の１５０又は３００ｎｇ／ｍＬの添加により、それぞれ３０．７±０．２秒又は４２．５±０．４秒に著しくＰＴが延長した。乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］の単独投与では、濃度依存的に（０．１５、０．５、及び１．５Ｕ／ｍＬ）、かつ著しくＰＴを短縮した。［ＰＰＳＢ（登録商標）－ＨＴ］（０．１５、０．５、及び１．５Ｕ／ｍＬ）は、化合物（１）の添加によるＰＴ延長を、顕著にかつ濃度依存的に短縮した。［ＰＰＳＢ（登録商標）－ＨＴ］の添加の最高濃度（１．５Ｕ／ｍＬ）では、１５０及び３００ｎｇ／ｍＬの化合物（１）の添加によるＰＴ延長を、それぞれ２３．７±０．１秒及び３１．６±０．４秒に短縮した。 The results of Example 2 are summarized in FIG. In Example 2, the PT of the control group as a control was 17.8 ± 0.0 seconds. The addition of 150 or 300 ng / mL of compound (1) significantly extended PT to 30.7 ± 0.2 seconds or 42.5 ± 0.4 seconds, respectively. Single administration of dry human blood coagulation factor IX complex [PPSB®-HT] concentration-dependently (0.15, 0.5, and 1.5 U / mL) and significantly shortened PT . [PPSB®-HT] (0.15, 0.5, and 1.5 U / mL) significantly shortened the PT prolongation due to the addition of Compound (1) in a concentration-dependent manner. At the highest concentration of addition of [PPSB®-HT] (1.5 U / mL), the PT extension with the addition of 150 and 300 ng / mL of compound (1) is 23.7 ± 0.1 seconds and The time was shortened to 31.6 ± 0.4 seconds.
　実施例３の結果を図３にまとめた。実施例３において、コントロールである対照群のＰＴは１７．９±０．２秒であった。化合物（１）の１５０又は３００ｎｇ／ｍＬの添加により、それぞれ３０．７±０．２秒又は４３．１±０．３秒にＰＴを著しく延長した。遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）の単独投与（１００、３００、及び１０００ｎｇ／ｍＬ）では、顕著にかつ濃度依存的にＰＴを短縮した。ｒＦＶＩＩａ（１００、３００、及び１０００ｎｇ／ｍＬ）の投与により、化合物（１）によって引き起こされたＰＴ延長を、顕著にかつ濃度依存的に短縮した。ｒＦＶＩＩａの最高濃度（１０００ｎｇ／ｍＬ）の投与では、は、１５０及び３００ｎｇ／ｍＬの化合物（１）の添加によるＰＴ延長を、それぞれ１７．３±０．２秒及び２４．２±０．３秒に短縮した。 The results of Example 3 are summarized in FIG. In Example 3, the PT of the control group as a control was 17.9 ± 0.2 seconds. The addition of 150 or 300 ng / mL of compound (1) significantly extended PT to 30.7 ± 0.2 seconds or 43.1 ± 0.3 seconds, respectively. PT administration was significantly and concentration-dependently shortened by the single administration of recombinant recombinant factor VII (rFVIIa) (100, 300, and 1000 ng / mL). Administration of rFVIIa (100, 300, and 1000 ng / mL) significantly shortened the PT prolongation caused by compound (1) in a concentration-dependent manner. For the administration of the highest concentration of rFVIIa (1000 ng / mL), the PT prolongation by the addition of 150 and 300 ng / mL of compound (1) is 17.3 ± 0.2 seconds and 24.2 ± 0.3 seconds, respectively. Shortened to
　実施例１～３の結果から、化合物（１）の添加によってもたらされたＰＴ延長に対する短縮効果は、遺伝子組換え活性型第ＶＩＩ因子（ｒＦＶＩＩａ）、乾燥人血液凝固因子抗体迂回活性複合体［Ｆｅｉｂａ（登録商標）］、及び乾燥人血液凝固第ＩＸ因子複合体［ＰＰＳＢ（登録商標）－ＨＴ］の順に高かった。
（比較例１）
　ヒト血漿に化合物（１）、血液凝固第ＶＩＩＩ因子製剤［コージネイト（登録商標）ＦＳ］を添加し、活性化部分トロンボプラスチン時間（ＡＰＴＴ）を血液凝固時間自動測定機（Ａｍｅｌｕｎｇ　ＫＣ－１０Ａ　ｍｉｃｒｏ，　Ｔｈｒｉｎｉｔｙ　Ｂｉｏｔｅｃｈ　Ｐｌｃ．）を用いて測定した。
を用いて測定した。血液凝固第ＶＩＩＩ因子製剤の臨床用量における血漿中濃度は約０．４４Ｕ／ｍＬと推定され［コージネイト（登録商標）ＦＳの添付文書］、今回の検討では０．００１?６．７Ｕ／ｍＬの濃度を用いた。 From the results of Examples 1 to 3, the shortening effect on PT prolongation caused by the addition of compound (1) indicates that recombinant active factor VII (rFVIIa), a dried human blood coagulation factor antibody bypass activity complex [ Feiba®] and dry human blood coagulation factor IX complex [PPSB®-HT] in that order.
(Comparative Example 1)
Compound (1) and a blood coagulation factor VIII preparation [Cognate® FS] are added to human plasma, and an activated partial thromboplastin time (APTT) is measured using a blood coagulation time automatic measuring machine (Amelung KC-10A micro, Trinity Biotech). Plc.).
It measured using. The plasma concentration of the blood coagulation factor VIII preparation at the clinical dose is estimated to be about 0.44 U / mL [package insert of Coordinate (R) FS]. In this study, the concentration was 0.001 to 6.7 U / mL Was used.
　その結果、ヒト血漿において、０．００１－１Ｕ／ｍＬの血液凝固第ＶＩＩＩ因子製剤単独ではＡＰＴＴに影響しなかったが、化合物（１）（１５０?４５０ｎｇ／ｍＬ）は濃度依存的にＡＰＴＴを延長した。化合物（１）３００ｎｇ／ｍＬによって１．９倍に延長したＡＰＴＴを血液凝固第ＶＩＩＩ因子製剤１Ｕ／ｍＬは１．６倍に短縮した。１Ｕ／ｍＬ以上の高濃度であるが、血液凝固第ＶＩＩＩ因子製剤６．７Ｕ／ｍＬでは、単独でＡＰＴＴをコントロール値の０．６倍に短縮し、化合物（１）による２倍延長をコントロール値レベルまで短縮した。
（比較例２）
　ヒト血漿に化合物（１）、血液凝固第ＩＸ因子製剤［クリスマシン（登録商標）－Ｍ］を添加し、活性化部分トロンボプラスチン時間（ＡＰＴＴ）を血液凝固時間自動測定機（Ａｍｅｌｕｎｇ　ＫＣ－１０Ａ　ｍｉｃｒｏ，　Ｔｈｒｉｎｉｔｙ　Ｂｉｏｔｅｃｈ　Ｐｌｃ．）を用いて測定した。クリスマシン（登録商標）－Ｍを５０Ｕ／ｋｇ（臨床用量付近）投与した時の血漿中濃度は約０．４７Ｕ／ｍＬと推定され［クリスマシン（登録商標）－Ｍ添付文書］、今回の検討では０．００１?６．７Ｕ／ｍＬの濃度を用いた。 As a result, in human plasma, the 0.001-1 U / mL blood coagulation factor VIII preparation alone did not affect APTT, but compound (1) (150-450 ng / mL) prolongs APTT in a concentration-dependent manner. did. Compound (1) APTT, which was extended 1.9 times by 300 ng / mL, was shortened to 1.6 times for blood coagulation factor VIII preparation 1 U / mL. Although the concentration is 1 U / mL or higher, the blood coagulation factor VIII preparation 6.7 U / mL alone shortens APTT to 0.6 times the control value and doubles the compound (1) extension to the control value. Reduced to level.
(Comparative Example 2)
Compound (1), blood coagulation factor IX preparation [Chris machine (registered trademark) -M] is added to human plasma, and activated partial thromboplastin time (APTT) is measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, (Thrinity Biotech Plc.). The plasma concentration is estimated to be about 0.47 U / mL when Chris Machine (registered trademark) -M is administered at 50 U / kg (near clinical dose) [Chris Machine (registered trademark) -M package insert]. In this case, a concentration of 0.001 to 6.7 U / mL was used.
　その結果、ヒト血漿において１Ｕ／ｍＬの血液凝固第ＩＸ因子製剤は単独でＡＰＴＴをコントロール値の０．８倍に短縮した。化合物（１）は、１５０?４５０ｎｇ／ｍＬで濃度依存的にＡＰＴＴを延長した。化合物（１）３００ｎｇ／ｍＬによって２倍に延長したＡＰＴＴを血液凝固第ＩＸ因子製剤１Ｕ／ｍＬは１．５倍に短縮した。検討可能な最高濃度（６．７Ｕ／ｍＬ）の血液凝固第ＩＸ因子製剤は、単独でＡＰＴＴをコントロール値の０．８倍に短縮し、化合物（１）による２倍延長を１．４倍まで短縮した。
（比較例３）
　一晩絶食したラットに０．５％メチルセルロースに懸濁した化合物（１）（３０ｍｇ／ｋｇ）を経口投与した。経口投与１５分後にラットをｔｈｉｏｐｅｎｔａｌｓｏｄｉｕｍ（１００ｍｇ／ｋｇ，ｉ．ｐ．）で麻酔した後、経口投与３０分後からラットの新鮮血漿（３０ｍＬ／ｋｇ／２ｈ：臨床用量の３倍の用量に相当）を頸静脈内に持続投与した。対照群には生理食塩液（３０ｍＬ／ｋｇ／２ｈ）を頸静脈内に持続投与した。新鮮血漿の持続静注の開始から２時間後に頸静脈からクエン酸加血液を採取した。採取した血液から血漿を分離し、ＰＴを血液凝固時間自動測定機（Ａｍｅｌｕｎｇ　ＫＣ－１０Ａ　ｍｉｃｒｏ，　Ｔｈｒｉｎｉｔｙ　Ｂｉｏｔｅｃｈ　Ｐｌｃ．）を用いて測定した。 As a result, the 1 U / mL blood coagulation factor IX preparation in human plasma alone shortened the APTT to 0.8 times the control value. Compound (1) prolonged APTT in a concentration-dependent manner at 150 to 450 ng / mL. Compound (1) APTT, which was doubled by 300 ng / mL, was shortened by 1.5 times for blood coagulation factor IX preparation 1 U / mL. The highest possible concentration (6.7 U / mL) of blood coagulation factor IX preparation alone shortens APTT to 0.8 times the control value and doubles the compound (1) to 1.4 times Shortened.
(Comparative Example 3)
Rats fasted overnight were orally administered compound (1) (30 mg / kg) suspended in 0.5% methylcellulose. Rats were anesthetized with thiopentasodium (100 mg / kg, ip) 15 minutes after oral administration, and then fresh rat plasma (30 mL / kg / 2h: equivalent to 3 times the clinical dose) 30 minutes after oral administration Was continuously administered into the jugular vein. In the control group, physiological saline (30 mL / kg / 2h) was continuously administered into the jugular vein. Citrated blood was collected from the jugular vein 2 hours after the start of continuous intravenous infusion of fresh plasma. Plasma was separated from the collected blood, and PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.).
　その結果、化合物（１）はＰＴを２．１４倍延長した。新鮮血漿(ＦＦ)は、臨床用量の３倍の高用量で、化合物（１）によるＰＴ延長を２．１４倍から１．６６倍に短縮した。
（比較例４）
　一晩絶食したラットに０．５％メチルセルロースに懸濁した化合物（１）（１０ｍｇ／ｋｇ）を経口投与し、その３０分後にビタミンＫ２（０．１及び１ｍｇ／ｋｇ）あるいは蒸留水を経口投与した。ビタミンＫ２投与の１時間４５分後にラットをｔｈｉｏｐｅｎｔａｌｓｏｄｉｕｍ（１００ｍｇ／ｋｇ, ｉ.ｐ.）で麻酔し、その１５分後に下大静脈からクエン酸加血液を採取した。採取した血液から血漿を分離し、ＰＴを血液凝固時間自動測定機（Ａｍｅｌｕｎｇ　ＫＣ－１０Ａ　ｍｉｃｒｏ，　Ｔｈｒｉｎｉｔｙ　Ｂｉｏｔｅｃｈ　Ｐｌｃ．）を用いて測定した。 As a result, Compound (1) extended PT by 2.14 times. Fresh plasma (FF) reduced PT prolongation by compound (1) from 2.14 times to 1.66 times at a dose 3 times higher than the clinical dose.
(Comparative Example 4)
Rats fasted overnight were orally administered compound (1) (10 mg / kg) suspended in 0.5% methylcellulose, and vitamin K2 (0.1 and 1 mg / kg) or distilled water was orally administered 30 minutes later. did. One hour and 45 minutes after vitamin K2 administration, the rats were anesthetized with thiopentasodium (100 mg / kg, ip), and citrated blood was collected from the inferior vena cava 15 minutes later. Plasma was separated from the collected blood, and PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Plc.).
　その結果、化合物（１）は経口投与２．５時間後にＰＴを１．３４倍延長した。ビタミンＫ２は、化合物（１）によって延長したＰＴに影響しなかった。
（比較例５）
　一晩絶食したラットをｔｈｉｏｐｅｎｔａｌ　ｓｏｄｉｕｍ（０．１ｇ／ｋｇ，ｉ．ｐ．）で麻酔した。麻酔１５分後から実験終了時まで、５％グルコース液あるいは化合物（１）を頸静脈より持続投与し（１ｍｇ／ｋｇ／ｈ）、投与開始２時間後に出血時間を測定した。出血時間測定の８分前に生理食塩液あるいはトラネキサム酸を大腿静脈より２分間かけて注入した。出血時間の測定は足底ｔｅｍｐｌａｔｅ法あるいは尾ｔｅｍｐｌａｔｅ法にて実施した。足底ｔｅｍｐｌａｔｅ法ではラットを仰臥位にし、右後肢足底にカッターナイフにて横一直線の長さ５ｍｍの切込みを入れた。３０秒毎に血液を濾紙に吸着させ、濾紙に血液が滲まなくなるまでの時間を最大３０分まで測定した。３０分以内に止血しなかった場合の出血時間は３０分とした。尾ｔｅｍｐｌａｔｅ法ではラットを仰臥位にし、尾先端より４ｃｍの位置の動脈部分に剃刀で深さ１ｍｍの切込みを入れた。１５秒毎に血液を濾紙に吸着させ、濾紙に血液が滲まなくなるまでの時間を最大３０分まで測定した。なお、出血時間測定開始の３分前に頸静脈より１／１０容の３．１３％　ｔｒｉｓｏｄｉｕｍ　ｃｉｔｒａｔｅ　ｄｉｈｙｄｒａｔｅ加血液０．５ｍＬを採血した。採取した血液から血漿を分離後（１，５００×ｇ、１０ｍｉｎ、４℃）、ＰＴを測定した。 As a result, Compound (1) prolonged PT by 1.34 times 2.5 hours after oral administration. Vitamin K2 did not affect PT prolonged by compound (1).
(Comparative Example 5)
Rats fasted overnight were anesthetized with thiopental sodium (0.1 g / kg, ip). From 15 minutes after anesthesia to the end of the experiment, 5% glucose solution or compound (1) was continuously administered from the jugular vein (1 mg / kg / h), and the bleeding time was measured 2 hours after the start of administration. Saline solution or tranexamic acid was injected over 2 minutes from the femoral vein 8 minutes before the bleeding time measurement. The bleeding time was measured by the plantar template method or the tail template method. In the plantar template method, the rat was placed in a supine position, and a 5 mm long incision was made with a cutter knife in the sole of the right hind limb. The blood was adsorbed on the filter paper every 30 seconds, and the time until blood did not bleed on the filter paper was measured up to 30 minutes. If the hemostasis was not stopped within 30 minutes, the bleeding time was 30 minutes. In the tail template method, the rat was placed in a supine position, and a 1 mm deep incision was made with a razor into the arterial portion 4 cm from the tip of the tail. The blood was adsorbed on the filter paper every 15 seconds, and the time until blood did not bleed on the filter paper was measured up to 30 minutes. In addition, 3 minutes before the start of measurement of bleeding time, 0.5 mL of 1/10 volume of 3.13% trisodium citrate dihydrate was collected from the jugular vein. After separating plasma from the collected blood (1,500 × g, 10 min, 4 ° C.), PT was measured.
　その結果、化合物（１）の投与により足底出血時間が有意に延長した。この出血時間の延長に対し、３０、１００及び３００ｍｇ／ｋｇのトラネキサム酸の投与では抑制作用を示さなかった。化合物（１）によってＰＴが１．３６倍延長し、３０ｍｇ／ｋｇのトラネキサム酸併用群でも同等のＰＴの延長が認められた。また、化合物（１）の投与によりラット尾出血時間が有意に延長した（延長率は１．７７倍）。この出血時間の延長に対し、３０及び１００ｍｇ／ｋｇのトラネキサム酸の投与では抑制作用を示さなかった。化合物（１）によってＰＴが１．３３倍延長し、トラネキサム酸併用群でも同様にＰＴが延長し、化合物（１）の投与によるＰＴ延長の抑制効果は見られなかった。 As a result, plantar bleeding time was significantly prolonged by administration of compound (1). In contrast to this prolonged bleeding time, administration of 30, 100 and 300 mg / kg tranexamic acid showed no inhibitory effect. With compound (1), PT was extended 1.36 times, and the same PT extension was observed in the 30 mg / kg tranexamic acid combination group. In addition, administration of compound (1) significantly prolonged rat tail bleeding time (the rate of extension was 1.77 times). In contrast to this prolonged bleeding time, administration of 30 and 100 mg / kg tranexamic acid showed no inhibitory effect. The compound (1) prolongs PT by 1.33 times, and the PT in the tranexamic acid combination group similarly prolongs PT, and no inhibitory effect on PT prolongation by administration of compound (1) was observed.
　さらに、Ｃｌｏｔ溶解時間測定系にｃｏｎｔｒｏｌ血漿を添加すると、ｃｌｏｔ形成後（吸光度上昇）にｃｌｏｔ溶解が急速に生じ（吸光度低下）、その溶解時間は約４０?５０分であった。３０及び１００ｍｇ／ｋｇのトラネキサム酸投与血漿では、１８０分までｃｌｏｔ溶解が認められなかったことから、トラネキサム酸の投与により、その主作用である線溶系抑制が確認された。 Furthermore, when control plasma was added to the Clot dissolution time measurement system, clot dissolution occurred rapidly (absorbance decrease) after clot formation (absorbance increase), and the dissolution time was about 40-50 minutes. In 30 and 100 mg / kg tranexamic acid-administered plasma, clot dissolution was not observed until 180 minutes, and thus the administration of tranexamic acid confirmed the suppression of the fibrinolytic system, which is the main action.
　以上から、ラット足底及び尾出血時間モデルを用いて、化合物（１）による出血時間延長に対し、トラネキサム酸は３０、１００及び３００ｍｇ／ｋｇの用量において化合物（１）の作用のリバース作用を示さなかった。 From the above, tranexamic acid showed a reverse action of compound (1) at doses of 30, 100, and 300 mg / kg, with respect to prolonged bleeding time by compound (1) using rat plantar and tail bleeding time models. There wasn't.
　本願発明は、化合物（１）、薬理上許容される塩、又はそれらの水和物が、例えば過剰投与等によって万が一に出血事故が発生した場合、薬物の抗凝固作用をリバースするリバース剤及び／又はリバース方法として利用できる。 The present invention relates to a reverse agent that reverses the anticoagulant action of a drug in the event that a bleeding accident occurs due to, for example, excessive administration of compound (1), a pharmacologically acceptable salt, or a hydrate thereof. Alternatively, it can be used as a reverse method.

Claims (4)

  1. 　ＦＸａ阻害薬の抗凝固作用のリバース剤であって、
    乾燥人血液凝固因子抗体迂回活性複合体、乾燥人血液凝固第ＩＸ因子複合体、及び、遺伝子組換え活性型第ＶＩＩ因子からなる群より選ばれる１又は２以上であるリバース剤。     A reverse agent of the anticoagulant action of an FXa inhibitor,
    A reverse agent that is one or more selected from the group consisting of a dried human blood coagulation factor antibody bypass activity complex, a dried human blood coagulation factor IX complex, and a recombinant recombinant factor VII.
  2. 　ＦＸａ阻害薬が、下記式（１）
    Figure JPOXMLDOC01-appb-C000001
    で表されるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド、薬理上許容される塩、又はそれらの水和物である、請求項1に記載のリバース剤。     FXa inhibitor is represented by the following formula (1)
    Figure JPOXMLDOC01-appb-C000001
    N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4 , 5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, a pharmacologically acceptable salt, or a hydrate thereof. The reverse agent described in 1.
  3. 　ＦＸａ阻害薬が、下記式（１ａ）
    Figure JPOXMLDOC01-appb-C000002
    で表されるＮ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド・ｐ－トルエンスルホン酸塩・１水和物である、請求項１に記載のリバース剤。     FXa inhibitor is represented by the following formula (1a)
    Figure JPOXMLDOC01-appb-C000002
    N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4 , 5,6,7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide / p-toluenesulfonate / monohydrate Reverse agent.
  4. 　乾燥人血液凝固因子抗体迂回活性複合体、乾燥人血液凝固第ＩＸ因子複合体、及び、遺伝子組換え活性型第ＶＩＩ因子からなる群より選ばれる１又は２以上を用いることを特徴とする、Ｎ１－（５－クロロピリジン－２－イル）－Ｎ２－（（１Ｓ，２Ｒ，４Ｓ）－４－［（ジメチルアミノ）カルボニル］－２－｛［（５－メチル－４，５，６，７－テトラヒドロチアゾロ［５，４－ｃ］ピリジン－２－イル）カルボニル］アミノ｝シクロヘキシル）エタンジアミド、薬理上許容される塩、又はそれらの水和物のＦＸａ阻害作用をリバースする方法。 1 or 2 or more selected from the group consisting of a dried human blood coagulation factor antibody bypass activity complex, a dried human blood coagulation factor IX complex, and a genetically modified active factor VII, N 1- (5-chloropyridin-2-yl) -N 2 -((1S, 2R, 4S) -4-[(dimethylamino) carbonyl] -2-{[(5-methyl-4,5,6, 7. A method for reversing the FXa inhibitory action of 7-tetrahydrothiazolo [5,4-c] pyridin-2-yl) carbonyl] amino} cyclohexyl) ethanediamide, a pharmacologically acceptable salt, or a hydrate thereof.
PCT/JP2011/072106 2010-09-28 2011-09-27 Reversing agent for anticoagulants WO2012043577A1 (en)

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