TW201219404A - Reversal agents for the anticoagulant effect - Google Patents

Abstract

The object of present invention is to provide a reversal agent capable of reversing the anticoagulant effect of FXa inhibitor; and a reversal method by using the same. The solution is a reversal agent, which is a reversal agent of the anticoagulant effect of oral anticoagulant, and is selected from one or two or more of group consisting of dry human blood coagulation factor antibody roundabout activity complex, dry human blood coagulation factor IX complex, and gene recombinant activity type factor VII.

Description

201219404 VI. Description of the invention: [Technical field to which the invention belongs]

The present invention relates to a reversal agent which reverses the anticoagulant effect of an activated blood coagulation factor X inhibitor; and/or a reversal method using a reversal agent. [Prior Art] Heparin and warfarin are known as anti-coagulant drugs for inhibiting blood coagulation. [In addition, as a new mechanism of action, anticoagulants are currently developing an activated blood coagulation. The factor X inhibitor (hereinafter also referred to as a FXa inhibitor) is directly inhibited by the activated blood coagulation factor X (hereinafter also referred to as FXa). These anticoagulants are known to have a blood coagulation inhibitory effect based on their action mechanism, and side effects are a risk of bleeding. Therefore, when an anticoagulant is administered excessively, or when a major bleeding occurs, it is necessary to have a drug or a method for reversing (neutralizing) the blood coagulation suppressing action of the drug to return the normal coagulation time to a normal value. As an anticoagulant neutralizing agent or a reversing agent for anticoagulants, for example, protamine (protamine sulfate) for heparin and vitamin K1 for warfarin are known (refer to, for example, non-patent literature).丨~2). Further, it is known that non-specific hemostatic agents are known as fresh frozen plasma (FFP), and the neutralizing agent or neutralization method which is applicable and most suitable according to each anticoagulant is also different. As such, the neutralizing agent that acts will vary depending on the mechanism of action of the anticoagulant, thus providing the most suitable neutralizing agent or neutralizing method for drugs having various anticoagulant effects, in thrombosis and embolism The treatment side 201219404 is an important treatment in the case of excess dose of medication or major bleeding. Ν匕(5-gas η than pyridin-2-yl s 2R 4S)-4-[(didecylamino)carbonyl]_2_{[(5·曱基_4, represented by the following formula (1), 5,6,7_tetrahydrogen sold. Sodium [5,4-c]D-pyridin-2-yl)carbonyl]amino}cyclohexyl)ethanediamine (hereinafter also abbreviated as compound (1) ) situation]

Pharmacologically acceptable salts, or such hydrates, have a strong direct inhibitory effect on FXa's clinically proven to be effective as a preventive for blood tests and embolisms, especially deep vein thrombosis (Deep Vein thrombosis : DVT), Venous Thromboembolism (VTE), and blood tests based on non-valvular Atrial Fibrillatiorl (NVAF). Prophylactic and therapeutic drugs for embolism, atrium A stroke preventive medicine for patients with tremors (see, for example, Patent Documents 1 to 2 and Non-Patent Documents 3 to 5). Furthermore, the literature also reports low-molecular FXa inhibitors, in addition to the above-mentioned anticoagulant thrombosis and embolism such as deep vein thrombosis, venous thrombosis and embolism (VTE), and non-valvular atrial fibrillation (NVAF). In addition to the prevention of stroke, embolism and atrial fibrillation in patients with cerebral infarction, cerebral infarction, cerebral embolism, myocardial infarction, angina, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS) Berger's disease, pan--4-201219404 intravascular coagulation syndrome, thrombosis after replacement of valvular/articular joints, thrombosis after revascularization, and reocclusion 'MODS insufficiency (MODS), in vitro Effects such as thrombus formation during circulation or blood coagulation at the time of blood collection (see, for example, non-patent literature) "Exhibition of compound (1), pharmacologically acceptable salt, or hydrate of these hydrates in clinical tests] The risks of prevention and treatment under the dosage of administration are small. However, for example, due to excessive administration of compound #(1), pharmacologically acceptable salt, or such hydrates, etc. A reversal agent and/or a reversal method for reversing the anticoagulant effect of a drug in the event of a bleeding accident is still unknown. [Prior Art Literature] [Patent Literature] [Patent Document 1] International Publication No. 2003/000680 pamphlet [Patent Literature 2] International Publication No. 20〇4/〇5 87 1 5 pamphlet [Non-patent literature] [Non-Patent Document 1] Pharmaceutical Supplementary Document Heparin Antagonist 曰 Pharmacopoeia Protamine Protease Injection 20 1 July Revised (1st edition) [Non-Patent Document 2] Supplementary Description of Pharmaceuticals Vitamin K 1 Preparations 2 July 2010 Revision (Revision 2nd Edition) [Non-Patent Document 3] JI Weitz et al., Thrombosis and

Haemostasis, 2010, 1 04(3), 633~64 1 [Non-Patent Document 4] G. Raskob et al., Thrombosis and

Haemostasis, 201 0, 1 04(3), 642~649 [Non-Patent Document 5] T. Furugohri et al., Journal of

Thrombosis and Haemostasis, 6; 1 542~1 549, 2008 201219404 [Non-Patent Document 6] Robert J. Leadley, Jr., Current

Topics in Medicinal Chemistry, 2001. 1,151 to 159 [Problems to be Solved by the Invention] An object of the present invention is to provide a reversal agent which is pharmaceutically pharmaceutically acceptable in excess of compound (1). Salt, or such hydrates, or when a major outbreak occurs, 'reversal of prolonged blood coagulation time; and/or reversal method>> [Means for solving the problem] The present inventors have for the compound (1), pharmacologically Allowable salts, or such hydrates, which are agents that neutralize the anticoagulation effect, are used in the in vitro test (in vitr〇) using prothrombin time. (PT), etc., as a target, a dried human blood coagulation factor-factor complex [PPSB (registered trademark) ht] used as a therapeutic agent for hemophilia or a therapeutic agent for blood coagulation factor IX deficiency, and dried After the human blood coagulation factor antibody bypass active complex 2; (registered trademark)], and the recombinant active type νη factor coagulation = the degree of agronomy depends on the blood-interstitial elongation extended by the compound 1 carry out this invention. That is, the present invention provides the following (1) to (4). (1 a): a neutralizing agent, such as / coagulation anticoagulant coagulation effect and agent, selected from the human blood path active pleats after drying # liquid coagulation factor antibody side modification & The body's blood coagulation after drying, and the base of the tongue. The work &, the factor factor complex is due to the recombinant active factor VII factor in the box. (lb): a neutralizing agent, which is a 201219404 agent in the coagulation of FXa 1 Tetra Pak, which is selected from the group consisting of dried human blood coagulation factor antibody bypassing abruptly complex β-body. One or more of the human ΑΒΕ1 tongue 4 1 衣 固 固 IX factor complex and the gene recombinant active type V „ factor after drying. (1)· A reversal agent, which will be a 吴 ^ Wu FXa inhibitor The reversal agent for coagulation is selected from the group consisting of dry de & λ # & ', followed by human blood sputum factor antibody bypass activity ^ human dry blood coagulation factor IX complex, and recombination One or more of the active factor VII. The neutralizing agent described in (lb), wherein N _(5-chloropyridin-2-yl) represented by i _ ) in the FXa inhibitory drug system )-N2-((l S,2R,4S)-4-amino)|ί炭基]_2-{[(5·曱基_4,5,6,7·tetrahydrothiazole 5.4 - r 1 Oll

(2) The reversal agent according to (1), wherein the FXa inhibitor is N丨_(5_gas acridine_2_ylamine-monomethylamino)carbonyl represented by the following formula (1) ]-2-{[(5-Methyl-4,5,6,7-tetrahydrothiazolo[5'4-C]°pyridin-2-yl)carbonyl]amino}cyclohexyl)ethane II Guanamine:

C1 (1) 201219404, pharmacologically acceptable salt, or such hydrate. (3a): a neutralizing agent as described in (ib), wherein the FXa inhibitory drug system is represented by the formula (la): n1-. -chloropyridin-2-yl)-N2-((1S,2R,4S)-4·[(dimethylamino)carbonyl]·2·{[(5_methyl_4,5,6,7 · tetrahydrothiazepine [5,4-c]pyridin-2-yl)carbonyl]amino}cyclohexyl)ethanediamine ρ-甲

(3) The reversal agent according to (1), wherein the FXa inhibitor is Νι_(5_pyridin-2-yl)-N2-((lS, 2R, 4S) represented by the following formula (la) -4-[(dimethylamino)carbonyl]-2-{[(5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]°pyridin-2-yl )carbonyl]amino}cyclohexyl)ethanediamine.

(4a): - Kind will be N1-. -chloropyridin-2-yl)-methylamino)amino]mercapto-4,5,6,7-tetrahydroanthracene and [5'4 c]tbdi-2-yl)carbonyl]amino a cyclohexyl)diamine, a pharmacologically acceptable salt, or a method for neutralizing FXa inhibition of such hydrates, characterized by the use of a human blood coagulation factor anti-201219404 body bypass selected from the group after drying The active complex, the dried human blood coagulation first χ factor complex, and the recombinant recombinant active νΠ factor are more than the above species. (4) · One will be Ν1-. -chloropyridin-2-yl)-N2-((lS,2R,4S)-4-[(dimethylamino)-yl]-2-{[(5-methyl·4,5,6 , 7-tetrahydrothiazolo[5,4-c]acridin-2-yl)carbonyl]amino}cyclohexyl)ethanedioxime, a pharmacologically potable salt, or a hydrate thereof The method for reversing the inhibition of FXa is characterized by using a human blood coagulation factor antibody bypass active complex selected after drying, a human blood coagulation factor X complex after drying, and a recombinant active type νπ factor. More than one species. [Effect of the invention; | ' can provide a reversal agent which is based on the thrombus-plug FXa inhibitor νΌ-α-pyridine-2-yl)·Ν methylamino) benzyl]_2_{[(5-methyl,

[Embodiment] In the treatment, an anticoagulant or the like is administered by mistake or excessively, for example, -((lS, 2R, 4S)-4-[(di-methyl-4, 5, 6, 7) - tetrahydrothiazolo[5,4-c] hexyl)diacetamide, pharmacologically acceptable to cause an increase in the risk of bleeding, or to reverse the anticoagulant effect of the drug; and / or [to implement Form of invention]

A specific example of the direct-acting type I fAa inhibitor represented by the formula (1) is a preferred example of the above-mentioned compound [hereinafter, abbreviated as the case of the compound (1)] 201219404 is preferable. The compound (1) may be a free form (free base) or a hydrate thereof, and may be a pharmacologically acceptable salt or a salt hydrate. The pharmacologically acceptable salt of the compound represented by the formula (1) may, for example, be a hydrochloride, a sulfate, a hydrobromide, a hydrocyanate, a phosphate, a nitrate, a benzoate or an oxime sulfonate. , 2-hydroxyethane sulfonate, p-toluenesulfonate, acetate, propionate, oxalate, malonate, succinate, sebacate, adipate, tartrate, horse Acidate, fumarate, malate, mandelic acid, etc. The salt of the compound represented by the formula (1) is preferably a hydrochloride or a p-toluenesulfonate; and a p-toluenesulfonate is particularly preferred. For the compound represented by the following formula (1),

Suitable materials can be enumerated as follows: Heart-(5-chloropyridin-2-yl)-N2-((lS,2R,4S)-4-[(didecylamino)carbonyl]-2-{[( 5-methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]acridin-2-yl)carbonyl]amino}cyclohexyl)ethanediamine; Benzene-2-yl)-N2-((lS,2R,4S)-4-[(dimethylamino)carbonyl]-2-{[(5-methyl-4,5,6, 7-tetrahydrothiazolo[5,4-Cp-pyridin-2-yl)carbonyl]amino}cyclohexyl)ethanedioxanamide hydrochloride; -(5-gas.pyridin-2-yl) )-N2-((lS,2R,4S)-4-[(dimethylamino)carbonyl]-2-{[(5-mercapto-4,5,6,7-tetrahydrothiazolo[5 4-〇]pyridin-2-yl)carbonyl]amino}cyclohexyl)ethanedioxime•p-toluenesulfonate; and-10-201219404 N1-. -chloropyridin-2-yl)-N2-((lS,2R,4S)-4-[(dimethylamino)carbonyl]-2-{[(5-methyl-4,5,6,7 - Tetrathiathiazolo[5,4-c]pyridin-2-yl)carbonyl]amino}cyclohexyl) phenyl diamine, p-benzoic acid hydrochloride. Among the above-mentioned suitable compounds, <^-(5-gas "bipyridin-2-yl)_n2-((1S,2R,4S)-4-[(2) represented by the following formula (la) Methylamino)carbonyl]-2-{[(5-methyl- 4,5,6,7-tetrahydro" stopper. Sit and [5,4-c]0 ratio. Alkylamino}cyclohexyl)ethanediamine-p-sulfonate·1 hydrate [hereinafter, abbreviated as the case of compound (丨a)] is particularly good.

And a compound which removes "acid" and "hydrate" of the hydrate, for example, with the above-mentioned salt (acid addition salt) and/or the acid which forms a hydrate. For example, the free base (free state) of the above-mentioned octahydrate (1 a) means hydrazine represented by the following formula (1). 〇 〇 · 2 2 2 2 2 2 2 2 2 4^]pyridinyl]amino)cyclohexyl)ethanediamine (1): soil)

-11 - 201219404 The amount of the compound (1) represented by the formula (1) in the present specification is the amount of the compound (1) which is converted into the free base (free form). N1-(5-Gas ratio bit-2-yl)-N2-((l S,2R, 4S)-4-[(didecylamino)carbonyl]_2_{[(5曱) as FXa inhibitor _4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)carbonyl]aminopurine cyclohexyl)ethanedioxime·p-toluene 4 salt·1 hydrate The amount of (Compound la) to be administered is preferably 5 mg to 9 mg based on the free base (free form) [the above compound (1)]; and preferably 15 mg to 60 mg. In the case of administration as an oral blood coagulation inhibitor, -(5_chloro.by bite_2_yl)_Ν2_((ΐδ 2ιι, 4^4 [(dimethylamino) Carbonyl]-2-{[(5-methyl·4,5,6,7-tetrahydrothiazolo[5 4_c]pyridin-2-yl)carbonyl]amino}cyclohexyl)ethanediamine, The salt or the hydrates thereof are administered to the patient together with a pharmacologically acceptable additive in the form of a dosage form such as a tablet, etc., which is administered as a crypto-inhibiting agent, N1]5-chloro 》N2_((1S,2R,4S)_4 heptamethylaminomethyl]-2·{[(5-methyl-4,5,6,7_tetrahydro 0 〇 〇 and [5,4_^ is more than di-2-yl)carbonyl]amino}cyclohexyl)ethanediamine as the free base (the content of the free 亡 亡 化合物 化合物 ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( Good, 15mg, 30mg and 60mg are particularly good. ...... 义付削" means the agent that neutralizes or reverses the anticoagulant effect of anticoagulant. The neutralization or reversal of action is also interpreted as Detoxification is caused by the use of drugs and the undesirable side effects of the human body. The "neutralizer" or "reversal agent" of the drug can be listed as the human blood coagulation factor antibody bypass active complex [Feiba (registered trademark)], dried after -12-201219404 Human blood coagulation factor IX complex [PPSB (registered trademark)_HT], recombinant recombinant factor VII factor (rFVna), blood coagulation factor VIII preparation [Kogenate (registered trademark) FS], blood coagulation π factor preparation [Christmassin (registered trademark, fresh frozen plasma (FFP: Fresh Frozen Plasma), vitamin κ (including vitamin κι, vitamin K2), tranexamic acid 荨. Here 'dry human gold solution coagulation factor IX complex [PPSB (registered trademark)·ΗΤ], means that it is contained in the Interview Form [April 2009 (3rd edition)] of PPSB (registered trademark) ΗΤ "NICHIYAKU" a blood coagulation factor IX preparation (20 units/mL or more), and in addition to blood coagulation factor IX, other vitamin K-dependent blood coagulation factors of factor II, VII and X "thrombin" Original complex concentrate preparation (PCC: Prothrombin Complex Concentrate). The "reversal agent" or "neutralizer" in the application of the present invention is a human blood coagulation factor antibody bypass active complex [Feiba (registered trademark)] after drying. Drying into blood coagulation factor IX complex [PPSB. (registered trademark)-HT], and gene recombinant active form νπ factor (rFVIIa) is preferred; dried human liquid coagulation factor antibody bypass active complex [Feiba (registered trademark)] and gene recombinant active factor VII (rFvna) are preferred. The reversal effect of the anticoagulant effect in the specification of the present application was evaluated by using the zymogen time (pT) and the activated partial thrombin time (Αρττ) as indicators. Further, the coagulation time was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Trinity Biotech Pic.). • 13- 201219404 The statistically significant analysis of the data in this specification uses the Dunnett method. P# is indicated by ###, p<0.001 is represented by $$, and 0.001 is relative to the control group (the compound (1) and the reversal agent are absent). A significant difference of P &lt; 0_001 relative to the compound (1) group is indicated by ***. The thrombus and embolism in the present specification may, for example, be as follows: selected from the group consisting of "cerebral infarction, cerebral embolism, myocardial infarction, angina, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS), Berger's disease) Deep venous thrombosis, generalized intravascular coagulation syndrome, thrombosis after prosthetic valve/joint replacement, gold plug formation and reocclusion after revascularization, multiple organ insufficiency (MODS), thrombosis during cardiopulmonary bypass, or One or more diseases in the group consisting of blood coagulation, venous thrombosis, embolism (VTE), and non-valvular atrial fibrillation (NVAF) at the time of blood collection, or thrombosis and embolism caused by such diseases, however Completely free from these diseases. ▲•栓. Embolism is caused by cerebral infarction: cerebral congestion, myocardial infarction, angina, pulmonary infarction, pulmonary embolism, acute coronary syndrome (ACS), Berger's disease, deep venous thrombosis, generalized Blood test formation after coagulation syndrome, artificial valve/Guanlang replacement, thrombosis and reocclusion after hematogenous reconstruction, multiple organ insufficiency (mods), thrombosis during cardiopulmonary bypass, blood coagulation during blood collection, venous thrombosis · Embolism (VTE), as well as thrombosis and embolism based on non-valvular atrial fibrillation (NVAF); venous thrombosis, embolism (VTE), and non-valvular atrial fibrillation (NVAF) Thrombosis and embolism are preferred. The formulation in the present specification means an oral preparation, and the active ingredient and the additive of the music science are together with a dosage form such as a suspect, a granule 14 - 201219404, a powder, a capsule or a syrup. Inject to the patient. The pharmacologically acceptable additives may, for example, be excipients, disintegrating agents, binding agents, flow agents, lubricants, colorants, and gloss. [Embodiment] The present invention will be described in detail below by way of examples and the present invention is completely un These embodiments are defined. [Preparation of the test solution for evaluation] (1) The solution of the compound (1) was weighed and diluted with 5% DMS 〇 _ physiological saline solution on the test day, and the concentration was adjusted to 33.3 μέ/ηα, and 167^ A solution of /mL [concentration of compound (1) converted to the free base] was stored under light-shielding conditions and room temperature for testing. (2) Human gold coagulation factor antibody bypass active complex after drying of human blood coagulation factor antibody bypass active complex [Feiba (registered trademark)] solution [Feiba (registered trademark)] (L〇t No . VNF1H040 ; active ingredient: 500U) purchased from Baxter Co., Ltd. On the day of the experiment, Feiba (registered trademark) was dissolved in distilled water to prepare a 50 U/mL solution. The 50 U/mL solution was diluted with physiological saline to prepare solutions having concentrations of 15, 5, and 1.5 U/mL, and stored in ice for testing. U) Human blood coagulation factor IX complex after drying [PPSB (registered trademark)-HT] solution dried human blood coagulation factor IX complex [PPSB (registered trademark)-HT] (Lot No. N117DK; 200U ) purchased from Japan Pharmaceutical Co., Ltd. 201219404 Limited company β On the day of the experiment, [PPSB (registered trademark)-ΗΤ] was dissolved in distilled water to prepare a 20 U/mL solution. This 2 U/mL solution was diluted with physiological saline to prepare a solution having a concentration of 15, 5, and 15 U/niL, and stored in ice for testing. (4) Recombinant active factor VII (rFVila) solution Recombinant active factor VII (rFVIIa) (NovoSeven (registered trademark); Lot No. XI 〇〇 2) was purchased from Novo Nordisk Pharma Co., Ltd. On the day of the experiment, rFVIIa was dissolved in distilled water to prepare a solution of 〇.6 mg/mL. This 〇 6 mg/mL solution was diluted with physiological saline to prepare a solution having a concentration of 10,000, 3,000, and 10 ng/mL, and stored in ice for testing. (Example 1) Human (Pooled) plasma was thawed in a 37 ° C water bath and stored in ice for experimental use. A solution of 45 41^ of the compound (1) (167 and 33 3 /1111^) was added to 4455 41^ of plasma. 454 [5% of 01^0-physiological saline solution was added to the plasma as a control group. The PT system is measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Thrinity Biotech Pic.). ο 5 μί of dried human blood coagulation factor antibody bypass active compound 雔 [Feiba (i registered trademark) (The above concentrations of 15, 5, and 1.5 U / mL solution) or physiological saline, 45 吣 of the compound (1) solution or 5% DMSO - physiological saline solution of the plasma was added to the colorimetric tube, at 37 - 16- 201219404 Pre-treatment for 1 minute. To the mixture, 1 〇〇 pL of H emosIL 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 experimental use. A solution of 45 41^ of the compound (1) (16.7 and 33.3 #8/1111^) was added to 445 5 pL of plasma. 45 μL of a 5% DMSO-physiological saline solution was added to the plasma as a control group. The PT was measured using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Thrinity Biotech Pic.) as follows. 5 μί of dried human blood coagulation factor IX complex [PPSB (registered trademark)_ΗΤ] (solutions of the above concentrations of 15, 5, and 1.5 U/mL), or physiological saline, 45 pL of compound The solution of (1) or the plasma of 5% DMSO-physiological saline solution was added to a colorimetric tube and pretreated at 37 ° C for 1 minute. HemosIL PT-fibrinogen HS PLUS (Instrumentation Laboratory, Lot No. N0298458) of ΙΟΟμί was added to the mixture 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 pL of the compound (1) solution (16.7 and 33.3 pg/mL) was added to the plasma of -17-201219404 to 4455 pL. 45 pL of a 5% DMSO-physiological saline solution was added to the plasma to determine a control group. PT is measured by the following method using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Thrinity Biotech Pic.). 5 pL of recombinant active type factor VII (rFVIIa) is added (the above concentrations are 10,000, 3000, And 1000 ng / mL of the solution) or physiological saline, 45 pL of the compound (1) or 5% DMSO - physiological saline solution of plasma, or 5% DMSO - physiological saline added to the colorimetric tube 'pretreatment at 37 ° c minute.

100 μL of HemosIL PT-fibrinogen HS PLUS (Instrumentation Laboratory, Lot No. N0298458) was added to the mixture to start coagulation, and the coagulation time was measured. (Test Results) The results of Example 1 were summarized in Fig. 1. In Example 1, the PT of the control group as a control group was 18.4 ± 0.2 hours (the average value is equal to or less than the standard error). By adding a compound (1) at a concentration of 150 or 300 ng/mL, PT was significantly prolonged to 31.5 ± 0.2 seconds or 43. 7 ± 〇 · 3 seconds, respectively, and administered alone (0.15, 0.5, dried human blood) Coagulation factor antibody bypass active complex [Feiba (registered trademark gate, the ρτ concentration is shortened depending on. In addition, Feiba (registered trademark) will significantly increase the PT caused by the addition of compound (1) and concentration dependently Shortening. When Feiba (registered trademark) was administered at the highest concentration (1.5 U/mL), the ρτ extension by the addition of 150 and 300 ng/mL of compound (1) was shortened to 19.5±0·1 second, respectively. And 25.0±0.1 sec. -18 - 201219404 In addition, the results of Example 2 were summarized in Fig. 2. In Example 2, the PT of the control group as a control group was 丨7 8d: leap seconds. Addition of 150 or 300 ng/mL of compound (1) significantly prolongs ρτ to 3 0.7 ± 0.2 sec or 42.5 ± 0.4 sec. Human blood coagulation factor IX complex after drying alone [PPSB (registered trademark) When _ht], the concentration of Ρτ is dependent (0.15, 0.5, and 1.5 U/mL) and is significantly shortened. [ppSB (registered trademark)-HT] (0.15, 0.5, and 1.5 U/mL) significantly prolonged the PT produced by the addition of the compound (1) and decreased in a concentration-dependent manner. It was added at the highest concentration (l'5 U/mL) [ PPSB (registered trademark)_HT] shortened the ρτ elongation by the addition of 150 and 300 ng/mL of compound (1) to 23.7 ± 0.1 sec and 31.6 ± 0.4 sec, respectively. The results of Example 3 were summarized in the first In the third embodiment, the ruthenium of the control group as a control group was 17.9±〇.2 sec. By adding 15 〇 or 300 ng/mL of the compound (1) ' respectively, the Ρτ was significantly prolonged to 3 分别. 7±〇2 or 43·1±0.3 sec. When the recombinant recombinant active γn factor (rFVIIa) (100, 300 and 1000 ng/mL) was administered alone, the ρΤ was significantly reduced and the concentration was shortened. Administration of rFVIIa (10, 300, and i〇00 ng/mL) resulted in a significant prolongation of sputum caused by the compound (1) and a concentration-dependent shortening. When rFVIIa was administered at the highest concentration (1000 ng/mL), The ρτ elongation by the addition of 150 and 300 ng/mL of the compound (1) was shortened to 17.3 ± 0.2 sec and 24.2 ± 0.3 sec, respectively. The knots of Examples 1 to 3 were obtained. It appears that the shortening effect of the ΡΤ-prolonged ΡΤ produced by the addition of the compound (i) is the gene recombination active factor VII (rFVIIa), and the dried human blood coagulation factor antibody bypass active complex in the order of low to low. [Feiba (registered trademark)], and dried human blood coagulation factor IX complex [PPSB (registered trademark)_HT]. -19- 201219404 (Comparative Example 1) A compound (1), a blood coagulation VIIIB sub-agent [Kogenate (registered trademark) Fs] was added to human plasma, and a blood clotting time automatic rig was used (Amelung KC-10A micro, Thrinity) Biotech Pic) was assayed for activated partial thrombin time (APTT). The plasma concentration in the clinical dose of the blood coagulation factor VIII preparation is estimated to be about 0.44 U/mL [additional file of Kogenate (registered trademark) FS], and in this investigation, a concentration of 0.001 to 6.7 U/mL was used. As a result, among human plasma, APTT was not affected when the VIII factor preparation was coagulated with 0.001 to 1 U/mL of the solution alone, and the compound (1) (150 to 450 ng/mL) was used to extend the APTT in a concentration-dependent manner. . The lU/mL blood coagulation factor VIII preparation shortened the APTT by 300 ng/niL of compound (η with a 1.9-fold extension to 1.6 times. The VIII factor preparation 6.7 was coagulated with a high concentration of blood above 1 U/mL alone. When U/mL, APTT is shortened to 〇.6 times of the control value, and the 2-fold extension by compound (1) is shortened to the control value. (Comparative Example 2) Compounds are added to human plasma (1) The blood coagulation factor IX preparation [Christmassin (registered trademark)-M] was measured for the activated partial thrombin time (APTT) using an automatic blood coagulation time measuring machine (Amelung KC-10A micro, Thrtrlity Biotech Pic). The plasma concentration at the time of administration of 50 U/kg (near clinical use) of Christmassin (registered trademark)-M was estimated to be about 0.47 U/mL, [Christmassin (registered trademark)-Μ attached file], in this discussion, Using the concentration of o.ooi~6.7U/mL" -20- 201219404 As a result, in the human plasma, when the blood is coagulated with the IX factor preparation of lU/rnL alone, it will make the ΑΡΤ·short control value 〇8 Compound (1) will be concentrated at 150~450ng/mL ΑΡΤΤ ΑΡΤΤ 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l l At the most concentration (6.7 U/mL) of blood coagulation factor 制剂 factor preparation, it will shorten to 8 times the control value, and the 2-fold extension by compound (〗) will be shortened to 14 times. (Comparative Example 3) - A compound suspended in 0.5% methylcellulose (ooomg/kg) was orally administered to a rat who was fasted for one night. After 5 minutes of oral administration, the rats were administered with pentobarbital. Anesthetize with thi〇pentai sodium (1〇〇mg/kg, ip), then add fresh plasma to the rats after 30 minutes of oral administration (30mL/kg/2h: 3 times the clinical dose) Continued administration to the jugular vein. The control group continued to administer physiological saline (3〇mL/kg/2h) to the jugular vein. After 2 hours of continuous intravenous infusion of fresh plasma, the lemon was taken from the jugular vein. Acid blood. Plasma is separated from the blood taken, using a blood coagulation time automatic measuring machine (Amelung KC-10A micro, Th Rinity Biotech Pic.) Measured PT. The result 'Compound (1) will prolong ΡΤ to 2.14 times. Fresh plasma (FF) will be produced by compound (1) at a high dose of 3 times the clinical dose. The ΡΤ extension was shortened from 2.14 times to 1.66 times. (Comparative Example 4) Compound (1) (1 〇 mg/kg) suspended in 0.5% methylcellulose was administered to a rat who was fasted for one night, and after 30 minutes, orally Give vitamin Κ 2 (0.1 and 1 mg / kg) or steamed water. After 1 hour and 45 minutes of administration of vitamin Κ2, the rats were anesthetized with sodium pentobarbital (10 mg/kg, i.p.), and after 15 minutes, citrate blood was taken from the lower large vein. The blood was collected from the blood taken out. 'The blood coagulation time automatic measuring machine (Amelung KC-10A micro, Thrinity Biotech Pic.) was used to measure β of pT, and 2.5 hours after oral administration of the compound (1), This will increase ρτ by 1.3 4 times. Vitamin Κ2 does not affect the sputum which is prolonged by the compound (1). (Comparative Example 5) Rats who had fasted for one night were anesthetized with sodium pentobarbital (0. lg/kg, i ρ ). From the start of the anesthesia 15 minutes later to the end of the experiment, the venous vein was continuously administered (1 mg/kg/h) of 5% glucose solution or compound (1), and the bleeding time was measured 2 hours after the start of administration. 8 minutes before the bleeding time was measured, it took 2 minutes to inject physiological saline or tranexamic acid from the thigh vein. The bleeding time was measured by a foot template method or a tail template method. In the sole template method, the rat was placed in a supine posture. On the sole of the right hind limb, a slit of a horizontal straight line having a length of 5 mm was drawn with a cutting blade. The blood is sucked in the filter, paper every 30 seconds, and the time at which the blood becomes no longer infiltrated into the filter paper is measured up to 30 minutes. In the case where hemostasis was not stopped within 30 minutes, the bleeding time was set at 3 minutes. In the tail template method, the rat was placed in a supine position, and a portion of the artery at a position of 4 cm from the front end of the tail was used to draw a cut of 1 mm in depth with a razor. The blood was adsorbed to the filter paper every 15 seconds, and the time during which the blood became no longer infiltrated into the filter paper was measured, up to 3 minutes. This -22-201219404 3 minutes before the start of the measurement of the external bleeding time, a volume ratio of 3.13 ° / tri trisodium citrate dihydrate salt blood sample .5 mL was taken from the jugular vein. Ρτ was measured at 10 min, 4 ° c) after plasma was separated from the blood taken. As a result, the bleeding time of the sole by the administration of the compound (1) 'is significantly prolonged. When 30, 1 and 300 mg/kg of tranexamic acid was administered, it did not show an inhibitory effect on the prolongation of this bleeding time. PT was prolonged to 1.36 times by compound (1), and the same PT elongation was observed in the 30 mg/kg combination of tranexamic acid. Further, by administering the compound (1), the bleeding time in the tail of the rat was remarkably prolonged (the elongation rate was 1.77 times). When 30 and 10 mg/kg of tranexamic acid was administered, it did not show an inhibitory effect on the prolongation of this bleeding time. β ρτ was 1.33 times higher than that of compound 4, and PT was also used in the tranexamic acid combination group. The effect of prolonging the ρτ produced by the administration of the compound (1) was not observed. Furthermore, if the blood cell (Clot) dissolution time measurement system is added to the control group for blood accumulation, then after the blood clot is formed (the absorbance increases), the ghost dissolves rapidly (the absorbance decreases), and the dissolution time is about 4 〇 5 〇 minutes. . In the publication of the publication and U) blood stasis of 0 mg/kg of tranexamic acid, no two receptors were observed until 18 〇 minutes, so the fibrinolytic system (fibrinloytic) whose main effect was confirmed by administration of tranexamic acid System) suppression. From the above results, it seems that 'the use of the rat sole and the tail bleeding type' does not show the acid under the dose of 30, Π) 〇 and 3 延长 for the prolonged bleeding time caused by the compound (1) (1) The reversal effect of the action. σ -23- 201219404 [Industrial Applicability] The reversal of the hydrate of the compound (1), pharmacologically acceptable salt, and 4 it The method of the agent and/or the reversal can reverse the anticoagulant effect of the drug in the event of a bleeding accident, for example, because of a multiplication of 丨; J/L. 【 [Simple description of the figure] Fig. 1 The human blood coagulation factor antibody bypass/tongue complex after drying is shown. The body [Feiba (registered trademark)] and the compound (1) are used alone and in combination with the effect on prothrombin time (PT), and are represented by Fig. 2 is a diagram showing the results of the human blood coagulation π-factor complex [PPSB (registered trademark)·ΗΤ] and the compound (1) after drying, respectively, and in combination with the zymogen time ( Ρτ) the resulting map of influence, and Fig. 3 is a diagram showing the results of Example 2. Fig. 3 is a diagram showing the gene recombinant active type νπ factor (rFvna) and sigma sigma (1) used alone and in combination with prothrombin time (pT) The figure is a diagram, and is a diagram showing the result of the embodiment 3. [Main component symbol description] No 0 - 24-

Claims (1)

201219404 VII. Scope of application: 1. A reversal agent, which is a reversal agent for anticoagulant effect of FXa inhibitors, which is selected from a human blood coagulation factor antibody bypass active complex after drying, and human blood coagulation after drying. In the group consisting of the diterpene factor complex and the gene recombinant active factor VII factor, two or more species are contained. ?h3, n, CH3 2. For the reversal agent of claim 1, wherein the FXa inhibitory drug is Ν1- represented by the following formula (1). - gas acridine_2_yl)_n2-((is,2R,4S)-4-[(dimethylamino)carbonyl]·2-{[(5_methyl_4,5,6,7 _tetrahydrothiazolo[5,4-c]pyridin-2-yl)carbonyl]amino}cyclohexyl)ethanediamine: a pharmacologically acceptable salt, or a hydrate thereof. 3. The reversal agent of claim 1, wherein the FXa inhibitory drug is N1- represented by the formula (la) below. -αpyridin-2-yl)-N2-((is,2R,4S)-4-[(dimethylamino)carbonyl]-2-{[(5-fluorenyl-4,5,6,7 -tetrahydrothiazolo[5,4-C]n-pyridin-2-yl)carbonyl]amino}cyclohexyl)ethanedioxylamine p-toluenesulfonate·1 hydrate: S°3h (la) -25 201219404 4_A kind of N1-(5-aeropyridine-2-yl)_n2-((is,2R,4S)-4-[(didecylamino)carbonyl]-2 -{[(5-Methyl-4,5,6,7-tetrahydrothiazolo[5,4-c]pyridin-2-yl)carbonyl]amino}cyclohexyl)ethanediamine, pharmacologically An acceptable salt or a method for reversing the FXa inhibition of the hydrates, wherein the liquid coagulating factor antibody is coagulated by the IX factor complex, and one or two of the characteristics are: using a human blood circuit selected from the group after drying The active complex, the human blood body after drying, and the gene recombinant active factor VII are added. -26-