HK1155455A - Fxa inhibitors with cyclic amidoxime or cyclic amidrazone as p4 subunit, processes for their preparations, and pharmaceutical compositions and derivatives thereof - Google Patents

Description

FXA inhibitors containing cyclic amidoxime or cyclic amidrazone as P4 subunit, preparation method thereof, and pharmaceutical compositions and derivatives thereof

Technical Field

The present invention relates to novel oxazolidinone derivatives having a cyclic amidoxime or cyclic amidrazone group represented by formula I, pharmaceutically acceptable salts thereof, a preparation method thereof, and pharmaceutical compositions containing the same.

[ formula I ]

Wherein the content of the first and second substances,

ring a is a residue selected from the group consisting of the following structures;

the antithrombotic and anticoagulant effects of the novel oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention are due to the inhibition of an active coagulation protease called factor Xa or other active serine proteases such as factor VIIa, factor IXa or thrombin.

Background

Coagulation factors are distributed in plasma, and various types of factors (coagulation factor 1 to coagulation factor 13) act in a cascade manner to cause blood coagulation. The mechanism in which each coagulation factor participates in blood coagulation is shown in fig. 1.

As shown in fig. 1, blood coagulation is accomplished by a series of very subtle and complex reactions. Usually the unactivated precursor is activated by a specific active coagulation factor (denoted by "a" attached to the end of the coagulation factor). Then, the subsequent coagulation factors are activated. Most of those activated coagulation factors are enzymes of the serine protease family. They adhere to the surface of activated platelets at the wound site and progressively activate coagulation factors, ultimately producing a fibrin clot, resulting in hemostasis.

Thrombin is a multifunctional coagulation factor involved in the end stage of the coagulation cascade. The precursor prothrombin of thrombin is activated via the prothrombinase complex consisting of factor Va, factor Xa, Ca + + and Phospholipids (PL) to produce thrombin, which converts fibrinogen to fibrin. The produced fibrin covers the aggregated platelets to cause blood coagulation. Finally, the fibrin is cross-linked by factor XIIIa to produce a stable fibrin clot.

To generate the prothrombinase (prothrombinase) complex, factor X must be activated to factor Xa, which is mediated primarily by the Xase complex. Factor VIIIa, factor IXa, Ca + + and Phospholipids (PL) produced via the intrinsic pathway or factor VIIa, Tissue Factor (TF) and Ca + + produced via the extrinsic pathway act in the form of a Xase complex.

Thrombin also activates factor V and factor VIII. When thrombin is excessively produced, the blood vessel itself may be occluded. In order to avoid clogging, thrombin initiates blood coagulation inhibition. That is, thrombin binds to thrombomodulin to activate protein C. Activated protein c (apc) complexes with protein S, inactivating factor Va and factor VIIIa.

In fact, factor Xa is itself a serine protease and is involved in the complex blood clotting process. Factor Xa, an essential component of the prothrombinase (prothrombinase) complex, functions as a catalyst for converting prothrombin to thrombin. Thrombin converts fibrinogen into fibrin monomers, which are produced in connection with the production and stabilization of thrombi. Thus, excessive or inappropriate thrombin generation may lead to thromboembolism. Therefore, inhibition of thrombin itself or thrombin generation can reduce fibrin formation associated with thrombosis, preventing thromboembolism.

In short, inhibition of factor Xa results in inhibition of thrombin generation, whereby thromboembolism can be prevented or alleviated. The compounds represented by formula I and pharmaceutically acceptable salts thereof of the present invention inhibit factor Xa, which ultimately prevents thromboembolic disorders (MI, stroke, PE, etc.) according to the above logic.

Of the compounds known as factor Xa inhibitors, antistasin (ats) and Tick Anticoagulant Peptide (TAP) are representative protein inhibitors. ATS (consisting of 119 amino acids) is a natural peptide isolated from leeches with a Ki value for factor Xa of 0.05 nM. TAP is also a peptide consisting of 60 amino acids isolated from ticks and has a Ki value for factor Xa of 0.5 nM. However, these inhibitors have limited use in the clinic; only heparin or its sulfated polysaccharide analogs are used clinically with some limitations.

Low molecular weight compounds have been developed as blood coagulation inhibitors, in particular factor Xa inhibitors, which are described in WO 9529189. Meanwhile, WO9933800 describes factor Xa inhibitors having an indole moiety. In addition, various factor Xa inhibitors have been found and have been in the process of development, for example, heterocyclic compounds containing a nitrogen atom (WO2004058743), imidazole derivatives (WO2004050636), pyrazole derivatives (WO2004056815), indole-2-carboxamide derivatives (WO2003044014), oxybenzamide (oxybenzamide) derivatives (WO2002051831), guanidine/amidine derivatives (WO2002046159), amino-bicyclic pyrazinone/pyridone derivatives (WO2004002405), and the like.

To be clinically effective, these molecules should have, as FXa inhibitors, a high antithrombotic effect, high stability, a suitable selectivity for other related serine proteases (thrombin, trypsin, cathepsin G, etc.), low toxicity and satisfactory bioavailability in both plasma and liver.

The latest compound with oxazolidinone, similar to the compounds of the present invention, is rivaroxaban (formula a), which is under clinical phase III evaluation. Some oxazolidinone derivatives represented by formula 2 are described in WO 01/47917. However, some of these compounds are reported to have limited solubility; a specific example of this problem is rivaroxaban. The solubility of rivaroxaban was only 8 mg/L. Poor solubility can create many practical limitations (including variability and slow dissolution). These problems can be overcome by introducing a highly soluble moiety.

[ formula A ]

[ formula B ]

X＝CH₂，O，S，N-R

Y＝-NCOR′

WO 2004/83174 describes the use of pyrazole derivatives including Apixaban. Some of these inhibitors are cyclic amidine and sulfonylamidine derivatives represented by formula C.

However, there is no previous example similar to the present invention describing the specific incorporation of cyclic amidoximes or cyclic amidrazones in the oxazolidinone backbone as factor Xa inhibitors. In fact, little is known about the substantiation of cyclic amidoximes or cyclic amidrazones in drug design.

[ formula C ]

The main trend in recent research on FXa and thrombin inhibitors is the implementation of the amidine function. The amidine function (the so-called P1 group) is designed to bind Asp at the bottom of the S1 pocket¹⁸⁹. Both FXa and thrombin recognize the arginine residue in the natural substrate as the P1 site. The amidine group (including guanidine derivatives) that replaces guanidine in arginine is highly hydrophilic. Thus, inhibitors containing amidine functional groups are generally poorly absorbed and, even if they are absorbed, they are cleared too quickly due to the high polarity inherent in amidines (Drugs of the Future, 1999, 24(7), 771).

Amidines themselves have a strong basicity (PKa: about 12.5). Amidine inhibitors generally exhibit poor absorption under physiological conditions due to formal positive charge. Therefore, there is a need to change to a less basic alternative. Representative examples of these are pyridine derivatives, amidrazones, cyclic amines, alkylamine derivatives, aminobenzisoxazoles, and the like (US 6958356). There are also fundamentally different approaches to overcome these problems, including amidoximes. Amidoximes, which are prodrugs based on the fact that weak N-O bonds in vivo are readily reduced to amidines, are readily synthesized by introducing hydroxyl groups into the amidine structure. The PKa (8-9) of the process using amidoximes is significantly lower than that of amidines. Ximelagatran is another example of a prodrug of this type. This trend was seen not only in the study of FXa inhibitors but also in the study of thrombin inhibitors. However, most of these attempts did not yield as good as expected. As a third attempt, a neutral P1 group was introduced. Unlike other types of drugs, FXa and thrombin inhibitors tend to exhibit good effectiveness at high concentrations in the blood. In addition, the concentration of free drug that is not bound to serum proteins in the blood is very important. In the case of neutral P1 group inhibitors, the tendency to bind to proteins is high, resulting in less than expected effectiveness.

To overcome these problems, the present inventors introduced a relatively polar group at a position other than the P1 site and fixed a neutral group at the P1 site. Similar logic, including some other important factors that improve efficacy as follows: 1) the water solubility is greatly improved; 2) low binding to plasma proteins; when the free concentration of the drug is high, the potency in the PT assay is also increased even though the FXa affinity is slightly reduced.

In the present invention, the selected site for introducing the polar group is the P4 subsite (sub-site) of the inhibitor, and the logic of this selection is as follows. The S4 site of FXa has a U-shaped binding site surrounded on three sides by Tyr99, Phe174 and Trp 215. This binding site is composed only of aromatic amino acid side chains, unlike the case of thrombin surrounded by Leu99, lle174, and Trp 215. This difference is exploited in drug design.

The S4 pocket of FXa interacts efficiently with cationic residues, which is commonly referred to as "pi-cation interaction. In fact, some inhibitors with a positive charge at the P4 site were designed and synthesized. In the present invention, as described above, in order to improve water solubility and increase drug effectiveness by reducing binding to proteins, cyclic amidoxime or amidrazone is introduced at position P4. The ring form was chosen because the inventors believe that it is possible to improve absorption by reducing the number of NH bonds, which generally shows a negative effect on absorption. According to recent studies, it is more advantageous to have less Hydrogen Bond Donors (HBD) for drugs relative to Hydrogen Bond Acceptors (HBA). According to the Lipinski rule, up to 10 HBAs may be acceptable, but the HBD is limited to only up to 5 (adv. drug Delivery rev., 2001, 46, 3-26), especially in the case of new drugs, the average number of HBDs is about 2, indicating that the HBD is more severely limited (j.med. chem.2004, 47, 6338-48). The amidoxime or amidrazone group itself has basicity, which makes it easier to isolate-purify-store in the form of a salt, and thus, it is expected that water solubility will increase.

Briefly, we introduced an amidoxime or amidrazone at position P4. To reduce the HBD number, we used the ring form of the functional group to prepare more drug-like (drug-like) inhibitors.

The compounds of formula I according to the invention have proven to have the above-mentioned advantages. Water solubility and protein binding levels are shown, as well as 2x PT values and Ki.

Disclosure of Invention

Technical problem

The present inventors have synthesized novel oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group with useful properties, which can be used to prepare pharmaceutical formulations. In particular, oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group exhibit FXa inhibitory effects, and thus they are useful for the treatment or prevention of thrombosis, myocardial infarction, arteriosclerosis, inflammation, stroke, angina pectoris, restenosis after angioplasty, and thromboembolism such as intermittent claudication. Furthermore, the oxazolidinone derivatives containing said cyclic amidoxime or cyclic amidrazone of the invention can be used as inhibitors of factor VIIa, factor IXa and thrombin, which are procoagulant factors in the blood coagulation cascade.

It is an object of the present invention to provide novel oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group or pharmaceutically acceptable salts thereof which exhibit factor Xa inhibitory properties.

It is another object of the present invention to provide a pharmaceutical composition for anticoagulation, which comprises, as an active ingredient, an oxazolidinone derivative containing a cyclic amidoxime or cyclic amidrazone group, or a pharmaceutically acceptable salt thereof.

It is another object of the present invention to provide a pharmaceutical composition comprising an oxazolidinone derivative having a cyclic amidoxime or cyclic amidrazone group or a pharmaceutically acceptable salt thereof as an active ingredient for the treatment or prevention of thrombosis, myocardial infarction, arteriosclerosis, inflammation, stroke, angina pectoris, restenosis, intermittent claudication, venous thrombosis, pulmonary embolism, arterial thrombosis, myocardial ischemia or thromboembolism.

It is another object of the present invention to provide a pharmaceutical composition for the therapeutic or prophylactic treatment of coronary artery diseases, cerebral artery diseases and peripheral artery diseases, which is characterized by co-treating (co-treat) an oxazolidinone derivative having a cyclic amidoxime or cyclic amidrazone group or a pharmaceutically acceptable salt thereof with a thrombolytic agent.

It is another object of the present invention to provide the use of oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group or pharmaceutically acceptable salts thereof as anticoagulants for the extracorporeal preservation of blood, plasma or blood products.

Detailed Description

The present invention relates to novel oxazolidinone derivatives having a cyclic amidoxime or cyclic amidrazone group represented by formula I, or pharmaceutically acceptable salts thereof, a preparation method thereof, and a pharmaceutical composition comprising the same.

[ formula I ]

Wherein ring a is a residue selected from the group consisting of the following structures;

R₁to R₁₂Independently H, (C)₁-C₇) Alkyl, (C)₃-C₇) Cycloalkyl group, (C)₆-C₁₂) Aryl or (C)₄-C₁₂) Heteroaryl containing 1-4 heteroatoms selected from O, S and N, R₃And R₄Through the reaction of (C)₃-C₅) The alkylene group is linked to form a ring, the carbon atom of the alkylene group may be substituted with a carbonyl group, and R₁To R₁₂The alkyl, cycloalkyl, aryl or heteroaryl of (A) may be selected from (C)₁-C₇) Alkyl, halo (C)₁-C₇) Alkyl, (C)₁-C₇) One of alkoxy and halogen;

R₁₃is H, (C)₁-C₇) Alkyl, (C)₃-C₇) Cycloalkyl, formyl, (C)₁-C₇) Alkylcarbonyl group, (C)₁-C₇) Alkoxycarbonyl or (C)₆-C₁₂) And (4) an aryl group.

In the present invention, "aryl" is an organic group obtained by removing 1H from an aromatic hydrocarbon, wherein each ring of a monocyclic or fused ring system contains 6 to 12 ring atoms, preferably 6 to 10 ring atoms. Specifically, it includes phenyl, naphthyl, biphenyl and indenyl groups, but is not always limited thereto.

"heteroaryl" in the present invention means an aryl group containing 1 to 4 heteroatoms selected from N, O and S as aromatic ring structural atoms and the remaining aromatic ring structural atoms being C, and examples thereof are 5-6-membered monocyclic heteroaryl, and polycyclic heteroaryl fused with one or more partially saturated benzene rings.

The oxazolidinone derivatives containing cyclic amidoxime or cyclic amidrazone group of the present invention are selected from the following formulas II to XI.

[ formula II ]

[ formula III ]

[ formula IV ]

[ formula V ]

[ formula VI ]

[ formula VII ]

[ formula VIII ]

[ formula IX ]

[ formula X ]

[ formula XI ]

Wherein R is₁To R₁₂Independently H, (C)₁-C₇) Alkyl or (C)₃-C₇) A cycloalkyl group; r₁₃Is H, (C)₁-C₇) Alkyl, (C)₃-C₇) Cycloalkyl, formyl or (C)₁-C₇) An alkylcarbonyl group; and m is an integer of 1 to 3.

As examples of the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group according to the present invention, R is represented by the following formulas II to XI₁To R₁₂Independently H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; r₁₃Is H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, formyl or acetyl; and m is an integer of 1.

The oxazolidinone derivative having a cyclic amidoxime or cyclic amidrazone group according to the present invention may be exemplified by the following compounds, but is not always limited thereto.

In the present invention, the preparation method of oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I is exemplified by reaction schemes 1 to 5. However, the following method does not limit the preparation method of the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention, and it is well understood by those skilled in the art that modifications of the following method are allowed and also included in the scope of the present invention. Unless otherwise indicated, substituents in the reaction schemes are as defined in formula I.

As shown in reaction schemes 1 to 5, the preparation method of the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I is changed according to the structure of the cyclic compound a in formula I. First, cyclic amidoxime compounds, such as compounds B1 and B2 of reaction scheme 1 and compound E of reaction scheme 4, are synthesized. Cyclic amidrazone compounds, such as compounds C1 and C2 of reaction scheme 1, compound D1 of reaction scheme 2, and compound F of reaction scheme 5, were also synthesized.

[ reaction scheme 1]

As shown in reaction scheme 1, the synthesis of cyclic amidoxime compounds B1 and B2 and cyclic amidrazone compounds C1 and C2 of formula I was performed by reacting 4-cyanoaniline (1) and 2- (((S) -oxiran-2-yl) methyl) tert-butoxycarbonyl (2) to give compound 3. Oxazolidinone ring compound 4 was then synthesized using 1, 1-carbonyldiimidazole and DMAP, which was treated with HCl to remove the boc protecting group. Condensation with 5-chlorothiophene-2-carboxylic acid, followed by treatment with HCl. Finally, this reactant is reacted with a diamine compound to give the cyclic amidoxime compounds B1 and B2 and cyclic amidrazone compounds C1 and C2 of formula I.

In the synthesis of compound 3, similar to the procedure of reaction scheme 2, the phthalimide-protected amine compound can be prepared by using 2- (((S) -oxiran-2-yl) methyl) isoindoline-1, 3-dione (9) of reaction scheme 2 instead of 2- (((S) -oxiran-2-yl) methyl) tert-butoxycarbonyl (2). In reaction scheme 1, a process using 2- (((S) -oxirane-2-yl) methyl) tert-butoxycarbonyl (2) is described, and in reaction scheme 2, a process using 2- (((S) -oxirane-2-yl) methyl) isoindoline-1, 3-dione (9) is described.

The cyclic amidrazone compounds D1 and D2 of formula I may be divided according to the position of the double bond and synthesized according to reaction scheme 2 and reaction scheme 3.

[ reaction scheme 2]

First, 4-nitroaniline (6) was protected with a boc group, and then hydrogenated with a palladium catalyst. Thereafter, the amino alcohol compound 10 was synthesized using 2- (((S) -oxirane-2-yl) methyl) isoindoline-1, 3-dione (9). Preparation of the oxazolidone ring using carbonyl diimidazole affords compound 11. The phthalimide protecting group is removed by hydrazine and then condensed with 5-chlorothiophene-2-carboxylic acid to give compound 13. Compound 13 throughHCl treatment to remove the boc protecting group, followed by reaction with a boc protected aminal affords compound 14. With NaNO₂Introduction of a nitroso group followed by reduction with Zn affords the hydrazine compound 15. Reaction of compound 15 with orthoformate affords the cyclic amidrazone compound D1.

Another cyclic amidrazone compound D2 was synthesized according to reaction scheme 3.

[ reaction scheme 3]

Synthesis of compounds 14 shown in scheme 2 wherein R₇Compound 14a, which is H, to which amino groups (15a) are introduced, followed by stepwise introduction of alkyl groups (16). Cyclization with orthoformate affords cyclic amidrazone compound D2.

Cyclic amidoxime compounds E of formula I were synthesized by reaction scheme 4.

[ reaction scheme 4]

Compound 17, prepared from 4-fluoronitrobenzene, is reacted with methanesulfonyl chloride to give compound 18, which is then reacted with hydroxyphthalimide to give compound 19. Removal of the phthalimide protecting group by hydrazine, followed by protection with a boc group, affords compound 21. The nitro group of compound 21 is reduced by Zn to give compound 22, which is then reacted with 2- (((S) -oxiran-2-yl) methyl) isoindoline-1, 3-dione (9) according to the method shown in reaction scheme 2 to give compound 26. Compound 26 is treated with HCl and then reacted with orthoester to give compound E.

The cyclic amidrazone compounds F of formula I were synthesized by reaction scheme 5.

[ reaction scheme 5]

Protection of Compound 17 from 4-fluoronitrobenzene with a tbs (t-butyldimethylsilyl) group followed by protection of the amine moiety with boc affords Compound 27. The amine was prepared using a palladium catalyst and reacted with Cbz-Cl to provide compound 28. Compound 28 was reacted with glycidyl butyrate to give compound 29. Replacement of the alcohol group with an amine gives compound 30. This compound was reacted with chlorothiophene carboxylic acid to give compound 31, which was then reacted with methanesulfonyl chloride. This reaction was treated with hydrazine to afford compound 33. Compound 33 is reacted with an orthoester to give compound F.

The oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention can be used as pharmaceutically active ingredients, particularly for the prevention and treatment of thrombosis, myocardial infarction, arteriosclerosis, inflammation, stroke, angina pectoris, restenosis after angioplasty, intermittent claudication, venous thrombosis, pulmonary embolism, arterial thrombosis, myocardial ischemia, unstable angina due to thrombosis, and thromboembolism such as crisis in a medicine.

The oxazolidinone derivative containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention or a pharmaceutically acceptable salt thereof may be used for preventing or treating atherosclerotic diseases including coronary artery diseases, cerebral artery diseases, or peripheral artery diseases. For the treatment of myocardial infarction, the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group may be used in combination with a thrombolytic agent (e.g., alteplase, tenecteplase, etc.). The compounds may also be used to prevent reocclusion following thrombolysis, Percutaneous Transluminal Coronary Angioplasty (PTCA), and coronary artery bypass surgery.

The oxazolidinone derivative containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention or a pharmaceutically acceptable salt thereof can be used to prevent postoperative thrombosis from being re-formed. The compounds may also be used as anticoagulants in connection with artificial organs or hemodialysis. The compounds may be used for cleaning catheters and medical accessories used in the body. In addition, the compounds may also be used as anticoagulant compositions for ex vivo storage of blood, plasma and other types of blood products. The compounds of the present invention are also effective in treating diseases associated with blood coagulation or diseases that cause secondary damage such as cancer (including metastatic cancer), inflammation including arthritis and diabetes.

The oxazolidinone derivative containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention may be used in the form of a pharmaceutically acceptable salt. For pharmaceutically acceptable salts, acid addition salts prepared using pharmaceutically acceptable free acids are preferred. Either inorganic or organic free acid may be used as long as it is a pharmaceutically acceptable free acid. Examples of inorganic free acids include: hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid. Examples of organic free acids that can be obtained are: citric acid, acetic acid, lactic acid, maleic acid, fumaric acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, trifluoroacetic acid, galacturonic acid, pamoic acid, glutamic acid, and aspartic acid. The oxazolidinone derivative of the present invention may include a hydrate of a salt. In particular, if the salt is hygroscopic, it is preferably used in the form of a crystalline hydrate.

The oxazolidinone derivative containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention may be formulated in the form of a prodrug designed to increase absorption or solubility in vivo, and may be used in the form of a hydrate or solvate. For example, as described below, the attachment is a group that can be easily isolated after absorption in the body, or the compound is prepared in the form of a salt, specifically, in the form of one or more hydrates or solvates. Prodrugs, as well as hydrates or solvates of the salts, are also included within the scope of the present invention.

An effective dose of the oxazolidinone derivative represented by formula I, a hydrate thereof, a solvate thereof, or a pharmaceutically acceptable salt thereof may be determined to produce a therapeutic effect in consideration of the specific compound used, the administration method, the target subject, the target disease, and the like, but 0.1 to 20mg/kg (body weight) per day is a preferred dose of the oxazolidinone derivative compound represented by formula I. The daily dose may be administered once daily (once), or may be divided into several administrations daily in suitably divided forms within the daily effective dose. According to the dosage form, oral administration, parenteral (injection) administration or topical administration may be permitted. The pharmaceutical composition of the present invention may be formulated for oral administration, such as tablets, powders, dry syrups, chewable tablets, granules, capsules, soft capsules, pills, drinks, sublingual agents and the like. The compositions of the present invention formulated as tablets may be administered to a subject by any method or route (which may be the oral route) that has bioavailability that delivers an effective dose of the tablet. And the method or route of administration may be determined according to the characteristics, stage and other conditions of the disease of interest. When the composition of the present invention is formulated into a tablet, it may additionally comprise a pharmaceutically acceptable excipient. The amount and characteristics of the excipients may be determined according to the solubility and chemical properties of the selected tablet, the route of administration and standard pharmaceutical practice.

Drawings

The above and other objects, features and advantages of the present invention will become apparent from the following description of preferred embodiments thereof, when taken in conjunction with the accompanying drawings, in which:

fig. 1 is a diagram illustrating the mechanism of blood coagulation.

Examples

Practical and presently preferred embodiments of the present invention are shown in the following examples.

However, it will be appreciated that those skilled in the art, upon consideration of this disclosure, may make modifications and improvements within the spirit and scope of the present invention.

Preparation example 1: preparation of Compound 5

<1-1>Preparation of Compound 3

4-aminobenzonitrile (1) (5g, 42.30mmol) and 2- (((S) -Oxiran-2-yl) methyl) tert-butoxycarbonyl (2) (8.79g, 50.78mmol) were added to 2-isopropanol (20mL), followed by stirring and refluxing for 12 hours. The reaction was concentrated under reduced pressure and then column treated to give the title compound 3 as a white solid (7.30g, 25.1mmol, 59%).

¹H NMR (400MHz, chloroform-d)₁)δ＝7.41(d，J＝8.4Hz，4H)，6.59(d，J＝8.4Hz，1H)，4.95(br s，1H)，4.80(br s，1H)，3.97-3.93(m，1H)，3.31-3.15(m，5H)，1.46(s，9H)

<1-2>Preparation of Compound 4

Compound 3(7.30g, 25.05mmol) obtained above, 1-carbonyldiimidazole (4.87g, 30.06mmol) and dimethylaminopyridine (1.53g, 12.52mmol) were gradually added to tetrahydrofuran (70mL), followed by stirring and refluxing for 12 hours. The reaction was concentrated under reduced pressure and then dissolved in ethyl acetate (300 mL). After washing stepwise with 1N-HCl solution (50mL) and sodium bicarbonate solution (50mL), the reaction was dried over sodium sulfate and then concentrated under reduced pressure. The reaction was washed with diethyl ether (100mL) to afford compound 4 as a white solid (6.60g, 20.8mmol, 83%).

¹H NMR (400MHz, chloroform-d)₁)δ＝7.67(s，4H)，4.95(br s，1H)，4.82-4.79(m，1H)，4.07(dd，J＝8.8，8.8Hz，1H)，3.94(dd，J＝8.8，6.8Hz，1H)，3.56-3.54(m，2H)，1.38(s，9H)

<1-3>Preparation of Compound 5

Compound 4(6g, 18.90mmol) obtained above was added to ethyl acetate (10mL), which was added to 4N-HCl dissolved in 1, 4-dioxane (60mL), followed by stirring at room temperature for 1 hour. The resulting solid was filtered under reduced pressure and then washed stepwise with ethyl acetate (20mL) and diethyl ether (30 mL). This gave the hydrochloride salt of the amine compound without boc as a white solid (4.60g, 18.1mmol, 95.9%).

¹H NMR (400MHz, chloroform-d)₁)δ＝8.36(br s，3H)，7.90(d，J＝8.8Hz，2H)，7.73(d，J＝8.8Hz，2H)，5.03-4.96(m，H)，4.25(dd，J＝9.2，9.2Hz，1H)，3.92(dd，J＝9.2，6.4Hz，1H)，3.28-3.25(m，2H)

The amine compound (4.60g, 18.13mmol), HOBt (2.75g, 19.94mmol), EDC (4.17g, 21.75mmol), 5-chlorothiophene-2-carboxylic acid (3.20g, 19.04mmol) and triethylamine (5.70mL, 39.88mmol) were added stepwise to N, N-dimethylformamide (50mL) and stirred at room temperature for 12 hours. This reaction was slowly added to distilled water (800mL), and the resulting solid was filtered under reduced pressure. The reaction was washed with diethyl ether (100mL) to afford compound 5 as a white solid (5.70g, 15.8mmol, 87%).

¹H NMR(400MHz，DMSO-d₆)δ＝8.93(t，J＝5.2Hz，1H)，7.81(d，J＝9.2Hz，2H)，7.69(d，J＝9.2Hz，2H)，7.63(d，J＝4.0Hz，1H)，7.15(d，J＝4.0Hz，1H)，4.86-4.81(m，1H)，4.17(dd，J＝9.2，9.2Hz，1H)，3.83(dd，J＝9.2，5.2Hz，1H)，3.57(dd，J＝5.2，5.2Hz，2H)

LCMS：362(M+H⁺)，C₁₆H₁₂ClN₃O₃S

Preparation example 2: preparation of Compound 13

<2-1>Preparation of Compound 7

4-nitroaniline (20g, 145mmol) was dissolved in acetonitrile (200mL), and di-tert-butyldicarbonate (63.2g, 290mmol) and 4-dimethylaminopyridine (3.54g, 29mmol) were added thereto, followed by stirring and refluxing for 16 hours. The reaction solution was cooled at room temperature and then concentrated under reduced pressure to give a brown solid compound containing two boc groups (49g, 145mmol, 100%).

¹H NMR(600MHz，CDCl₃)δ＝8.25(d，J＝9Hz，2H)，7.36(d，J＝9Hz，2H)，1.45(s，18H)

The resulting compound (49g, 145mmol) was dissolved in methanol (200mL), to which was added potassium carbonate (60g, 434mmol), followed by stirring and refluxing for 16 hours. The reaction solution was cooled at room temperature and then concentrated under reduced pressure. The reaction solution was subjected to column chromatography (n-hexane/ethyl acetate, 6/1) to give the title compound 7 as a pale yellow solid (17.6g, 73.9mmol, 51%).

¹H NMR(600MHz，CDCl₃)δ＝8.18(d，J＝9Hz，2H)，7.53(d，J＝9Hz，2H)，6.93(br s，1H)，1.54(s，9H)

<2-2>Preparation of Compound 8

Compound 7(17.6g, 73.9mmol) was dissolved in ethyl acetate (200mL), to which was added palladium on carbon (10 wt%, 3.9g), followed by stirring in a hydrogen balloon. After 16 h, the reaction solution was filtered through magnesium fluoride then concentrated under reduced pressure to give the title compound 8 as a pale pink solid (15.4g, 73.9mmol, 100%).

¹H NMR(600MHz，CDCl₃)δ＝7.12(br s，2H)，6.62(d，J＝9Hz，2H)，6.31(br s，1H)，3.53(br s，2H)，1.50(s，9H)

<2-3>Preparation of Compound 10

Compound 8(13.5g, 65.1mmol) was dissolved in 2-propanol (170mL), and (S) -glycidylphthalimide (9) (14.6g, 71.9mmol) was added thereto, followed by continuing the reaction for 12 hours. Then (S) -glycidylphthalimide (9) (2.65g, 13.0mmol) was additionally added, followed by stirring and refluxing for 4 hours. The reaction solution was cooled at room temperature and then concentrated under reduced pressure. Recrystallization from n-hexane (500mL) gave the title compound 10 as a yellow solid (26.8g, 65.1mmol, 100%).

¹H NMR(600MHz，CDCl₃)δ＝7.89-7.84(m，2H)，7.78-7.73(m，2H)，7.15(br，2H)，6.63(d，J＝8Hz，2H)，6.26(br，1H)，4.18-4.12(m，1H)，4.05(br，1H)，3.94-3.86(m，2H)，3.25(dd，J＝13，4.5Hz，1H)，3.15(dd，J＝13，6.6Hz，1H)，2.84(d，J＝4.8Hz，1H)，1.50(s，9H)

<2-4>Preparation of Compound 11

Compound 10(26.8g, 65.1mmol) was dissolved in tetrahydrofuran (200mL), to which 1, 1-carbonyldiimidazole (15.9g, 98.1mmol) and 4-dimethylaminopyridine (1.59g, 13.0mmol) were added, followed by stirring and refluxing for 16 hours. The reaction solution was cooled and then concentrated under reduced pressure. Saturated ammonium chloride solution (200mL) was added, followed by extraction with ethyl acetate (250 mL. times.2). The collected organic layers were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then treated by column chromatography (n-hexane/ethyl acetate/dichloromethane, 1/1/1) to give the title compound 11 as a pale yellow solid (20.0g, 45.7mmol, 70%).

¹H NMR(600MHz，CDCl₃)δ＝7.91-7.87(m，2H)，7.79-7.74(m，2H)，7.43(d，J＝9Hz，2H)，7.36(d，J＝9Hz，2H)，6.48(br s，1H)，5.00-4.95(m，1H)，4.15(dd，J＝14，7Hz，1H)，4.11(t，J＝9Hz，1H)，3.97(dd，J＝14，6Hz，1H)，3.89(dd，J＝9，6Hz，1H)，1.52(s，9H)

<2-5>Preparation of Compound 12

Compound 11(15.3g, 35.0mmol) was dissolved in ethanol (200mL), to which was added hydrazine hydrate (3.40mL, 70.0mmol), followed by stirring and refluxing for 3 hours. The reaction solution was cooled at room temperature. The resulting white solid was filtered off, and the filtrate was concentrated under reduced pressure. Methylene chloride (100mL) was added thereto, and the resultant solid was removed by filtration. The filtrate was concentrated under reduced pressure. This procedure was repeated two more times, then the reaction was dried to give the title compound 12 as a white solid (10.0g, 32.5mmol, 93%).

¹H NMR(600MHz，CDCl₃)δ＝7.45(d，J＝9Hz，2H)，7.37(d，J＝9Hz，2H)，6.65(br s，1H)，4.67-4.63(m，1H)，4.03(t，J＝9Hz，1H)，3.82(dd，J＝9，7Hz，1H)，3.09(dd，J＝14，4Hz，1H)，2.98(dd，J＝14，6Hz，1H)，1.52(s，9H)

<2-6>Preparation of Compound 13

Compound 12(3.63g, 11.8mmol) was dissolved in chloroform (50mL), to which was added 5-chlorothienylcarboxylic acid (2.30g, 14.1mmol) and 4-dimethylaminopyridine (0.30g, 13.0 mmol). The temperature is reduced to 0 ℃. N, N' -diisopropylcarbodiimide (2.20mL, 14.1mmol) was added thereto, followed by stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and then recrystallized from an n-hexane/diethyl ether solution (1/1, 200mL) to give the title compound 13 as a white solid (5.0g, 11.1mmol, 94%).

¹H NMR(400MHz，DMSO-d₆)δ＝9.30(s，1H)，8.95(t，J＝6Hz，1H)，7.67(d，J＝4Hz，1H)，7.45-7.36(m，4H)，7.17(d，J＝4Hz，1H)，4.82-4.74(m，1H)，4.11(t，J＝9Hz，1H)，3.77(dd，J＝9，6Hz，1H)，3.57(t，J＝5.6Hz，2H)，1.45(s，9H)

Preparation example 3: preparation of Compound 16a

<3-1>Preparation of Compound 14a

Compound 13(16.5g, 36.5mmol) was dissolved in dichloromethane (150mL), to which HCl (150mL, 4M 1, 4-dioxane solution) was added, followed by stirring at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, followed by drying to obtain a white solid compound (14.1g, 36.3mmol, 99%).

¹H NMR(600MHz，DMSO-d₆)δ＝9.07(t，J＝6Hz，1H)，7.73(d，J＝3.6Hz，1H)，7.63(d，J＝9Hz，2H)，7.37(d，J＝9Hz，2H)，7.20(d，J＝3.6Hz，1H)，4.88-4.83(m，1H)，4.18(t，J＝9Hz，1H)，3.87(dd，J＝9，6Hz，1H)，3.61(t，J＝5.4Hz，2H)

Methanol (40mL) and N, N-dimethylformamide (15mL) were added to the compound (3.0g, 7.73mmol) obtained above. N-Boc-2-aminoacetaldehyde (1.48g, 9.30mmol) and sodium cyanoborohydride (486mg, 7.73mmol) were added thereto, followed by stirring at room temperature for 16 hours. Saturated ammonium chloride solution (20mL) was added thereto, and then the solvent was concentrated under reduced pressure. Saturated ammonium chloride solution (50mL) was added, followed by extraction with ethyl acetate (250 mL. times.2). The collected organic layers were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then treated by column chromatography (n-hexane/ethyl acetate, 1/2 → 1/4) to give the title compound 14a as a white solid (2.76g, 5.58mmol, 72%).

¹H NMR(400MHz，CDCl₃)δ＝7.30(d，J＝4Hz，1H)，7.18(d，J＝9Hz，2H)，7.00(t，J＝6Hz，1H)，6.80(d，J＝4Hz，1H)，6.53(d，J＝9Hz，2H)，4.84-4.74(m，2H)，4.04(br，1H)，3.98(t，J＝9Hz，1H)，3.81(ddd，J＝14.4，6，3Hz，1H)，3.74(dd，J＝9，6Hz，1H)，3.66(dt，J＝14.8，9Hz，1H)，3.36-3.26(m，2H)，3.18(t，J＝6Hz，2H)，1.41(s，9H)

<3-2>Preparation of Compound 15a

Compound 14a (1.0g, 2.0mmol) was dissolved in acetic acid (10mL) and sodium nitrate (NaNO) dissolved in distilled water (2mL) was slowly added thereto at 0 deg.C₂) (170mg, 2.46mmol) and then stirred for 30 minutes. Then, zinc-amalgam (650mg of zinc washed with 0.5% mercury (II) acetate solution, then distilled water, used directly) was added thereto, followed by stirring at 0 ℃ for 5 hours. Saturated sodium carbonate solution (50mL) was added slowly, followed by extraction with ethyl acetate (250mL x 2). The collected organic layers were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then treated by column chromatography (n-hexane/ethyl acetate, 1/2 → 1/4) to give the title compound 15a as a pale yellow solid (450mg, 0.88mmol, 45%).

¹H NMR(600MHz，CDCl₃)δ＝7.34(d，J＝3.6Hz，1H)，7.32(d，J＝8.4Hz，2H)，6.96(d，J＝8.4Hz，2H)，6.90(br，1H)，6.87(d，J＝3.6Hz，1H)，4.94(br s，1H)，4.87-4.81(m，1H)，4.05(t，J＝9Hz，1H)，3.90-3.83(m，1H)，3.80(dd，J＝9，6Hz，1H)，3.75-3.67(m，1H)，3.68(br s，2H)，3.44(br，4H)，1.41(s，9H)

<3-3>Preparation of Compound 16a

Compound 15a (150mg, 0.29mmol) was dissolved in methanol (3mL), to which formalin (0.10mL, 37 wt% aqueous solution) was added, followed by stirring at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, then diluted with distilled water (15mL), followed by extraction with dichloromethane (15 mL. times.2). The collected organic layers were dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure to give 110mg of a pale yellow solid. This solid was dissolved in methanol (3mL) and tetrahydrofuran (1mL) and then cooled to 0 ℃. To this was added sodium borohydride (160mg, 4.22 mmol). The pH was adjusted to 5 with acetic acid and the reaction temperature was slowly raised to 50 ℃. After 10 hours, the reaction solution was concentrated under reduced pressure, diluted with 2N HCl solution (30mL), and extracted with dichloromethane (25 mL. times.2). The collected organic layers were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then treated by column chromatography (n-hexane/ethyl acetate, 1/1 → 1/3) to give the title compound 16a as a white solid (15mg, 0.029 mmol).

Preparation example 4: preparation of Compound 15b

<4-1>Preparation of Compound 14b

Compound 13 was dissolved in dichloromethane as described in the synthesis of compound 14a in preparative example 3, to which HCl (4M 1, 4-dioxane solution) was added, followed by stirring at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, followed by drying to give a white solid compound (500mg, 1.29 mmol). Methanol (10mL) and N, N-dimethylformamide (2mL) were added thereto, and N-Boc-N-methyl-2-aminoacetaldehyde (268mg, 1.55mmol) and sodium cyanoborohydride (81mg, 1.29mmol) were added thereto, followed by stirring at room temperature for 16 hours. Saturated ammonium chloride solution (3mL) was added thereto, and then the solvent was concentrated under reduced pressure. Saturated ammonium chloride solution (30mL) was added thereto, followed by extraction with ethyl acetate (30mL × 2). The collected organic layers were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then treated by column chromatography (n-hexane/ethyl acetate, 1/2 → 1/4) to give the title compound 14b as a white solid (544mg, 1.07mmol, 83%).

¹H NMR(600MHz，CDCl₃)δ＝7.31(d，J＝4Hz，1H)，7.24(d，J＝9Hz，2H)，6.89(d，J＝4Hz，2H)，6.65(br，1H)，6.59(d，J＝9Hz，1H)，4.85-4.79(m，1H)，4.04(t，J＝9Hz，1H)，3.90(ddd，J＝16.6，7，3Hz，1H)，3.79(dd，J＝9，6Hz，1H)，3.74-3.67(m，1H)，3.54-3.39(m，2H)，3.26(t，J＝6Hz，2H)，2.88(s，3H)，1.46(s，9H)

<4-2>Preparation of Compound 15b

Compound 14b (405mg, 0.80mmol) was dissolved in acetic acid (3mL) and sodium nitrate (NaNO) dissolved in distilled water (0.5mL) was slowly added thereto at 0 deg.C₂) (66mg, 0.96mmol) and then stirred for 30 minutes. Thereafter, zinc (130mg, 1.99mmol) was added thereto, followed by stirring at 0 ℃ for 3 hours. Saturated sodium carbonate solution (30mL) was added slowly, followed by extraction with ethyl acetate (30mL × 2). The collected organic layers were dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and then treated by column chromatography (n-hexane/ethyl acetate, 1/2 → 1/4) to give the title compound 15b as a pale brown solid (81mg, 0.15mmol, 19%).

Preparation example 5: preparation of Compound 26

<5-1>Preparation of Compound 18

4-Fluoronitrobenzene (5.2g, 37mmol) was dissolved in acetonitrile (40mL), 2-aminoethanol (5.2g, 85mmol) was added thereto, followed by stirring overnight and refluxing. The reaction solution was cooled at room temperature, concentrated under reduced pressure, dissolved in ethyl acetate and washed stepwise with water, 1N HCl and brine. The reaction was dried over anhydrous sodium sulfate, filtered, and then distilled under reduced pressure to obtain compound 17. Compound 17(15g, 82.33mmol) and diisopropylethylamine (27mL, 164mmol) were dissolved in dichloromethane (150mL), to which methanesulfonyl chloride (9.5mL) was slowly added at 0 ℃ and then stirred at room temperature for 2 hours. Upon completion of the reaction, dichloromethane (800mL) was added to the reaction solution. The reaction was washed with sodium bicarbonate solution (500mL) and then concentrated under reduced pressure to give compound 18(22g, 82.00mmol, 99%) as a yellow solid.

¹H NMR (400MHz, chloroform-d)₁)δ8.12(d，J＝9.2Hz，2H)，6.60(d，J＝9.2Hz，2H)，4.45(t，J＝5.6Hz，2H)，3.63(t，J＝5.6Hz，2H)，3.06(s，3H)

LCMS：183(M+H⁺)，C₈H₁₀N₂O₃S

<5-2>Preparation of Compound 19

Compound 18(22g, 82.00mmol), hydroxyphthalimide (17.4g, 107.04mmol) and triethylamine (17.3mL, 123.5mmol) were added to acetonitrile (300mL), then stirred and refluxed for 6 hours. Upon completion of the reaction, dichloromethane (1000mL) was added to the reaction solution. The reaction was washed with 0.5N-HCl solution (500mL) and saturated sodium bicarbonate solution (500mL) and then concentrated under reduced pressure to give compound 19(26g, 79.44mmol, 97%) as a yellow solid.

¹H NMR (400MHz, chloroform-d)₁)δ8.12(d，J＝9.2Hz，2H)，7.88(m，2H)，7.87(m，2H)，6.65(d，J＝9.2Hz，2H)，5.83(br s，1H)，4.45(t，J＝4.8Hz，2H)，3.56(m，2H)，

LCMS：328(M+H⁺)，C₁₆H₁₃N₃O₅

<5-3>Preparation of Compound 20

Compound 19(79.44mmol) and hydrazine (20mL) were added to ethanol, followed by stirring and refluxing for 2 hours. The reaction was cooled at room temperature, filtered, and then concentrated under reduced pressure to give compound 20(19g, crude). This compound was not purified and used for the next reaction.

¹H NMR(400MHz，DMSO-d₆)δ7.94(d，J＝9.6Hz，2H)，7.34(t，J＝5.6Hz，1H)，6.69(d，J＝9.5Hz，2H)，3.68(t，J＝5.2Hz，2H)，3.37(m，2H)

LCMS：198(M+H⁺)，C₈H₁₁N₃O₃

<5-4>Preparation of Compound 21

Compound 20(19g) and sodium carbonate (21g, 198mmol) were dissolved in dioxane (200mL) and distilled water (200mL), and di-tert-butoxycarbonyl (ditertbutoxycarbonyl) (26g, 119mmol) was slowly added thereto, followed by stirring at room temperature for 4 hours. Upon completion of the reaction, the reaction was filtered under reduced pressure, washed with distilled water (1000mL) and dried to give compound 21(26g, crude) as a yellow solid.

¹H NMR (400MHz, chloroform-d)₁)δ8.10(d，J＝9.2Hz，2H)，6.54(d，J＝9.2Hz，2H)，5.50(s，1H)，5.04(s，1H)，3.91(t，J＝5.2Hz，2H)，3.45(m，2H)，1.43(s，9H)

LCMS：298(M+H⁺)，C₁₃H₁₉N₃O₅

<5-5>Preparation of Compound 22

Compound 21(13g, 40mmol), acetic acid (132mL) and concentrated HCl (10mL, 300mmol) were dissolved in tetrahydrofuran (200mL), to which zinc (26g, 400mmol) was added at 0 deg.C, followed by stirring for 2 hours. A20% ammonia solution (200mL) was added slowly at 0 deg.C, followed by dichloromethane (500 mL). The organic layer was separated and then concentrated under reduced pressure to give compound 22(6.5g, 24.31mmol, 61%) as a white solid.

¹H NMR (400MHz, chloroform-d)₁)δ7.17(s，1H)，6.60(d，J＝8.4Hz，2H)，6.60(d，J＝8.4Hz，2H)，4.03(t，J＝4.8Hz，2H)，3.30(t，J＝4.8Hz，2H)，1.47(s，9H)

LCMS：268(M+H⁺)，C₁₃H₂₁N₃O₃

<5-6>Preparation of Compound 23

Compound 22(6.5g, 24.31mmol) and (S) -glycidylphthalimide (3.95g, 19.45mmol) were added to isopropanol (100mL), followed by stirring and refluxing for 6 hours. Upon completion of the reaction, the reaction was concentrated under reduced pressure and then subjected to column chromatography to give compound 23(9g, crude product) as a white solid.

¹H NMR (400MHz, chloroform-d)₁)δ7.85(m，2H)，7.79(m，2H)，6.33(d，J＝9.2Hz，2H)，6.21(d，J＝9.2Hz，2H)，4.07(m，3H)，3.58-3.31(m，6H)，1.46(s，9H)

LCMS：471(M+H⁺)，C₂₄H₃₀N₄O₆

<5-7>Preparation of Compound 24

Compound 23(9g, crude) and carbonyldiimidazole (3.5g, 29.16mmol) were added to tetrahydrofuran (150mL), followed by stirring for 12 hours. On completion of the reaction, the reaction was concentrated under reduced pressure and then subjected to column chromatography to give compound 24(1.6g, 3.22mmol) as a white solid.

¹H NMR (400MHz, chloroform-d)₁)δ7.88(m，2H)，7.75(m，2H)，7.28(d，J＝8.8Hz，2H)，6.55(d，J＝8.8Hz，2H)，4.95(m，1H)，4.17-3.82(m，6H)，3.37(t，J＝5.2Hz，2H)，1.46(s，9H)

LCMS：497(M+H⁺)，C₂₅H₂₈N₄O₇

<5-8>Preparation of Compound 25

Compound 24(1.6g, 3.22mmol) and hydrazine (1.6mL, 32.20mmol) were added to ethanol (30mL), then stirred and refluxed for 2 hours. The reaction was cooled at room temperature, filtered, and then concentrated under reduced pressure to give compound 25(1.3g, crude). This compound was used in the next reaction without purification.

¹H NMR (400MHz, chloroform-d)₁)δ7.33(d，J＝9.2Hz，2H)，6.66(d，J＝9.2Hz，2H)，4.63(m，1H)，4.03(m，3H)，3.77(dd，J＝6.42.0Hz，1H)，3.34(t，J＝5.2Hz，2H)，3.03(m，2H)，1.49(s，9H)

LCMS：367(M+H⁺)，C₁₇H₂₆N₄O₅

<5-9>Preparation of Compound 26

Compound 25(1.3g, 3.22mmol), 5-chlorothiophene-2-carboxylic acid (0.68g, 4.19mmol) and PyBOP [ (benzotriazol-1-yloxy) tripyrrolidinylphosphonium hexafluorophosphate ] (2.5g, 4.83mmol) were added to N, N-dimethylformamide (20mL), to which diisopropylethylamine (1.06mL, 6.44mmol) was slowly added at 0 ℃ and then stirred for 1 hour. When the reaction was complete, ethyl acetate (300mL) was added to the reaction solution. The reaction was washed 2 times with distilled water (200mL), then concentrated under reduced pressure to give compound 26 as a white solid (1.6g, 3.13 mmol).

¹H NMR(400MHz，DMSO-d₆)δ10.05(s，1H)，8.97(t，J＝5.6Hz，1H)，7.70(d，J＝4.0Hz，1H)，7.20(m，3H)，6.60(d，J＝8.4Hz，2H)，4.76(m，1H)，4.07(t，J＝8.8Hz，1H)，3.83(t，J＝5.6Hz，2H)，3.74(m，1H)，3.57(t，J＝5.2Hz，2H)，3.21(t，J＝5.6Hz，2H)，1.40(s，9H)

LCMS：511(M+H⁺)，C₂₂H₂₇ClN₄O₆S

Example 1: preparation of Compound 100

Compound 5(5.0g, 13.8mmol) obtained in preparation example 1 was added to anhydrous methanol (200mL), followed by bubbling of HCl gas at 0 ℃ for 30 minutes. Methanol (100mL) was added thereto, followed by an additional 30 minutes of HCl gas, followed by stirring at room temperature for 2 hours. The reaction was concentrated under reduced pressure to remove the remaining HCl. To this was added acetic acid (120mL) and 2- (N-methylamino) ethyl-hydroxylamine hydrochloride (3.5g, 27.6mmol), followed by overnight reflux. The reaction solution was concentrated under reduced pressure, dissolved in ethyl acetate and then saturated NaHCO₃The solution was washed and then column chromatographed to give the title compound 100 as a white solid (1.85g, 4.3mmol, 31%).

¹H NMR(400MHz，CDCl₃)δ7.54(d，J＝9.0Hz，2H)，7.44(d，J＝9.0Hz，2H)，7.34(d，J＝4.2Hz，1H)，6.89(br t，1H)，4.80(m，1H)，4.12(t，J＝4.8Hz，2H)，4.04(t，J＝9.0Hz，1H)，3.85-3.80(m，2H)，3.69-3.64(m，1H)，3.45(t，J＝4.8Hz，2H)，2.76(s，3H)

LCMS：435(M+H⁺)，C₁₉H₁₉ClN₄O₄S

Example 2: preparation of Compound 101

Compound 101 was prepared as a white solid (17mg, 0.04mmol, 14%) in the same manner as described in example 1 using compound 5(0.1g, 0.27mmol) obtained in preparation example 1 and (aminoethyl) hydroxylamine (62mg, 0.8 mmol).

¹H NMR(400MHz，DMSO-d₆)δ8.98(t，J＝6.0Hz，1H)，7.68(d，J＝4.0Hz，1H)，7.63(d，J＝8.8Hz，2H)，7.57(d，J＝8.8Hz，2H)，7.19(d，J＝4.0Hz，1H)，7.07(s，1H)，4.85(m，1H)，4.19(t，J＝8.8Hz，1H)，3.86-3.81(m，3H)，3.61(t，J＝5.2Hz，2H)，3.38(m，2H)

LCMS：421(M+H⁺)，C₁₈H₁₇ClN₄O₄S

Example 3: preparation of Compound 102

Compound 102 was prepared as a white solid (30mg, 48%) in the same manner as described in example 1 using compound 5(100mg, 0.27mmol) obtained in preparation example 1 and O- [2- (2-ethylamino) -ethyl ] -hydroxylamine (15mg, 0.139 mmol).

¹H NMR(400MHz，CDCl₃)δ7.52(d，J＝8.4Hz，2H)，7.42(d，J＝8.4Hz，2H)，7.37(d，J＝3.6Hz，1H)，7.28-7.22(m，1H)，6.85(d，J＝3.6Hz，1H)，4.82-4.71(m，1H)，4.11(t，J＝4.4Hz，2H)，3.98(t，J＝8.8Hz，1H)，3.83-3.72(m，2H)，3.66-3.56(m，1H)，3.43(t，J＝4.4Hz，2H)，3.01(q，J＝7.2Hz，2H)，1.03(t，J＝4.4Hz，3H)

LCMS：449(M+H⁺)，C₂₀H₂₁ClN₄O₄S

Example 4: preparation of Compound 103

Compound 103 was prepared as a white solid (31mg, 29%) in the same manner as described in example 1 using compound 5(100mg, 0.27mmol) obtained in preparation example 1 and O- [2- (2-cyclopropylamino) -ethyl ] -hydroxylamine (27mg, 0.23 mmol).

¹H NMR(400MHz，CDCl₃)δ7.52(d，J＝7.6Hz，2H)，7.46(d，J＝7.6Hz，2H)，7.38(d，J＝3.6Hz，1H)，7.23-7.18(m，1H)，6.87(d，J＝3.6Hz，1H)，4.85-4.72(m，1H)，4.06(t，J＝4.4Hz，2H)，4.02(t，J＝8.4Hz，1H)，3.85-3.74(m，2H)，3.68-3.55(m，1H)，3.53(t，J＝4.4Hz，2H)，2.62-2.51(m，1H)，0.51-0.32(m，4H)

LCMS：461(M+H⁺)，C₂₁H₂₁ClN₄O₄S

Example 5: preparation of Compound 104

Compound 104 was prepared as a white solid (63.7mg, 0.146mmol, 58%) in the same manner as described in example 1 using compound 5 obtained in preparation example 1 (100mg, 0.27mmol) and N-methyl-O- (2-aminoethyl) hydroxylamine dihydrochloride (180.0mg, 1.104 mmol).

¹H NMR(400MHz，DMSO-d₆)δ9.07(t，J＝6.4Hz，1H)，7.70(d，J＝4.4Hz，1H)，7.54(m，4H)，7.14(d，J＝4.4Hz，1H)，4.80(m，1H)，4.15(t，J＝8.8Hz，1H)，3.86(dd，J＝8.8Hz，8.8Hz，1H)，3.81(t，J＝4.8Hz，2H)，3.55-3.48(m，4H)，2.82(s，3H)。

LCMS：435(M+H⁺)，C₁₉H₁₉ClN₄O₄S

Example 6: preparation of Compound 105

Compound 105 was prepared as a pale yellow solid (9.1mg, 0.020mmol, 9%) in the same manner as described in example 1 using compound 5 obtained in preparation example 1 (80.0g, 0.221mmol) and N-ethyl-O- (2-aminoethyl) hydroxylamine dihydrochloride (300.0mg, 1.69 mmol).

¹H NMR(600MHz，DMSO-d₆)δ9.37(br，1H)，7.88(m，1H)，7.79(d，J＝6.0Hz，2H)，7.73(d，J＝5.6Hz，2H)，7.20(d，J＝4.2Hz，1H)，4.91(m，1H)，4.24(t，J＝8.7Hz，1H)，4.05(dd，J＝8.7Hz，8.7Hz，1H)，3.66(t，J＝4.8Hz，2H)，3.63-3.54(m，4H)，2.81(q，J＝7.2Hz，2H)，1.24(t，J＝7.2Hz，3H)

LCMS：449(M+H⁺)，C₂₀H₂₁ClN₄O₄S

Example 7: preparation of Compound 106

Compound 106 was prepared as a white solid (20mg, 0.04mmol, 16%) in the same manner as described in example 1 using compound 5 obtained in preparation example 1 (100mg, 0.27mmol) and 2- (1-methylhydrazino) -N-ethylethylamine (93mg, 0.8 mmol).

¹H NMR(600MHz，DMSO-d₆) δ 8.99(t, J ═ 5.4Hz, 1H), 7.69(d, J ═ 3.6Hz, 1H), 7.53(d, J ═ 8.4Hz, 2H), 7.36(t, J ═ 8.4Hz, 2H), 7.19(d, J ═ 3.6Hz, 1H), 4.84(m, 1H), 4.19(t, J ═ 9.6Hz, 1H), 3.86(dd, J ═ 8.46.6Hz, 1H), 3.61(t, J ═ 5.4Hz, 2H), 3.31(m, 2H), 2.92(m, 2H), 2.80 (brs, 2H), 2.59(s, 3H), 0.94(t, J ═ 7.2, 1H) (according to acetic acid equivalent, 2H)

LCMS：462(M+H⁺)，C₂₁H₂₄ClN₅O₃S

Example 8: preparation of Compound 107

NaH (3.3g, 87.01mmol) was added to N, N-dimethylformamide (30mL) and then stirred for 15 min. Tert-butyl carbazate (Tertbutylchabazite) (5g, 37.83mmol) was dissolved in N, N-dimethylformamide (10mL) and added slowly thereto at 0 ℃. Dibromopropane (7.6g, 37.83mmol) was also added thereto at 0 ℃ and then stirred at room temperature for 3 hours. The reaction was cooled at 0 ℃ and washed with distilled water (50 mL). When the reaction was complete, the reaction was dissolved in ethyl acetate (500mL) and washed 3 times with sodium bicarbonate solution (50 mL). Column chromatography gave compound 107a as an oil (2.4g, 13.93mmol, 36.7%).

¹H NMR (400MHz, chloroform-d)₁)δ3.85(s，1H)，3.45(t，J＝7.2Hz，2H)，3.04(t，J＝6.4Hz，2H)，2.03(m，2H)，1.49(s，9H)

LCMS：173(M+H⁺)，C₈H₁₆N₂O₂

Compound 107a (1.5g, 8.71mmol), N-2-bromophthalimide (2.33g, 8.71mmol) and potassium carbonate (1.32g, 9.58mmol) were added to N, N-dimethylformamide (10mL), followed by stirring at 100 ℃ for 12 hours. The reaction was dissolved in ethyl acetate (200mL) and washed 3 times with sodium bicarbonate solution (30 mL). Column chromatography gave compound 107b as an oil (1.7g, 4.92mmol, 56.2%).

¹H NMR (400MHz, chloroform-d)₁)δ7.89(m，2H)，7.74(m，2H)，3.94(tJ＝4.4Hz，2H)，3.90(t，J＝5.2Hz，2H)，3.11(t，J＝6.0Hz，2H)，3.06(t，J＝4.4Hz，2H)，2.01(m，2H)，1.48(s，9H)，

LCMS：346(M+H⁺)，C₁₈H₂₃N₃O₄

Compound 107b (0.5g, 1.45mmol) was added to 4M-HCl (in dioxane) (5mL) and stirred for 0.5 h. The reaction mixture was concentrated under reduced pressure, to which methanol (5mL) and methylamine (4mL) were added, followed by stirring and refluxing for 1 hour. Concentration of the reaction under reduced pressure afforded compound 107c, which was used without further purification for the next reaction.

LCMS：116(M+H⁺)，C₅H₁₃N₃

Compound 5(100mg, 0.27mmol) obtained in preparation example 1 was added to anhydrous methanol (10mL), followed by introduction of HCl gas at 0 ℃ for 30 minutes and stirring at room temperature for 2 hours. The reaction was concentrated under reduced pressure to remove the remaining HCl. To this were added anhydrous methanol (10mL) and compound 107c (300mg, 0.8mmol) stepwise, followed by stirring at room temperature for 12 hours, followed by concentration under reduced pressure. Thereafter, preparative TLC treatment afforded title compound 107 as a white solid (60mg, 0.13mmol, 48%).

¹H NMR(400MHz，DMSO-d₆)δ9.12(t J＝5.6Hz，1H)，7.77-7.64(m，5H)，7.14(d，J＝4.4Hz，1H)，4.84(m，1H)，4.18(t，J＝8.8Hz，1H)，3.90(dd J＝8.8，6.0Hz，1H)，3.70(t，J＝6.8Hz，1H)，3.65(t，J＝6.8Hz，1H)，3.59-3.52(m，3H)，3.25(m，1H)，3.11(m，1H)，2.95(m，1H)，2.65(m，1H)，2.55(m，1H)，2.08(m，2H)

LCMS：460(M+H⁺)，C₂₁H₂₂ClN₅O₃S

Example 9: preparation of Compound 108

Tert-butyl carbazate (2.5g, 18.91mmol), N- (bromoethyl) phthalimide (5.25g, 20.80mmol) and potassium carbonate (3.14g, 22.70mmol) were added to N, N-dimethylformamide (30mL), followed by stirring at 90 ℃ for 12 hours. When the reaction was complete, the reaction was dissolved in ethyl acetate (250mL) and washed 3 times with sodium bicarbonate solution (150 mL). After concentration under reduced pressure, column chromatography gave compound 108a as a white solid (1g, 3.2mmol, 19%).

¹H NMR (400MHz, chloroform-d)₁)δ7.85(m，2H)，7.73(m，2H)，6.49(s，1H)，4.12(s，1H)，3.83(t，J＝3.6Hz，2H)，3.05(m，2H)，1.46(s，9H)，

LCMS：306(M+H⁺)，C₁₅H₁₉N₃O₄

Compound 108a (0.9g, 2.95mmol) and potassium carbonate (1.03g, 7.4mmol) were dissolved in N, N-dimethylformamide (10mL), 3-bromopropionyl chloride (0.7g, 3.6mmol) was added thereto, followed by stirring at 90 ℃ for 5 hours. Ethyl acetate (100mL) was added and washed with sodium bicarbonate solution (30 mL). After concentration under reduced pressure, column chromatography gave compound 108b as an oil (230mg, 0.64mmol, 22%).

¹H NMR (400MHz, chloroform-d)₁)δ7.85(m，2H)，7.73(m，2H)，6.49(s，1H)，4.12(s，1H)，3.83(t，J＝3.6Hz，2H)，3.05(m，2H)，1.46(s，9H)，

LCMS：360(M+H⁺)，C₁₈H₂₁N₃O₅

Compound 108b (230mg, 0.64mmol) was added to 4M-HCl (in dioxane) (2mL) and stirred for 1 h. The reaction was concentrated under reduced pressure, and methanol (5mL) and methylamine (2mL) were added thereto, followed by stirring for 1 hour. Concentration of the reaction under reduced pressure afforded compound 108c (103mg, 0.64mmol), which was used in the next reaction without further purification.

LCMS：130(M+H⁺)，C₅H₁₁N₃O

Compound 108 was prepared as a white solid (26mg, 0.05mmol, 19%) in the same manner as described in example 1 using compound 5(0.1g, 0.27mmol) obtained in preparation example 1 and compound 108c (103mg, 0.64 mmol).

¹H NMR(600MHz，DMSO-d₆)δ9.09(t，J＝4.8Hz，1H)，7.79(d，J＝8.4Hz，2H)，7.72-7.71(m，3H)，7.19(d，J＝4.2Hz，1H)，4.89(m，1H)，4.24(t，J＝9.0Hz，1H)，4.12(t，J＝7.8Hz，2H)，3.94-3.89(m，3H)，3.62(m，2H)，3.53(t，J＝4.8Hz，2H)，2.80(t，J＝8.4Hz，2H)，3.59-3.52(m，3H)，3.25(m，1H)，3.11(m，1H)，2.95(m，1H)，2.65(m，1H)，2.55(m，1H)，2.08(m，2H)

LCMS：474(M+H⁺)，C₂₁H₂₀ClN₅O₄S

Example 10: preparation of Compound 109

Compound 15a (450mg, 0.88mmol) obtained in preparation example 3 was dissolved in dichloromethane (10mL), to which HCl (4M 1, 4-dioxane solution) (10mL) was added, followed by stirring at room temperature for 1 hour. The reaction was concentrated under reduced pressure, then dried to give a pale yellow solid compound (425mg, 0.88mmol, 100%). This compound (392mg, 0.81mmol) was dissolved in acetic acid (4mL), to which was added trimethyl orthoformate (2mL), followed by stirring and refluxing. After 10 h, all solvents were evaporated and then treated by column chromatography (dichloromethane/methanol (v/v)20/1 → 12/1) to give the title compound 109 as a pale yellow solid (215mg, 5.12mmol, 63%).

¹H NMR(400MHz，CDCl₃)δ7.35(d，J＝9.2Hz，2H)，7.33(d，J＝4.4Hz，1H)，7.14(d，J＝9.2Hz，2H)，7.01(t，J＝6.4Hz，1H)，6.88(s，1H)，6.85(d，J＝4.4Hz，1H)，4.87-4.79(m，1H)，4.06(t，J＝9Hz，1H)，3.86(ddd，J＝14.4，6，3Hz，1H)，3.81(dd，J＝9，6.4Hz，1H)，3.69(dt，J＝14.4，6Hz，1H)，3.62-3.58(m，2H)，3.55-3.51(m，2H)

LCMS：420(M+H⁺)，C₁₈H₁₈ClN₅O₃S

Example 11: preparation of Compound 110

The title compound 110 was obtained in a similar manner to that described in example 10 using triethyl orthoacetate instead of trimethyl orthoformate of example 10 using compound 15a obtained in preparation example 3.

¹H NMR(600MHz，CDCl₃)δ7.36(d，J＝9Hz，2H)，7.30(d，J＝4Hz，1H)，7.17(d，J＝9Hz，2H)，6.90(d，J＝4.2Hz，1H)，6.49(br t，1H)，4.85-4.80(m，1H)，4.28(br，1H)，4.08(t，J＝9Hz，1H)，3.95-3.90(m，1H)，3.79(t，J＝7.8Hz，1H)，3.72-3.67(m，1H)，3.62-3.57(m，2H)，3.49-3.44(m，2H)，1.98(s，3H)

LCMS：434(M+H⁺)，C₁₉H₂₀ClN₅O₃S

Example 12: preparation of Compound 111

The title compound 111 was obtained in a similar manner to that described in example 10 using triethyl orthopropionate instead of trimethyl orthoformate of example 10 using compound 15a obtained in preparation example 3.

¹H NMR(600MHz，CDCl₃)δ7.35(d，J＝9Hz，2H)，7.30(d，J＝4.2Hz，1H)，7.18(d，J＝9Hz，2H)，6.90(d，J＝4.2Hz，1H)，6.53(br t，1H)，4.85-4.80(m，1H)，4.27(br，1H)，4.08(t，J＝9Hz，1H)，3.92(ddd，J＝14.4，6.6，3Hz，1H)，3.79(t，J＝7.8Hz，1H)，3.72-3.67(m，1H)，3.62-3.57(m，2H)，3.50-3.44(m，2H)，2.26(q，J＝7.8Hz，2H)，1.20(t，J＝7.8Hz，3H)

LCMS：448(M+H⁺)，C₂₀H₂₂ClN₅O₃S

Example 13: preparation of Compound 112

The title compound 112 was obtained in a similar manner to that described in example 10 using compound 15b obtained in preparation example 4.

¹H NMR(600MHz，CDCl₃)δ7.35(d，J＝9Hz，2H)，7.33(d，J＝4.5Hz，1H)，7.13(d，J＝9Hz，2H)，6.93(t，J＝6Hz，1H)，6.85(d，J＝4.5Hz，1H)，6.64(s，1H)，4.86-4.80(m，1H)，4.06(t，J＝9Hz，1H)，3.91-3.85(m，1H)，3.80(dd，J＝9，7Hz，1H)，3.71-3.65(m，1H)，3.54(t，J＝4.8Hz，2H)，3.43(t，J＝4.8Hz，2H)，2.90(s，3H)

LCMS：434(M+H⁺)，C₁₉H₂₀ClN₅O₃S

Example 14: chemical combinationPreparation of substance 113

The title compound 113 was obtained in a similar manner to that described in example 10 using triethyl orthoacetate instead of trimethyl orthoformate of example 10 using compound 15b obtained in preparation example 4.

¹H NMR(600MHz，CDCl₃)δ7.34(d，J＝9Hz，2H)，7.31(d，J＝4Hz，1H)，7.15(d，J＝9Hz，2H)，6.88(d，J＝4Hz，1H)，6.69(t，J＝6Hz，1H)，4.84-4.78(m，1H)，4.06(t，J＝9Hz，1H)，3.90(ddd，J＝11，7，3Hz，1H)，3.79(dd，J＝9，6.6Hz，1H)，3.68(dt，J＝14.4，6.6Hz，1H)，3.52(t，J＝5Hz，2H)，3.44(t，J＝5Hz，2H)，2.93(s，3H)，2.05(s，3H)

LCMS：448(M+H⁺)，C₂₀H₂₂ClN₅O₃S

Example 15: preparation of Compound 114

In a similar manner to that described in example 10, using compound 16a obtained in preparation example 3, the title compound 114 was obtained as a white solid (7.4mg, 0.014mmol, 47%).

¹H NMR(600MHz，DMSO-d₆)δ9.03(t，J＝5.4Hz，1H)，8.65(s，1H)，7.68(d，J＝3.6Hz，1H)，7.55(d，J＝8.4Hz，2H)，7.19(d，J＝3.6Hz，1H)，7.14(d，J＝8.4Hz，2H)，4.88-4.81(m，1H)，4.17(t，J＝9Hz，1H)，3.85-3.81(m，1H)，3.70-3.50(m，4H)，3.29(s，3H)，3.13-3.05(m，2H)

LCMS：434(M+H⁺)，C₁₉H₂₀ClN₅O₃S

Example 16: preparation of Compound 115

Compound 26 obtained in preparation example 5 was added to 4M HCl (2mL), followed by stirring for 1 hour. The reaction was concentrated under reduced pressure, to which was added trimethyl orthoformate (2mL) and acetic acid (4mL), followed by stirring and refluxing for 12 hours. This reaction was subjected to column chromatography to give the title compound as a white solid (11mg, 0.03mmol, 8%).

¹H NMR (400MHz, chloroform-d)₁)δ7.55(s，1H)，7.50(d，J＝9.2Hz，2H)，7.42(br s，1H)，7.39(d，J＝4.0Hz，1H)，7.03(d，J＝9.2Hz，2H)，6.89(d，J＝4.0Hz，1H)，4.86(m，1H)，4.20(t，J＝4.8Hz，2H)，4.12(m，1H)，3.87(m，1H)，3.83-3.74(m，4H)

LCMS：421(M+H⁺)，C₁₈H₁₇ClN₄O₄S

Example 17: preparation of Compound 116

Compound 116 was prepared as a white solid in a similar manner to that described in example 14 using compound 26 obtained in preparation example 5.

¹H NMR(400MHz，DMSO-d₆)δ8.93(t，J＝5.6Hz，1H)，7.63(d，J＝4.2Hz，1H)，7.50(d，J＝8.8Hz，2H)，7.25(d，J＝8.8Hz，2H)，7.14(d，J＝4.2Hz，1H)，4.79(m，1H)，4.13(t，J＝8.8Hz，1H)，3.94(t，J＝4.6Hz，2H)，3.793.56-3.51(m，4H)，1.56(s，3H)

LCMS：435(M+H⁺)，C₁₉H₁₉ClN₄O₄S

Example 18: preparation of Compound 117

Compound 26(0.20mg, 0.39mmol) obtained in preparation example 5, cyclopropylcarbonyl chloride (50mg, 0.47mmol), pyridine (61mg, 0.78mmol) and 4-dimethylaminopyridine (5mg) were added to dichloromethane (5mL), followed by stirring for 2 hours. When the reaction was completed, methylene chloride (50mL) was added thereto, followed by washing with distilled water (10mL) 2 times. After concentration under reduced pressure, column chromatography gave the amide compound as a white solid (150mg, 0.29mmol, 74%).

¹H NMR(400MHz，DMSO-d₆)δ9.90(s，1H)，8.98(t，J＝6.0Hz，1H)，7.69(d，J＝4.0Hz，1H)，7.62(d，J＝8.8Hz，2H)，7.42(d，J＝8.8Hz，2H)，7.19(d，J＝4.0Hz，1H)，4.84(m，1H)，4.21(t，J＝8.8Hz)，3.87(dd，J＝9.2，6.4Hz，1H)，3.78-3.74(m，4H)，3.61(t，J＝5.6Hz，2H)，1.22(m，1H)，0.78(m，2H)，0.59(m，2H)

The compound (0.15g, 0.29mmol) obtained above was added to 4N HCl (in dioxane) (2mL), followed by stirring at room temperature for 1 hour. The reaction was concentrated under reduced pressure. Toluene (5mL) and phosphorus oxychloride (45mg, 0.29mmol) were added, followed by stirring and reflux for 12 hours. Column chromatography afforded the title compound 117 as a white solid (11mg, 0.03mmol, 10%).

¹H NMR(400MHz, chloroform-d)₁)δ7.54(d，J＝8.8Hz，2H)，7.31(d，J＝4.4Hz，1H)，7.25(d，J＝8.8Hz，2H)，6.90(d，J＝4.4Hz，1H)，6.55(t，J＝4.8Hz，1H)，4.88(m，1H)，4.10(t，J＝4.8Hz，2H)，3.89(m，2H)，3.79(m，1H)，3.67(t，J＝4.8Hz，2H)，1.06(m，1H)，0.93(m，2H)，0.58(m，2H)

LCMS：461(M+H⁺)，C₂₁H₂₁ClN₄O₄S

Example 19: preparation of Compound 118

Compound 118 was prepared as a white solid in a similar manner to that described in example 10 using compound 33 synthesized according to the method of reaction formula 5.

¹H NMR (400MHz, chloroform-d)₁)δ8.55(s，1H)，7.52(d，J＝8.8Hz，2H)，7.33(d，J＝4.4Hz，1H)，7.10(s，1H)，7.05(d，J＝8.8Hz，2H)，6.89(d，J＝4.4Hz，1H)，6.76(t，J＝4.8Hz，1H)，4.88(m，1H)，4.11(t，J＝8.8Hz，1H)，4.00(t，J＝4.8Hz，2H)，3.91-3.77(m，3H)，3.74(t，J＝4.8Hz，2H)

LCMS：448(M+H⁺)，C₁₉H₁₈ClN₅O₄S

Example 20: preparation of Compound 119

Compound 119 was prepared as a white solid in a similar manner to that described in example 14, using compound 33 synthesized according to the method of reaction formula 5.

¹H NMR(400MHz，DMSO-d₆)δ8.94(br t，1H)，8.35(s，1H)，7.64(d，J＝4.4Hz，1H)，7.52(d，J＝8.8Hz，2H)，7.26(d，J＝8.8Hz，2H)，7.14(d，J＝4.4Hz，1H)，4.80(m，1H)，4.15(t，J＝8.4Hz，1H)，3.81-3.75(m，3H)，3.58-3.51(m，4H)，1.75(s，3H)

LCMS：462(M+H⁺)，C₂₀H₂₀ClN₅O₄S

Example 21: preparation of Compound 120

Compound 119 synthesized in example 20 was dissolved in methanol and then deformylated with HCl to give the title compound 120 as a white solid.

¹H NMR(400MHz，DMSO-d₆)δ8.98(br t，1H)，7.69(d，J＝4.4Hz，1H)，7.52(d，J＝8.8Hz，2H)，7.23-7.19(m，3H)，4.83(m，1H)，4.18(t，J＝8.4Hz，1H)，3.84(m，1H)，3.61-3.55(m，4H)，3.06(t，J＝4.8Hz，2H)，1.64(s，3H)

LCMS：434(M+H⁺)，C₁₉H₂₀ClN₅O₃S

Example 22: preparation of Compound 121

Compound 120 synthesized in example 21 was reacted with methyl iodide to give the title compound 121 as a white solid.

¹H NMR(400MHz，DMSO-d₆)δ8.98(br t，1H)，7.69(d，J＝4.4Hz，1H)，7.55(d，J＝8.8Hz，2H)，7.28(d，J＝8.8Hz，2H)，7.19(d，J＝4.4Hz，1H)，4.84(m，1H)，4.18(t，J＝8.4Hz，1H)，3.84(m，1H)，3.65-2.92(m，6H)，2.60(s，3H)，1.67(s，3H)

LCMS：448(M+H⁺)，C₂₀H₂₂ClN₅O₃S

Experimental example 1: inhibitory Activity of factor Xa (FXa) inhibitors

1) Reagents and materials

Chromogenic substrate S-2765 (N-Z-D-Arg-Gly-Arg-pNA.2HCl) necessary for determining factor Xa activity was purchased from Chromogenix. Human FXa was purchased from Enzyme Research Laboratories. 96-well microplates were purchased from Corning Life Sciences.

2) Inhibitory Activity of FXa inhibitor

The inhibitory activity of the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention against FXa was determined as follows.

The activity of the compounds on purified human FXa was determined in 96-well microplates at 37 ℃ using the chromogenic substrate S-2765 (Z-D-Arg-Gly-Arg-pNA.2HCl). Enzyme activity was determined in 100mM Tris-HCl buffer (pH 7.8) containing human FXa (2.6nM), NaCl (150mM), PEG 8000 (0.1%), test compound dilutions (1% DMSO) and S-2765(300 uM). The reaction was initiated by adding the substrate and absorbance was monitored continuously for 5min at 405nm using SpectraMax 190(Molecular Devices, USA). According to the Cheng-Prusoff equation (Ki ═ IC)₅₀/1+[S]K_m) Calculation of inhibition constant (K) for human FXa_i) In which [ S ]]Is the concentration of the substrate, K_mIs the Michalis-Menten constant. K_mDetermined from the Lineweaver-Burk plot. IC (integrated circuit)₅₀Is the amount of inhibitor required to reduce the initial rate of the control by 50%. Using GraFit software version 5.0.12 (version: (R))Erithocus Software Ltd., UK) to calculate IC₅₀The value is obtained.

The Km value used for the calculation was 125. mu.M, which was obtained by varying the substrate concentration at a constant enzyme concentration.

Experimental example 2: effect on blood coagulation

The effect of the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention on blood coagulation was investigated by measuring Prothrombin Time (PT).

1) PT assay

a) Method of using a coagulometer: prothrombin Time (PT) was measured using a Thrombovimer 4-channel hemagglutinometer (Behnk Elektronik, Germany). Citrate treated human and rat plasma were used in the assay. For the PT assay, 100uL of freshly thawed plasma was mixed with 3uL of serial dilutions of test compound or DMSO. After incubation at 37 ℃ for 5min, 200uL of STA-Neoplastine (diagnostic Stago, France) was added to initiate clot formation. The anticoagulant activity of the compounds is defined as doubling the plasma clotting time [ 2XPT (uM)]The desired concentration. Human plasma was obtained from the Daejeon Red Cross Blood Center. Rat blood was drawn from the carotid artery or superior vena cava under anesthesia. The blood was collected in plastic tubes containing 1/10 volumes of 3.8% sodium citrate. Plasma was obtained by direct centrifugation at 2500g for 10min at 4 ℃ and stored at-70 ℃.

b) Method using spectramax:according to the invention, serial dilutions of the compound (5uL) were mixed with citrate treated plasma (45uL) and after 5min at 37 ℃ STA-Neoplastine (diagnostic Stago, France) was added. The absorbance at 340nm was continuously monitored, and PT was measured as time (sec) when the absorbance at 340nm reached 0.1. The anticoagulant activity of the compounds is defined as doubling the plasma clotting time [ 2XPT (uM)]The desired concentration. The anticoagulant activity of the compound is determinedMeaning to double the plasma clotting time [ 2XPT (uM)]The desired concentration. Human plasma was obtained from the Daejeon Red Cross Blood Center. Rat blood was drawn from the carotid artery or superior vena cava under anesthesia. The blood was collected in plastic tubes containing 1/10 volumes of 3.8% sodium citrate. Plasma was obtained by direct centrifugation at 2500g for 10min at 4 ℃ and stored at-70 ℃.

2) Determination of antithrombotic Effect Using an arteriovenous short (AV-short) model in rats

The antithrombotic effect of the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention was evaluated using Arteriovenous (AV) short circuit in rats. Fasted male Sprague-Dawley rats weighing 200-240g and aged about 7 weeks were anesthetized by intraperitoneal injection of urethane (1.25g/kg) or chloral hydrate. As described above, a mobile venous (AV) short circuit was performed in anesthetized rats with minor modifications [ Journal of thrombosis and Haemostasis (2004)3, 514].2 200-mm long saline-filled catheters (PE-50, Becton Dickinson, USA) were inserted into the left common carotid artery and the right jugular vein. Polyethylene tubes were passed through a polysiloxane tube (L/S) of g-mm length13 MasterFlex, USA) with 50-mm long silicone tubing (L/S) containing 75-mm long cotton threads16, MasterFlex, USA). The compound or vehicle was administered orally, and after 60min, the shunt was opened for 15 min. The cotton thread was then withdrawn and weighed. Excel 2003(Microsoft Windows)) Calculation of ED by Linear regression analysis₅₀The value is obtained.

3) Determination of Bleeding Time (BT) Using rat tail bleeding model

Fasted male Sprague-Dawley rats weighing 200-240g and aged about 7 weeks were anesthetized by intraperitoneal injection of pentobarbital-Na (60 mg/kg). The FXa inhibitor or vehicle was administered orally and after 60min, 2mm was transected from the tail tips of anesthetized rats and immersed vertically in saline at 37 ℃. The time until continuous bleeding stopped > 30s was determined, with a maximum observation time of 30min (longer bleeding time assigned 30 min).

The inhibition constants, anticoagulant effect (expressed in the form of 2 xPT) and antithrombotic effect (expressed in the form of% inhibition of thrombosis) in rat AV-short as determined by the above experiments for human FXa are shown in table 1. The effect of the compounds on rat tail transection bleeding time is summarized in table 2. Rivaroxaban, represented by formula a, was used as a comparative drug.

[ formula A ]

[ Table 1]

Inhibition constant against human FXa, anticoagulant effect (expressed in the form of 2 xPT) and antithrombotic effect in rat AV-short-circuit (expressed in the form of% inhibition of thrombosis) of the compound of formula I

113	29.34	0.62	-	12*
					114	2.86	0.50	-	5*
115	83.97	-	1.49	29
					116	23.22	3.51	0.71	74
117	101	-	2.2	-
					118	132.05	9.38	4.88	74
119	64.43	4.77	2.53	72

120	645.19	-	3.75	43
					121	95.60	2.90	0.54	51

^*Uratan is used as an anesthetic. For the asterisked compound, chloral hydrate was used.

As shown in table 1, the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention have Ki and PT values similar to those of the comparative drug rivaroxaban. In the rat AV-short model, the antithrombotic effect of the compounds is comparable to rivaroxaban, although minor changes in antithrombotic effect were observed depending on the anesthetic used. The most serious side effect of rivaroxaban is bleeding. Tail bleeding time was evaluated at various doses to investigate the bleeding effects of representative compounds represented by formula I. The effect of compounds 100, 109 and the comparative drug rivaroxaban on rat tail transection bleeding time is summarized in table 2.

Table 2 effect of compounds 100 and 109 on tail bleeding time in rats (n ═ 13)

In the rat tail bleeding model, compound 100 did not prolong bleeding time even at 10mg/kg compared to vehicle controls. On the other hand, rivaroxaban extended bleeding time by 3-4 times even at 1.25 mg/kg. Thus, the compound of formula I was confirmed to significantly reduce this side effect (bleeding). Furthermore, the compounds of formula I can be formulated in the form of salts using acids such as methanesulfonic acid or HCl, which can improve water solubility. The water solubility was determined by the following experiment.

Experimental example 3: determination of Water solubility

Water solubility of compounds 100 and 109 (representative oxazolidinone derivatives of formula I of the present invention containing cyclic amidoxime group (100) and cyclic amidrazone group (109)) in the form of hydrochloride or Methanesulfonate (MSA) was determined, and the results are shown in table 3. Rivaroxaban, represented by formula a, was used as a comparative drug.

TABLE 3 Water solubility of Compounds 100 and 109 in the form of the hydrochloride or mesylate salt

Compound (I)	Water solubility (mM)
		Rivaroxaban	＜0.05

100-HCl	12
		100-MSA	＞20
109-HCl	6
		109-MSA	＞20

As shown in table 3, the preparation of oxazolidinone derivatives containing cyclic amidoxime (100) or cyclic amidrazone group (109) of formula I of the present invention in the form of salts has great advantages, and thus it may have excellent water solubility, 200 times that of rivaroxaban, which is a control substance. This result indicates that the oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group of formula I of the present invention have high availability in the form of compositions for oral administration and injection.

As described above, the oxazolidinone derivative having a cyclic amidoxime or cyclic amidrazone group of formula I of the present invention shows almost no bleeding (one of the serious side effects of conventional drugs such as rivaroxaban), but has similar inhibitory activity to rivaroxaban and has excellent solubility, and thus, it has excellent utility as a composition for oral administration and injection.

INDUSTRIAL APPLICABILITY

The novel oxazolidinone derivative containing a cyclic amidoxime or cyclic amidrazone group represented by formula I of the present invention may be a very safe drug, which does not increase bleeding (a serious side effect of rivaroxaban used as a control substance in the present invention), and has higher solubility in water than rivaroxaban, and thus it may be easily developed as a composition for oral administration or injection.

It should be appreciated by those skilled in the art that the conception and specific embodiment disclosed may be readily utilized as a basis for modifying or designing other embodiments for carrying out the same purposes of the present invention. Those skilled in the art will also appreciate that such equivalent embodiments do not depart from the spirit and scope of the invention as set forth in the appended claims.

Claims (8)

1. Oxazolidinone derivatives containing a cyclic amidoxime or cyclic amidrazone group represented by formula I, prodrugs, hydrates, solvates, isomers, or pharmaceutically acceptable salts thereof,

[ formula I ]

Wherein the content of the first and second substances,

ring a is a residue selected from the group consisting of the following structures:

R₁to R₁₂Independently H, (C)₁-C₇) Alkyl, (C)₃-C₇) Cycloalkyl group, (C)₆-C₁₂) Aryl or (C)₄-C₁₂) Heteroaryl containing 1-4 heteroatoms selected from O, S and N, R₃And R₄Through the reaction of (C)₃-C₅) Alkylene groups which may be substituted by carbonyl groups and R₁To R₁₂The alkyl, cycloalkyl, aryl or heteroaryl of (A) may be selected from (C)₁-C₇) Alkyl, halo (C)₁-C₇) Alkyl, (C)₁-C₇) One of alkoxy and halogen;

R₁₃is H, (C)₁-C₇) Alkyl, (C)₃-C₇) Cycloalkyl, formyl, (C)₁-C₇) Alkylcarbonyl group, (C)₁-C₇) Alkoxycarbonyl or (C)₆-C₁₂) And (4) an aryl group.

2. An oxazolidinone derivative, a prodrug, hydrate, solvate, isomer, or pharmaceutically acceptable salt thereof according to claim 1 wherein the oxazolidinone derivative is selected from the group consisting of formula II to formula XI:

[ formula II ]

[ formula III ]

[ formula IV ]

[ formula V ]

[ formula VI ]

[ formula VII ]

[ formula VIII ]

[ formula IX ]

[ formula X ]

[ formula XI ]

Wherein the content of the first and second substances,

R₁to R₁₂Independently of each other, H and (C)₁-C₇) Alkyl or (C)₃-C₇) A cycloalkyl group; r₁₃Is H, (C)₁-C₇) Alkyl, (C)₃-C₇) Cycloalkyl, formyl or (C)₁-C₇) An alkylcarbonyl group; and m is an integer of 1 to 3.

3. An oxazolidinone derivative, its prodrug, hydrate, solvate, isomer or pharmaceutically acceptable salt thereof as claimed in claim 2 wherein R is₁To R₁₂Independently of one another, H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; r₁₃Is H, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, formyl or acetyl; and m is an integer of 1.

4. An oxazolidinone derivative, a prodrug, hydrate, solvate, isomer, or pharmaceutically acceptable salt thereof according to claim 3 wherein the oxazolidinone derivative is selected from the group consisting of:

5. an anticoagulant pharmaceutical composition comprising an oxazolidinone derivative, a prodrug, a hydrate, a solvate, an isomer, or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4.

6. An anticoagulant pharmaceutical composition comprising the oxazolidinone derivative, a prodrug, a hydrate, a solvate, an isomer, or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, for preventing or treating thrombosis, myocardial infarction, arteriosclerosis, inflammation, stroke, angina pectoris, restenosis, intermittent claudication, venous thrombosis, pulmonary embolism, arterial thrombosis, myocardial ischemia, or thromboembolism.

7. An anticoagulant pharmaceutical composition comprising the oxazolidinone derivative, its prodrug, hydrate, solvate, isomer, or pharmaceutically acceptable salt according to any one of claims 1 to 4, and a thrombolytic drug for preventing or treating coronary artery disease, cerebral artery disease, or peripheral artery disease.

8. An anticoagulant pharmaceutical composition comprising an oxazolidinone derivative, a prodrug, a hydrate, a solvate, an isomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4 for use in the extracorporeal preservation of blood, plasma or a blood product.