KR20030080810A - Selective thrombin inhibitors with an aminopyridoaryl group - Google Patents

Abstract

PURPOSE: Selective thrombin inhibitors having an aminopyridoaryl group are provided, which thrombin inhibitors have improved thrombin inhibiting activity, improves selectivity to thrombin and improved oral absorption rate. CONSTITUTION: Selective thrombin inhibitors represented by the formula 1, and pharmaceutically acceptable salts, prodrugs, hydrides, solvates and isomers thereof are provided, wherein Q is aminopyridoaryl; W is hydrogen, alkyl(C=1-8), diarylmethyl(C=6-10), arylalkyl(C=5-12), heteroaryl(C=4-10), heteroarylalkyl(C=5-11), dicycloalkylmethyl(C=3-6) or cycloalkyl(C=3-8); X and Y are independently selected from the group consisting of hydrogen, alkyl(C=1-8), cycloalkyl(C=3-8), alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkylaminocarbonylalkyl, monoalkylaminocarbonylalkyl, aminocarbonylalkyl, methanesulfonyl, and O-alkyloxalyl; and n is 0, 1 or 2.

Description

Selective thrombin inhibitors with an aminopyridoaryl group

The present invention relates to a compound represented by the following formula (1), and a composition for preventing blood clotting or treating various thrombosis containing the compound as an active ingredient.

Where

Q is an aminopyridoaryl group,

W is hydrogen, alkyl (C = 1-8), diarylmethyl (C = 6-10), arylalkyl (C = 5-12), heteroaryl (C = 4-10), heteroarylalkyl (C = 5-11), dicycloalkylmethyl (C = 3-6), or cycloalkyl (C = 3-8),

X and Y are each independently hydrogen, alkyl (C = 1-8), cycloalkyl (C = 3-8), alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkyl Aminocarbonylalkyl, monoalkylaminocarbonylalkyl, aminocarbonylalkyl, methanesulfonyl, aminosulfonyl, and O-alkyloxalyl,

n is 0, 1 or 2.

In general, it is known that various complex enzyme reactions are involved in the coagulation process. The final step involves the conversion of prothrombin to thrombin. Thrombin generated in this process activates platelets, converts fibrinogens to fibrin, and the like, and the fibrin is converted into a polymer by a polymerization reaction and crosslinked by activated blood factor XIII to become insoluble coagulation. Thrombin also plays a role in activating the blood factors V and VIII involved in the coagulation process, further accelerating the coagulation reaction. Therefore, the inhibitor of thrombin acts as an effective anticoagulant and at the same time inhibits platelet activity and prevents fibrin production and stabilization, thus developing a new substance that can inhibit thrombin activity for a long time to prevent blood clotting and prevent various thrombosis. Methods for treatment have been sought.

However, selectivity to inhibit thrombin only is essential for use as an effective anticoagulant. The reason for this is that there are various serine proteases having a thrombin-like active site structure in the human body, and the thrombin inhibitor lacking selectivity causes side effects such as bleeding. In general, the selectivity of a thrombin inhibitor is judged by comparing its ability to inhibit trypsin, a representative enzyme of serine protease. Under these circumstances, extensive research has been conducted to develop selective thrombin inhibitors that effectively inhibit thrombin and have low inhibitory activity against trypsin.

On the other hand, as the need for preventive anticoagulant treatment as well as the treatment of various diseases caused by blood clots, research and development is focused on the effort to develop an oral thrombin inhibitor that is easy to use in addition to the injection.

Representative compounds developed as effective thrombin inhibitors include, first, agatroban (Argatroban, dissociation constant (Ki) = 1.9 nM) of the following arylsulfonylarginine-based compounds (US Pat. No. 4,425,292 and US4201863).

[Formula A]

This compound has been reported to have a 250-fold thrombin inhibitory effect compared to trypsin and has already been commercialized as an injectable preparation in Japan in 1990 (Biochemistry 1984, 23, 85-90).

In addition, as a thrombin inhibitor, a benzamidine-based arylsulfonyl compound, NAPAP (Ki = 6.0 nM) of Formula B, was developed, which is easy to synthesize and exhibits a thrombin inhibitory effect compared to trypsin, despite being an effective thrombin inhibitor. The problem is that it is only about double (see J. Biol. Chem. 1991, 266, 20085-20093).

[Formula B]

Meanwhile, Ro46-6240 compound of formula C (Ki = 0.2 nM), which has improved selectivity compared to trypsin, has been reported as a potent thrombin inhibitor, which shows the possibility of development as an intravenous preparation, but has a short half-life in blood and is administered orally. There is no potential for development as a zero (see J. Med. Chem. 1994, 37, 3889-3901).

[Formula C]

A compound developed as an effective thrombin inhibitor capable of oral administration is CVS-1123 (Ki = 1.4 nM) of Formula D, developed by Corvas. It has been reported that this compound can be orally administered to dogs and monkeys as well as to mice, but has a disadvantage of poor selectivity for trypsin (up to 50 times; see WO 9315756 and WO 9408941).

[Formula D]

On the other hand, as a thrombin inhibitor developed by Akzo Nobel, the compound of formula E (IC ₅₀ = 34 nM) has a low selectivity for trypsin (IC ₅₀ = 4 nM), which is a major disadvantage and is not orally absorbed. Do not. (See WO 9730073 and 15th EFMC International Symposium on Medicinal Chemistry, Edinburgh, 1998, UK).

[Formula E]

Accordingly, the present inventors have intensively researched to develop new compounds having good thrombin inhibitory activity and ultimately excellent selectivity to thrombin compared to trypsin, and as a result, the compound of Formula 1 defined above has It was confirmed that this can be effectively achieved and came to complete the present invention.

Accordingly, it is an object of the present invention to provide a compound of formula 1, a pharmaceutically acceptable salt, prodrug, hydrate, solvate and isomer thereof.

Another object of the present invention for the prevention of blood coagulation and thrombosis containing a compound of Formula 1, pharmaceutically acceptable salts, prodrugs, hydrates, solvates and isomers as an active ingredient together with a pharmaceutically acceptable carrier. It is to provide a pharmaceutical composition.

Compounds excellent as thrombin inhibitors provided according to the invention are represented by the following general formula (1):

[Formula 1]

Where

Q is an aminopyridoaryl group,

W is hydrogen, alkyl (C = 1-8), diarylmethyl (C = 6-10), arylalkyl (C = 5-12), heteroaryl (C = 4-10), heteroarylalkyl (C = 5-11), dicycloalkylmethyl (C = 3-6), or cycloalkyl (C = 3-8),

X and Y are each independently hydrogen, alkyl (C = 1-8), cycloalkyl (C = 3-8), alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkyl Aminocarbonylalkyl, monoalkylaminocarbonylalkyl, aminocarbonylalkyl, methanesulfonyl, aminosulfonyl, and O-alkyloxalyl,

n is 0, 1 or 2.

Alkyl (C = 1-8) as used herein means straight or branched chain alkyl including methyl, ethyl, isopropyl, isobutyl, t-butyl, although the carbon number of the alkyl can be expected by those skilled in the art. It can be expanded to the extent that it is. Further, in the above substituent definition, arylalkyl, heteroarylalkyl, alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkylaminocarbonylalkyl, monoalkylaminocarbonialkyl, aminocarbonylalkyl Or in the O-alkyloxalyl the alkyl moiety has the same meaning as mentioned above.

The compound according to the present invention, in addition to the compound represented by Formula 1, includes a pharmaceutically acceptable salt thereof, a prodrug, a hydrate, a solvate and an isomer converted to the compound.

Pharmaceutically acceptable salts include acids that form non-toxic acid addition salts containing pharmaceutically acceptable anions, for example inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, and the like, tartaric acid, formic acid, citric acid Organic carbonic acid such as acetic acid, trichloroacetic acid or trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, etc., sulfonic acid such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid, and the like. Acid addition salts formed by

In particular, representative forms used as prodrugs include ester or amide compounds of this group if a carboxylic acid group is present in the compound of Formula 1, and N-alkylcarbonyl of this group if an amine group is present in the compound of = Amide), N-alkoxycarbonyl (= carbamate), N-hydroxy, N-alkoxy, N-alkylcarbonyloxy or N-oxide compound. In addition, the compounds according to the invention may have asymmetric carbon centers and therefore may exist as racemic compounds, diastereomeric mixtures and individual diastereoisomers, all of these isomeric forms being included within the scope of the invention.

Among the compounds of formula 1 according to the invention are preferred compounds having the following substituent definitions:

Q is an aminopyridoaryl group,

W is alkyl (C = 1-8), diarylmethyl (C = 6-10), arylalkyl (C = 5-12), heteroaryl (C = 4-10), heteroarylalkyl (C = 5--10 11) dicycloalkylmethyl (C = 3-6) or cycloalkyl (C = 3-8),

X is hydrogen, alkyl (C = 1-8), cycloalkyl (C = 3-8), alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkylaminocarbonylalkyl, Monoalkylaminocarbonylalkyl, aminocarbonylalkyl, methanesulfonyl, aminosulfonyl, and O-alkyloxalyl,

Y is hydrogen, alkyl (C = 1-8) or cycloalkyl (C = 3-8),

n is 0, 1 or 2.

Particularly preferred among the compounds of formula 1 according to the invention are those having the following substituent definitions:

Q is 1-amino-isoquinolin-6-yl, 1-amino-isoquinolin-7-yl, 1-amino-2-oxo-isoquinolin-6-yl or 7-amino-thieno [2,3c] Pyridin-2-yl,

W is alkyl (C = 1-8), diarylmethyl (C = 6-10), arylalkyl (C = 5-12), heteroaryl (C = 4-10), heteroarylalkyl (C = 5--10 11) dicycloalkylmethyl (C = 3-6) or cycloalkyl (C = 3-8),

X is hydrogen, alkyl (C = 1-8), cycloalkyl (C = 3-8), alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkylaminocarbonylalkyl, Monoalkylaminocarbonylalkyl, aminocarbonylalkyl, methanesulfonyl, aminosulfonyl, and O-alkyloxalyl,

Y is hydrogen or methyl,

n is 0, 1 or 2.

Representative compounds of the invention are as follows:

2- {2-[(1-Amino-isoquinolin-7-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid ethyl ester ;

2- {2-[(1-Amino-isoquinolin-7-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid ethyl ester ;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- Aminosulfonyl) amines;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- Methanesulfonyl) amine;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- [O-methyloxalyl]) amine;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamine;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- [Ethoxycarbonylmethanesulfonyl]) amine;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ¹ , N ¹ -dimethyl) amine;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- Cyanomethyl) amine;

2- {2-[(1-Amino-2-oxo-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl Amino acetic acid ethyl esters;

2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylaminosulfonyl acetic acid ;

2- {2-[(7-aminothieno [2,3c] pyridin-2-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) Ethylamino acetic acid ethyl ester.

The abbreviations for the reagents used in the examples and the like used in the following schemes are as follows:

PG: protection group

Boc: t-butoxycarbonyl

Alloc: allyloxycarbonyl

Me: methyl

Ph: phenyl

DMF: Dimethylformamide

THF: Tetrahydrofuran

TEA: Triethylamine

DIPEA: Diisopropylethylamine

EDC: 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride

HOBt: hydroxybenzotriazole

MeOH: Methanol

EA: ethyl acetate

AN: acetonitrile

LDA: lithium diisopropylamide

Compounds synthesized in the present invention are shown in the following reaction schemes divided into several representative categories.

Scheme 1 illustrates the synthesis of a compound represented by Formula 1 in a reverse synthetic manner. The compound of Formula 1 may be synthesized through one step or 2-3 step unit reaction in the compound of Formula 2. The representation of a primed (eg W ', X', Y ') group shown here is a generic representation of a precursor functional group that can be converted to an unprimed (eg W, X, Y) group. For example, an amine group to which a Boc group is attached can be seen as a precursor group of the amine group. However, not only the protecting group is referred to as a precursor group, but represents all groups obtained through alkylation, acylation, sulfonation and the like. Compound (2) can be obtained by reacting with compound (3) and amine (5). The compound in which R is hydrogen in compound (3) is obtained by hydrolyzing compound (6). These reactions may include reactions of alkylation, acylation, sulfonation and the like, including interconversion of protecting groups. Compound (6) is obtained by amidation of compound (7) and compound (8).

The amines (5) shown in Scheme 1 are the compounds (5a, 5b, 5c) shown below. They may be used as shown below, or in some cases, modified into prodrug forms. Specific forms thereof are described in detail in the experimental section.

Of these compounds (5a, 5b, 5c), compounds (5a, 5b) were synthesized by the method described in the paper or patent, and then used by protecting an arylamino group.

Synthesis of the compound (5c) was directly studied and synthesized in the course of this study, the specific method of synthesis is shown below. That is, thiophene-3-aldehyde was reacted with malonic acid to obtain compound (10), and the intermediate (11) was synthesized through the azide reaction. Intermediate (11) was heated to low temperature (˜80 ° C.) in diphenyl ether to obtain isocyanate (not shown), and heated again at high temperature (˜260 ° C.) to obtain compound (12). Compound (12) was treated with triphenylphosphine-bromine complex to obtain bromo compound (13), which was then reacted with LDA and DMF to give aldehyde (14). Compound (14) was reduced with NaBH 4 to obtain an alcohol compound (15), which was obtained by reacting phthalimide with Mitsunobu to obtain compound (16). Compound (16) was treated with hydrazine to deprotection to give compound (18), which was then protected to Boc group to give compound (19). Compound (19) was hydrolyzed to obtain carboxylic acid (20), which was subjected to Curtius rearrangement, and then allyl alcohol was reacted to obtain compound (21). Compound (21) was treated with trifluoroacetic acid to give compound (5c).

Known coupling reagents that can be used for the coupling reaction of amino groups include dicyclohexylcarbodiimide (DCC), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (EDC), bis- ( 2-oxo-3-oxazolidinyl) -phosphinic chloride (BOP-Cl), diphenylphosphoryl azide (DPPA), isobutylchloroformate, and the like, but are not limited to these.

As mentioned above, the compound of the formula (1) according to the present invention is a thrombin inhibitor which is better in selection for thrombin than the known compounds and is also likely by oral administration. Accordingly, the compounds of the present invention are useful for preventing blood clotting and for treating thrombosis.

Accordingly, the present invention provides a medicament for preventing blood coagulation and treating thrombosis containing a compound of Formula 1, a pharmaceutically acceptable salt thereof, a prodrug, a hydrate, a solvate or an isomer as an active ingredient together with a pharmaceutically acceptable carrier. It is another object to provide a pharmaceutical composition.

The total daily dose to be administered to the host in a single dose or in separate doses when administering a compound of the present invention for clinical purposes is preferably in the range of 0.001 mg to 10 mg per kg of body weight, but the specific dose level for a particular patient will be the specific compound to be used. May vary depending on body weight, sex, health status, diet, time of administration, method of administration, excretion rate, drug mixing and severity of disease.

The compounds of the present invention can be administered in injectable and oral formulations as desired.

Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, can be prepared using suitable dispersing agents, wetting agents, or suspending agents according to known techniques. Acceptable solvents that can be used include water, Ringer's solution and isotonic NaCl solution, and sterile fixed oils are conventionally employed as a solvent or suspending medium. Any non-irritating fixed oil may be used for this purpose, including mono- and diglycerides, and fatty acids such as oleic acid are used in the preparation of injectables.

Solid dosage forms for oral administration may be capsules, tablets, pills, powders and granules, and capsules and tablets are particularly useful. Tablets and pills are preferably prepared with the enteric agent. In the solid dosage form, the active compound of the formula (1) according to the present invention is mixed with one or more inert diluents such as sucrose, lactose, starch, etc., and a carrier such as a lubricant such as magnesium stearate, a disintegrant, a binder, and the like. It can manufacture.

One of the great features of the compound of Formula 1 according to the present invention is that the pharmaceutical composition containing the same has an effect even when formulated orally as an oral preparation. The results of kinetic experiments demonstrated that oral administration of the pharmaceutical compositions of the present invention confirmed the concentration of drug in the blood for a long time. Therefore, it is more useful in that it can be effectively used as an oral preparation, unlike conventional thrombin inhibitors.

On the other hand, when clinically administering the compound of the present invention to obtain the desired anticoagulant effect and thrombolytic effect, the active compound of formula 1 according to the present invention is one or more components selected from thrombolytic agents and platelet activity inhibitors It can be administered at the same time. Thrombolytic agents that can be administered in combination with a compound of the present invention in this manner may include T-PA, urokinase, Streptokinase, and the like, and platelet inhibitors include aspirin and ticlopidine. (Ticlopidin), Clopidrogel (Clopidrogel), 7E3 monoantibodies and the like.

However, the preparations containing the compounds according to the invention for the purpose of the treatment and prevention of thrombi are not limited to those described above, and any preparations useful for the treatment and prevention of thrombi can be included.

The present invention is explained in more detail by the following Preparation Examples, Examples and Experimental Examples, but the scope of the present invention is not limited in any way by these.

Preparation Example 1. Boc-D- (diphenylmethyl) alanine-proline-OH: compound [3a]

Boc-D- (diphenylmethyl) alanine-OH (3.01 g, 8.82 mmol), proline methylester hydrochloride (1.75 g, 1.2 equiv), EDC (2.54 g, 1.5 equiv), HOBt (1.79 g, 1.5 equiv) The solution was dissolved in about 40 mL of dimethylformamide, maintained at 0 ° C., and TEA (4.92 mL, 4.0 eq) was added thereto, followed by stirring at room temperature for one day. Concentrated at low pressure, the residue was taken up in ethyl acetate and then extracted-dried-condensed by conventional methods. Separation-purification using column chromatography (30% ethyl acetate / hexanes) gave 3.88 g (yield: 97%) of compound [6a] as a pale yellow oil.

The compound obtained above was dissolved in 50 mL of a mixed solvent (THF: MeOH: H ₂ O = 3: 2: 1), lithium hydroxide (0.51 g, 1.4 equivalents) was added thereto, and the mixture was stirred at room temperature for 3 hours. After neutralization with 6N HCl (2.2 mL), the solution was concentrated at low pressure, and the residue was dissolved in ethyl acetate and extracted, dried and concentrated by conventional methods. 3.76 g of the title compound [3a] was obtained quantitatively as a white powder.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 7.30-7.15 (m, 10H), 5.12 (m, 2H), 4.36 (m, 1H), 4.14 (m, 1H), 3.75 (m, 1H), 2.70 (m, 1H), 2.22 (m, 1H), 1.81 (m, 1H), 1.53 (m, 1H), 1.33 (s, 9H).

Preparation Example 2 Synthesis of Alloc-D- (Diphenylmethyl) alanine-Proline-OH (Compound [3b])

Boc-D- (diphenylmethyl) alanine-proline-OMe (3a) (1.43 g, 3.16 mmol) was dissolved in 3 mol of ethyl acetate and stirred at room temperature for 2 hours 30 minutes. Concentration under reduced pressure gave 1.23 g of D- (diphenylmethyl) alanine-proline-OMe quantitatively.

The compound obtained above was dissolved in 10 mL of dichloromethane, Na ₂ CO ₃ (610 mg, 2.3 equiv) and water (3 mL) were added, followed by cooling to 0 ° C. Allyl chloroformate (0.37 ml, 1.1 equiv) was added to the solution and stirred for 2 hours and 40 minutes at room temperature. To this solution was added 50 ml of dichloromethane, washed with 1N HCl (30 ml), saturated NaHCO ₃ (30 ml) and dried-concentrated in a conventional manner to quantitatively measure 1.38 g of Alloc-D- (diphenylmethyl) alanine-proline-OMe. Obtained.

The compound obtained above was dissolved in 20 mL of a mixed solvent (THF: MeOH: H ₂ O = 3: 2: 1), lithium hydroxide (0.28 g, 2.2 equiv) was added thereto, and the mixture was stirred at room temperature for one day. Neutralize with 1N HCl (6.5 mL) and concentrate at low pressure. The residue is dissolved in ethyl acetate and extracted, dried and concentrated by conventional method to give the title compound Alloc-D- (diphenylmethyl) alanine-proline-OH ( 3b) 1.31 g was obtained quantitatively.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 7.38-7.15 (m, 10H), 5.82-5.74 (m, 1H), 5.23 (d, 2H), 5.15-5.12 (m, 2H), 4.51-4.29 ( m, 2H), 4.38-4.36 (m, 1H), 4.12-4.06 (m, 1H), 3.77-3.75 (m, 1H), 3.68 (s, 3H), 2.82-2.78 (m, 1H), 1.81- 1.78 (m, 2H), 1.70 (m, 1H), 1.45 (m, 1H)

Preparation Examples 3 and 4. Synthesis of Compound [5a], [5b]

According to papers and patents (JBM Rewinkle et al./ Bioorganic & Medicinal Chemistry Letters 9 (1999) 685-690; Hendrickson, JB; Rodriguez, C .; J. Org. Chem. 1983,48 (19), 3344.) Synthesis was carried out, but the synthesis method was modified little by little if necessary.

Compound [5a] (Preparation Example 3):

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.45 (d, 1H), 8.12 (s, 1H), 7.93 (d, 1H), 7.90 (d, 1H), 7.83 (d, 1H), 4.27 (s , 2H), 1.30 (s, 18H)

Compound [5b] (Preparation Example 4):

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.43 (d, 1H), 8.25 (s, 1H), 8.01 (d, 1H), 7.88 (d, 1H), 7.64 (d, 1H), 4.46 (s , 2H), 1.30 (s, 18H)

Preparation Example 5 Synthesis of (2-Aminomethyl-thieno [2,3-c] pyridin-7-yl) -carbamic acid allyl ester (Compound [5c])

5-1) Synthesis of 3- [thiophen-3-yl] acrylic acid (Compound [10])

Thiophene-3-aldehyde [9] (50 g, 0.446 mol) and malonic acid (92.8 g, 2 equiv) were dissolved in pyridine (200 mL) well, then piperidine (4.5 mL) was added to the mixture at about 95-100 ° C. Heated for 22 hours. The temperature was raised to 145 ° C. and heated for 2 hours, then cooled. Most of the pyridine was removed by distillation under reduced pressure, and the yellow residue was added to ˜2N-HCl (150 mL), and extracted twice with ethyl acetate (300 mL, 200 mL). The extract was washed twice with dilute hydrochloric acid (100 mL × 2), dehydrated (anhydrous sodium sulfate), filtered (silica + celite pad) -decompressed. Trituration of the obtained solid with hexane gave 64.1 g (93.2%) of the title compound.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 7.77 (d, 1H), 7.55 (s, 1H), 7.35 (d, 1H), 7.32 (d, 1H), 6.27 (d, 1H)

5-2) Synthesis of 3- [thiophen-3-yl] acryloyl azide (Compound [11])

3- [thiophen-3-yl] acrylic acid [10] (64.0 g, 0.415 mole) and triethylamine (69.4 mL, 1.2 equiv) are dissolved in acetone (1 L) and cooled to below 0 ° C. in an ice-sodium chloride bath. Fit. To this was added dropwise an ethylchloroformate (51.6 mL, 1.3 equiv) -acetone (300 mL) solution (90 minutes required). After further stirring for 10 minutes, sodium azide (40.5 g, 1.5 equiv) -water (130 mL) solution was added dropwise (40 min required). After stirring for an additional 1.5 hours, it was poured into 6 L of ice water. The resulting solid was filtered, washed with water (2 L) and then dried through nitrogen gas for 3 days to give 64.9 g (87.3%) of the title compound.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 7.79 (d, 1H), 7.56 (s, 1H), 7.38 (d, 1H), 7.36 (d, 1H), 6.32 (d, 1H)

5-3) Synthesis of Thieno [2,3-c] pyridin-7-ol (Compound [12])

3- [thiophen-3-yl] acrylic acid azide [11] (32.4 g, 0.181 mole) was dissolved in 1.5 L of diphenyl ether and heated slowly to 105 ° C. for 3 hours. Tri-n-butyl amine (25.9 mL, 0.6 equiv) was added thereto and heated at 258 ° C. (reflux temperature) for 19 hours. After cooling, the resulting solid was filtered, washed with hexane and ether and dried to give 19.1 g (69.4%) of the title compound.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 12.35 (br s, 1H), 7.75 (d, 1H), 7.32 (d, 1H), 7.27 (d, 1H), 6.75 (d, 1H)

5-4) Synthesis of 7-bromo-thieno [2,3-c] pyridine (Compound [13])

Triphenylphosphine (33 g, 0.12 mol) was dissolved in 100 mL of dichloromethane and cooled with an ice water bath. Bromine (5.9 mL, 0.15 mol) was slowly added dropwise thereto. After dropping, the solvent was removed under reduced pressure, and monochloro benzene (60 mL) was added to dissolve it, and hydroxy pyridinothiophene (14.5 g 0.096 mol) was added thereto, and the reaction solution was heated to 140 ° C. and stirred for 3 hours. The reaction solution was cooled to room temperature and poured into ice water (200 g), followed by adding 2 normal caustic soda solution to adjust the pH to 10, and extracting with ethyl acetate (300 mL). Purification using a silica gel column followed by drying gave the title compound (16 g, yield 60%).

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.27 (d, 1H), 7.76 (d, 1H), 7.67 (d, 1H), 7.47 (d, 1H)

5-5) Synthesis of (7-bromo-thieno [2,3-c] pyridin) -2-yl-methanol (Compound [15])

Diisotropilamine (9.5 mL) was dissolved in 100 mL of tetrahydrofuran and cooled to -30 ° C in a dry ice bath. N-butylamine (25 mL, 2.5 M nucleic acid solution) was added dropwise thereto and stirred for 30 minutes. A solution of 7-bromo pyridinothiophene [13] (11.6 g, 0.05 mol) obtained in Scheme 5-4) was added to 20 mL of tetrahydrofuran, cooled to -78 ° C, and stirred for 30 minutes. Formaldehyde (6.0 mL) was added dropwise and stirred for 30 minutes. The reaction solution was slowly poured into 1 normal hydrochloric acid solution cooled with ice, extracted with ethyl acetate (300 mL), and concentrated to dryness in a conventional manner. The solid obtained here was dissolved again in a mixed solution of methanol (100 mL) and tetrahydrofuran (100 mL), cooled to −20 ° C., and sodium borohydride (2.4 g) was added thereto, followed by stirring for 1 hour. The reaction solution was slowly poured into 1 normal hydrochloric acid cooled with ice, extracted with ethyl acetate (300 mL), and concentrated to dryness in the usual manner to obtain the title compound (12.8 g) in a yield of 90%.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.24 (d, 1H), 7.56 (d, 1H), 7.33 (s, 1H), 5.00 (s, 2H)

5-6) Synthesis of 2- (7-bromo-thieno [2,3-c] pyridin-2-ylmethyl) -isoindole-1,3-dione (Compound [16])

Triphenylphosphine (16.5 g), phthalimide (8.48 g) and the title compound (7-bromo-thieno [2,3-c] pyridin) -2-yl-methanol) The mixture was dissolved in dry tetrahydrofuran (300 mL), cooled to 0 ° C. with an ice bath, diethyl azodicarboxylate (10.8 mL) was added dropwise, the ice bath was removed, and the mixture was stirred at room temperature for 5 hours. The reaction solution was concentrated under reduced pressure and separated and purified through a silica gel column to obtain the title compound (11.2 g) in 60% yield.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.23 (d, 1H), 7.88 (dd, 2H), 7.74 (dd, 2H), 7.56 (d, 1H), 7.47 (s, 1H), 5.12 (s , 2H)

5-7) 2- (1,3-dioxo-1,3-dihydro-isoindol-2-ylmethyl) -thieno [2,3-c] pyridine-7-carboxylic acid ethyl ester (compound [17]) Synthesis

The title compound (11.6g) of Preparation Example 5-6) was dissolved in a mixed solvent of ethanol (50 mL) and dimethyl sulfoxide (50 mL), and then 1,3-bisdiphenylphosphinopropane (3.2 g) and palladium acetate (1.4 g) were dissolved. ) And triethylamine (90 mL) were added successively, and the mixture was heated to 80 ° C and stirred for 5 hours while supplying a carbon monoxide gas filled with a hot air balloon. After completion of the reaction, 400 mL of water was added, followed by extraction with ethyl acetate (500 mL) and separation and purification through a silica gel column to obtain the title compound (6.7 g, 70% yield).

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.64 (d, 1H), 7.87 (dd, 2H), 7.77 (dd, 2H), 7.74 (d, 1H), 7.42 (s, 1H), 5.16 (s , 2H), 4.56 (q, 2H), 1.47 (t, 3H)

5-8) Synthesis of 2-aminomethyl-thieno [2,3-c] pyridine-7-carboxylic acid ethyl ester (Compound [18])

The title compound (1.7 g) of Preparation Example 5-7 was dissolved in ethanol (100 mL), and then hydrazine monohydrate (1.8 mL) was added thereto, followed by stirring for 6 hours while heating to reflux. The reaction solution was cooled to room temperature, and the resulting solid was removed by filtration. The filtrate was concentrated and separated and purified through a silica gel column to obtain the title compound (0.98 g, 86% yield).

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.38 (d, 1H), 7.67 (d, 1H), 7.32 (s, 1H), 4.72 (Abq, 2H), 4.43 (q, 2H), 1.18 (t, 3H)

5-9) Synthesis of 2- (t-butoxycarbonylamino-methyl) -thieno [2,3-c] pyridine-7-carboxylic acid ethyl ester (Compound [19])

The title compound (3.5 g, 14.8 mM) of Preparation Example 5-8) was dissolved in a mixed solvent of dioxane (80 mL) and water (20 mL), followed by di-t-butyldicarbonate (4.85 g) and sodium carbonate (2.35 g). ) Was added sequentially, followed by stirring at room temperature for 4 hours. Water (300 mL) was added thereto, extracted with ethyl acetate (300 mL), and the organic solvent layer obtained by washing with saturated brine was concentrated under reduced pressure, and then purified by passing through a silica gel column to obtain the title compound (3.6 g, 68% yield). Obtained.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.45 (d, 1H), 7.57 (d, 1H), 7.19 (s, 1H), 5.12 (bs, 1H), 4.59 (Abq, 2H), 4.56 (q , 2H), 1.48 (t, 3H), 1.45 (s, 9H)

5-10) Synthesis of 2- (t-butoxycarbonylamino-methyl) -thieno [2,3-c] pyridine-7-carboxylic acid (compound [20])

The title compound (3.5g, 10.4mM) of Preparation Example 5-9) was dissolved in a mixed solvent of tetrahydrofuran (40 mL) and water (10 mL), and lithium hydroxide monohydrate (656 mg) was added thereto, followed by stirring at room temperature for 1 hour. It was. To this was added 1 normal aqueous hydrochloric acid solution (200 mL), extracted with ethyl acetate (300 mL), and concentrated to dryness in the usual manner to obtain the title compound (3.2 g, 97% yield).

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.57 (d, 1H), 7.85 (d, 1H), 7.30 (s, 1H), 5.98 (bs, 1H), 4.57 (Abq, 2H), 1.37 (s , 9H)

5-11) Synthesis of 2- (t-butoxycarbonylamino-methyl) -thieno [2,3-c] pyridine-7-allyloxycarbonylamine (Compound [21])

1.3 g of the title compound obtained in Preparation Example 5-10) was dissolved in 10 ml of benzene, 0.7 ml of triethylamine was added, followed by stirring for 10 minutes. 10 ml of tetrahydrofuran and 1.0 ml of diphenylphosphoryl azide were added thereto, and the reaction temperature was raised to 40 ° C., stirred for 1 hour, and heated to reflux for 3 hours. The reaction solution was concentrated and purified through a silica gel column to give 1.2 g of the title compound in 91% yield.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.59 (d, 1H), 7.85 (d, 1H), 7.30 (s, 1H), 5.98 (bs, 1H), 5.31 (m, 1H), 4.96 (dd) , 1H), 4.94 (dd, 1H), 4.65 (d, 2H), 4.57 (Abq, 2H), 1.36 (s, 9H)

5-12) Synthesis of 2-aminomethyl-thieno [2,3-c] pyridine-7-allyloxycarbonylamine (Compound [5c])

1.2 g of the title compound obtained in Preparation Example 5-11) was dissolved in 10 ml of dichloromethane, and 5 ml of trifluoroacetic acid was added thereto, followed by stirring at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, 100 ml of hexane was slowly added, and the resulting solid was filtered and dried to yield 800 mg of the title compound in a yield of 95%.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.57 (d, 1H), 7.88 (d, 1H), 7.33 (s, 1H), 5.94 (bs, 1H), 5.28 (m, 1H), 4.996 (dd , 1H), 4.97 (dd, 1H), 4.66 (d, 2H), 4.53 (Abq, 2H)

Preparation Example 6. [2- (2-{[1- (N, N'-di-t-butoxycarbonyl-amino) -isoquinolin-6-ylmethyl] -carbamoyl} -pyrrolidine-1 -Yl) -2-oxo- (1-diphenylmethyl) ethyl] allyloxycarbonylamine (Compound 2-1)

Alloc-D- (diphenylmethyl) alanine-proline-OH (4 g) and compound (5a) (3.9 g) were dissolved in dichloromethane (30 mL) and cooled to 0 ° C. with an ice water bath, then 1- (3- Dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (2.2 g), hydroxybenzotriazole (1.5 g) and triethylamine (3.3 mL) were added sequentially and the ice water bath was removed to raise the reaction temperature to room temperature and 3 Stirred for time. To this was added 1 normal hydrochloric acid solution (200 mL), extracted with ethyl acetate (200 mL), washed with saturated aqueous sodium bicarbonate solution (200 mL), dried and concentrated by a conventional method, and purified by passing through a silica gel column. The yield of the title compound (6.1 g) was obtained.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.37 (d, 1H), 7.87 (d, 1H), 7.71 (s, 1H), 7.67 (t, 1H), 7.62 (d, 1H), 7.47 (m , 1H), 7.40-7.20 (m, 10H), 5.51-5.43 (m, 1H), 5.22-5.12 (m, 1H), 4.95-4.87 (m, 3H), 4.75 (dd, 1H), 4.46-4.39 (m, 3H), 4.05 (dd, 1H), 3.84 (dd, 1H), 3.74-3.66 (m, 1H), 2.54-2.51 (m, 1H), 2.15 (m, 1H), 1.73 (m, 1H ), 1.45 (m, 1H), 1.29 (s, 18H)

Preparation Example 7 [2- (2-{[1- (N, N'-di-t-butoxycarbonyl-amino) -isoquinolin-7-ylmethyl] -carbamoyl} -pyrrolidine-1 -Yl) -2-oxo- (1-diphenylmethyl) ethyl] allyloxycarbonylamine (Compound 2-2)

3.9 g of compound [5b] was used instead of compound [5a] used in Preparation Example 6 to obtain 5.9 g of the title compound in a yield of 79%.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.37 (d, 1H), 7.81 (d, 1H), 7.74 (s, 1H), 7.62 (m, 2H), 7.58 (d, 1H), 7.40-7.19 (m, 10H), 5.54-5.48 (m, 1H), 5.11 (d, 1H), 4.96-4.93 (m, 2H), 4.87 (dd, 1H), 4.64 (dd, 1H), 4.53 (dd, 1H ), 4.42 (d, 1H), 4.39 (d, 1H), 4.09 (dd, 1H), 3.83 (dd, 1H), 3.74-3.66 (m, 1H), 2.53 (m, 1H), 2.08 (m, 1H), 1.77 (m, 1H), 1.45 (m, 1H), 1.30 (s, 18H)

Preparation Example 8 [2- (2-{[7- (allyloxycarbonylamino) -thieno [2,3-c] -2-ylmethyl] -carbamoyl} -pyrrolidin-1-yl) 2-oxo- (1-diphenylmethyl) ethyl] -t-butoxycarbonylamine (Compound 2-3)

3.7 g of compound [5c] was used instead of compound [5a] used in Preparation Example 6 to obtain the title compound in a yield of 78%.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.57 (d, 1H), 7.88 (d, 1H), 7.40-7.19 (m, 10H), 7.33 (s, 1H), 5.94 (bs, 1H), 5.28 (m, 1H), 4.996 (dd, 1H), 4.97 (dd, 1H), 4.87 (dd, 1H), 4.66 (d, 2H), 4.53 (Abq, 2H) 4.87 (dd, 1H), 4.64 (dd , 1H), 4.53 (dd, 1H), 4.42 (d, 1H), 4.39 (d, 1H), 4.09 (dd, 1H), 3.83 (dd, 1H), 3.74-3.66 (m, 1H), 2.53 ( m, 1H), 2.08 (m, 1H), 1.77 (m, 1H), 1.45 (m, 1H), 1.30 (s, 18H)

Preparation Example 9. 2- (2-{[1- (N, N'-di-t-butoxycarbonyl-amino) -isoquinolin-6-ylmethyl] -carbamoyl} -pyrrolidine-1- Synthesis of yl) -2-oxo- (1-diphenylmethyl) ethylamino acetic acid ethyl ester (Compound 2-4)

[2- (2-{[1- (N, N'-di-t-butoxycarbonyl-amino) -isoquinolin-6-ylmethyl] -carbamoyl} -pyrrolidin-1-yl)- 2-oxo- (1-diphenylmethyl) ethyl] allyloxycarbonylamine (3.95 g) was dissolved in a mixed solvent of dichloromethane (50 mL) and dimethylformaldehyde (50 mL), followed by phenylsilane (0.8 mL) and dichlorobis. Triphenylphosphinepalladium (0.18 g) was added sequentially and stirred at room temperature for 5 hours. The reaction solution was concentrated under reduced pressure, 350 mg of the solid obtained by separation and purification through a silica gel column was collected, and then dissolved in dichloromethane (20 ml). Diisopropylethylamine (0.13 mL) and ethyl bromoacetate (0.65 mL) were sequentially added thereto, followed by stirring for 10 hours. The reaction solution was concentrated under reduced pressure and separated and purified by silica gel column to give the title compound (280 mg) in 70% yield.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.36 (d, 1H), 7.87 (d, 1H), 7.75 (s, 1H), 7.64 (m, 2H), 7.55 (d, 1H), 7.42-7.17 (m, 10H), 4.83 (dd, 1H), 4.64 (dd, 1H), 4.53 (dd, 1H), 4.42 (d, 1H), 4.39 (d, 1H), 4.09 (dd, 1H), 3.91 ( q, 2H), 3.83 (dd, 1H), 3.74-3.66 (m, 1H), 2.74 (Abq, 2H), 2.53 (m, 1H), 2.08 (m, 1H), 1.77 (m, 1H), 1.45 (m, 1H), 1.23 (t, 3H), 1.30 (s, 18H)

Preparation Example 10 1- (2-Methanesulfonylamino-3,3-diphenyl-propionyl) -pyrrolidine-2-carboxylic acid (1-N, N'-di-t-butoxycarbonyl- Synthesis of Amino-isoquinolin-6-ylmethyl) -amide (Compound 2-5)

The title compound (290 mg) was obtained using methanesulfonyl chloride (0.21 mL) instead of the ethyl bromoacetate used in Preparation Example 9 in the same manner as in Preparation Example 9.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 8.36 (d, 1H), 7.85 (d, 1H), 7.73 (s, 1H), 7.64 (m, 2H), 7.55 (d, 1H), 7.42-7.18 (m, 10H), 4.83 (dd, 1H), 4.64 (dd, 1H), 4.53 (dd, 1H), 4.42 (d, 1H), 4.39 (d, 1H), 4.09 (dd, 1H), 3.83 ( dd, 1H), 3.74-3.66 (m, 1H), 2.83 (s, 3H), 2.54 (m, 1H), 2.09 (m, 1H), 1.76 (m, 1H), 1.45 (m, 1H), 1.30 (s, 18 H)

Preparation Example 11 (2- {2-[(1-N, N'-di-t-butoxycarbonyl-amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl } -1-synthesis of benzhydryl-2-oxo-ethylamino) -acetic acid (compound 2-6)

The title compound (140 mg) of Preparation Example 9 was dissolved in a mixed solvent of tetrahydrofuran (4 mL) and water (1 mL), and lithium hydroxide monohydrate (20 mg) was added thereto, followed by stirring at room temperature for 1 hour. To this was slowly added 4 normal hydrochloric acid ethanol solution to adjust the pH to 3, distilled under reduced pressure and dried to give the title compound (yield: quantitative) which was used for the next reaction without further purification.

¹ H-NMR (CDCl ₃ , 400 MHz) δ 8.38 (d, 1H), 7.89 (d, 1H), 7.75 (s, 1H), 7.64 (m, 2H), 7.55 (d, 1H), 7.42-7.17 (m, 10H), 4.85 (dd, 1H), 4.66 (dd, 1H), 4.52 (dd, 1H), 4.42 (d, 1H), 4.39 (d, 1H), 4.09 (dd, 1H), 3.83 ( dd, 1H), 3.74-3.66 (m, 1H), 2.74 (Abq, 2H), 2.53 (m, 1H), 2.08 (m, 1H), 1.76 (m, 1H), 1.44 (m, 1H), 1.32 (s, 18 H)

Example 1. (2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl Amino acetic acid ethyl ester: synthesis of 2 trifluoroacetic acid salt (compound 1-1)

The title compound (105 mg) of Preparation Example 9 was dissolved in a mixed solution of trifluoroacetic acid (3 mL) and dichloromethane (7 mL), and stirred at room temperature for 30 minutes. The concentrated solution obtained by concentrating the reaction solution under reduced pressure was separated and purified by HPLC, and the aliquot obtained was lyophilized to obtain the title compound (80 mg) in a yield of 75%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.96 (br s, 2H), 8.61 (d, 1H), 8.45 (d, 1H), 7.79 (s, 1H), 7.62 (d, 1H), 7.49-7.18 (m, 10H), 7.14 (d, 1H), 4.81 (d, 1H), 4.46 (ABq, 2H), 4.32 (d, 1H), 4.01 (q, 2H), 3.98 (m, 1H) , 3.53 (m, 1H), 3.01 (m, 1H), 2.45 (ABq, 2H), 1.71 (m, 3H), 1.31 (m, 1H), 1.07 (t, 3H)

Example 2.2- {2-[(1-Amino-isoquinolin-7-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino ethyl acetate Esters: Trifluoroacetic Acid (Compound 1-2)

106 mg (61% yield) of the compound obtained in 173 mg of the title compound of Preparation Example 7 was obtained in the same manner as in Example 9, to obtain 67 mg of the title compound in the yield of 94%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.71 (t, 1H), 8.43 (s, 1H), 7.91 (d, 1H), 7.83 (d, 1H), 7.66 (d, 1H), 7 , 57 (d, 2H), 7.40-7.20 (m, 10H), 4.78 (m, 1H), 4.56 (dd, 1H), 4.32 (m, 2H), 4.01-3.90 (m, 3H), 3.60-3.45 (m, 3H), 3.08 (m, 1H), 1.85-1.65 (m, 3H), 1.30 (m, 1H), 1.01 (t, 3H)

Example 3.2- {2-[(1-Amino-isoquinolin-7-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid: 2 trifluoroacetic acid salt (compound 1-3)

185 mg (64% yield) of the compound obtained in 299 mg of the title compound of Preparation Example 7 in the same manner as in Preparation Example 9 were sequentially subjected to the methods of Preparation Example 11 and Example 1 to obtain 100 mg of the title compound in a yield of 87%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.72 (t, 1H), 8.45 (s, 1H), 7.90 (d, 1H), 7.82 (d, 1H), 7.66 (d, 1H), 7 , 60 (d, 2H), 7.40-7.20 (m, 10H), 4.93 (m, 1H), 4.56 (dd, 1H), 4.37 (m, 2H), 3.98 (m, 1H), 3.60-3.45 (m , 3H), 3.11 (m, 1H), 1.82-1.65 (m, 3H), 1.31 (m, 1H)

Example 4.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid: 2 trifluoroacetic acid salt (compound 1-4)

85 mg of the compound synthesized in Preparation Example 11 was used in place of the title compound of Preparation Example 9 used in Example 1 in a similar manner to Example 1 to give 72 mg (80% yield) of the title compound.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 9.00 (br s, 2H), 8.65 (d, 1H), 8.46 (d, 1H), 7.77 (s, 1H), 7.61 (d, 1H), 7.52-7.19 (m, 10H), 7.17 (d, 1H), 4.95 (d, 1H), 4.45 (ABq, 2H), 4.39 (d, 1H), 3.99 (m, 1H), 3.49 (m, 1H) , 3.00 (m, 1H), 2.45 (ABq, 2H), 1.72 (m, 3H), 1.29 (m, 1H)

Example 5.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ¹ -Aminosulfonyl) amine: 2 trifluoroacetic acid salt (compound 1-5)

From 150 mg of the compound obtained in Preparation Example 6, 140 mg (82% yield) of the compound obtained using a method similar to Preparation Example 9 (using chloro sulfonamide (0.3 ml) instead of ethyl bromo acetate) was prepared in a similar manner to Example 1. Used in place of the title compound of Preparation Example 9 used in Example 1 to give 120 mg of the title compound in a yield of 84%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 9.03 (br s, 2H), 8.54 (d, 1H), 8.29 (d, 1H), 7.81 (s, 1H), 7.72 (d, 1H), 7.34-7.18 (m, 10H), 6.56 (d, 1H), 4.81 (d, 1H), 4.40 (ABq, 2H), 4.32 (d, 1H), 3.93 (m, 1H), 3.72 (m, 1H) , 2.94 (m, 1H), 1.72 (m, 2H), 1.54 (m, 1H), 1.36 (m, 1H)

Example 6.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ¹ Methanesulfonyl) amine: 2 trifluoroacetic acid salt (compound 1-6)

81 mg of the compound synthesized in Preparation Example 10 was used in place of the title compound of Preparation 9 used in Example 1 in a similar manner to Example 1 to yield 76 mg (78% yield) of the title compound.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 7.72 (s, 1H), 7.58 (d, 1H), 7.42 (d, 1H), 7.38-7.22 (m, 10H), 6.99 (d, 1H), 6.81 (d, 1H), 4.85 (d, 1H), 4.43 (ABq, 2H), 4.33 (d, 1H), 4.17 (m, 1H), 3.78 (m, 1H), 2.86 (s, 3H), 2.80 ( m, 1H), 1.99 (m, 1H), 1.86 (m, 1H), 1.74 (m, 1H), 1.49 (m, 1H)

Example 7.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ¹ -[O-methyloxalyl]) amine: 2 trifluoroacetic acid salt (compound 1-7)

From 90 mg of the compound obtained in Preparation Example 6, 90 mg (84% yield) of the compound obtained using a method similar to Preparation Example 9 (using methyl oxalyl chloride (0.34 ml) instead of ethyl bromo acetate) was prepared in a similar manner to Example 1. Used in place of the title compound of Preparation 9 used in Example 1 to give 88 mg of the title compound in a yield of 83%.

¹ H-NMR (CDCl ₃ , 500 MHz) δ 7.88 (d, 1H), 7.56 (d, 1H), 7.54 (s, 1H), 7.40 (d, 1H), 7.38-7.23 (m, 10H), 6.91 (d, 1H), 5.07 (d, 1H), 4.53 (d, 1H), 4.51 (ABq, 2H), 4.33 (d, 1H), 3.80 (m, 1H), 3.58 (s, 3H), 2.63 ( m, 1H), 2.11 (m, 1H), 1.53 (m, 3H)

Example 8.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamine: 2 Trifluoroacetate (Compound 1-8)

372 mg of the title compound of Preparation Example 6 was dissolved in a mixed solvent of dichloromethane (5 mL) and dimethyl formaldehyde (5 mL), and then phenylsilane (0.07 mL) and dichlorobistriphenylphosphinepalladium (17 mg) were added sequentially, followed by 5 hours at room temperature. Stirred. The reaction solution was concentrated under reduced pressure, and purified through a silica gel column to obtain 270 mg (82%) of solid. 100 mg of this solid was subjected to the method of Example 1 to give 73 mg of the title compound in a yield of 70%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.94 (br s, 1H), 8.45 (d, 1H), 8.26 (br s, 2H), 7.75 (s, 1H), 7.65 (m, 3H) , 7.45-7.19 (m, 10H), 5.01 (d, 1H), 4.72 (d, 1H), 4.39 (d, 1H), 3.95 (m, 1H), 3.53 (m, 1H), 2.87 (m, 1H ), 1.72 (m, 2H), 1.50 (m, 1H), 1.23 (m, 1H)

Example 9.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ¹ -[Ethoxycarbonylmethanesulfonyl] amine: 2 trifluoroacetic acid salt (compound 1-9)

From 115 mg of the compound obtained in Preparation 6, 115 mg (47% yield) of Compound obtained using a method similar to Preparation 9 (using ethoxycarbonylmethanesulfonyl chloride (81 mg) instead of ethyl bromo acetate) was used. A similar method was carried out to give 45 mg of the title compound in 43% yield.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.99 (br s, 2H), 8.59 (t, 1H), 8.46 (d, 1H), 8.13 (d, 1H), 7.84 (d, 1H), 7.63 (m, 2H), 7.48 (d, 1H), 7.40-7.20 (m, 10H), 5.01 (dd, 1H), 4.51 (dd, 1H), 4.34 (m, 2H), 4.15 (d, 1H) , 3.94-3.89 (m, 3H), 3.68 (d, 1H), 3.59 (m, 1H), 3.07 (m, 1H), 1.80 (m, 1H), 1.70-1.58 (m, 2H), 1.31 (m , 1H), 0.99 (t, 3H)

Example 10.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ¹ , N ¹ -dimethyl) amine: 2 trifluoroacetic acid salt (compound 1-10)

45 mg (44% yield) of a compound obtained using 100 mg of the compound obtained in Preparation Example 6 using a method similar to Preparation Example 9 using dimethyl sulfate (2 equivalents instead of ethyl bromo acetate and dimethylformamide instead of dichloromethane) was obtained. A method similar to Example 1 was carried out to give 34 mg of the title compound in a yield of 71%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 9.09 (br s, 2H), 8.76 (t, 1H), 8.48 (d, 1H), 7.80 (s, 1H), 7.77 (d, 1H), 7.67-7.64 (m, 2H), 7.45-7.16 (m, 10H), 5.53 (br s, 1H), 4.68 (d, 1H), 4.52 (dd, 1H), 4.42 (dd, 1H), 3.93 (m , 1H), 3.68 (m, 1H), 3.25 (m, 1H), 2.77 (br s, 6H), 1.90-1.78 (m, 2H), 1.74-1.65 (m, 1H), 1.35 (m, 1H)

Example 11.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ¹ -Cyanomethyl) amine: 2 trifluoroacetic acid salt (compound 1-11)

From 65 mg of the compound obtained in Preparation Example 6, 65 mg (61% yield) of the compound obtained using a method similar to Preparation Example 9 (using bromoacetonitrile instead of ethyl bromo acetate) was carried out in a similar manner to Example 1 to obtain the title compound. 37 mg was obtained with a yield of 55%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.91 (br s, 2H), 8.53 (t, 1H), 8.47 (d, 1H), 7.82 (s, 1H), 7.64 (m, 2H), 7.43 (d, 2H), 7.35-7.16 (m, 9H), 4.45 (d, 1H), 4.38 (d, 1H), 4.16 (d, 1H), 4.02 (m, 1H), 3.63 (m, 1H) , 3.43 (m, 2H), 3.11 (m, 1H), 1.83-1.68 (m, 3H), 1.37 (m, 1H)

Example 12.2- {2-[(1-Amino-2-oxo-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) Ethylamino acetic acid ethyl ester: 2 trifluoroacetic acid salt (compound 1-12)

185 mg of the title compound of Preparation Example 9 was dissolved in 5 mL of dichloromethane, and 62 mg of metachloroperbenzoic acid was added thereto, followed by stirring at room temperature for 2 days. 20 mL of dichloromethane was added thereto, washed with saturated sodium sulfite solution (20 mL) and saturated aqueous sodium bicarbonate solution (20 mL), and 188 mg of solid obtained by dry-concentration according to the conventional method was obtained. Obtained in the yield.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 9.10 (br s, 2H), 8.65 (d, 1H), 8.47 (d, 1H), 8.01 (d, 1H), 7.80 (d, 2H), 7.68-7.18 (m, 11H), 4.81 (d, 1H), 4.46 (ABq, 2H), 4.32 (d, 1H), 4.01 (q, 2H), 3.98 (m, 1H), 3.53 (m, 1H) , 3.01 (m, 1H), 2.45 (ABq, 2H), 1.71 (m, 3H), 1.31 (m, 1H), 1.07 (t, 3H)

Example 13.2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylaminosulfonyl Acetic acid: 2 trifluoroacetic acid salt (Compound 1-13)

120 mg (57% yield) of the compound obtained using 120 mg of the compound obtained in Preparation Example 6 using a method similar to Preparation Example 9 (using ethoxycarbonylmethanesulfonyl chloride instead of ethyl bromo acetate) was prepared. And 70 mg of solid were obtained. This solid was subjected to the method of Example 1 to yield 25 mg of the title compound in a yield of 30%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.99 (br s, 1H), 8.57 (t, 1H), 8.46 (d, 1H), 8.06 (d, 1H), 7.83 (s, 1H), 7.62 (d, 2H), 7.46 (d, 2H), 7.36 (t, 2H), 7.30-7.20 (m, 5H), 5.01 (dd, 1H), 4.51 (dd, 1H), 4.34 (m, 2H) , 4.10 (d, 1H), 3.94-3.89 (m, 1H), 3.60 (d, 1H), 3.59 (m, 1H), 3.07 (m, 1H), 1.80 (m, 1H), 1.70-1.58 (m , 2H), 1.31 (m, 1H)

Example 14.2- {2-[(7-aminothieno [2,3-c] pyridin-2-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-di Phenylmethyl) ethylamino acetic acid ethyl ester: 2 trifluoroacetic acid salt (Compound 1-14)

620 mg of a compound obtained in a similar manner to Example 9 using 450 mg of the title compound of Preparation Example 8 instead of Alloc-D- (diphenylmethyl) alanine-proline-OH used in Preparation Example 9 was prepared in a similar manner to Example 1 Was removed to give 400 mg of the title compound in a yield of 87%.

¹ H-NMR (DMSO-d ₆ , 500 MHz) δ 8.64 (d, 1H), 8.41 (d, 1H), 7.79 (s, 1H), 7.62 (d, 1H), 7.49-7.18 (m, 10H) , 7.14 (d, 1H), 4.81 (d, 1H), 4.46 (ABq, 2H), 4.32 (d, 1H), 4.01 (q, 2H), 3.98 (m, 1H), 3.53 (m, 1H), 3.01 (m, 1H), 2.45 (ABq, 2H), 1.71 (m, 3H), 1.31 (m, 1H), 1.07 (t, 3H)

Experimental Example: Analysis of Biological Activity of Thrombin Inhibitor

The thrombin inhibitory effect of the compound of general formula (1) according to the present invention is dissociated using the following formula according to the method described in Methods (Enzymology V.80 p341-361; Biochemistry 27 p2144-2151,1988). Measure by determining the constant Ki value.

Ki = [E]. [I] / [EI]

[E]: concentration of enzyme not bound to inhibitor

[I]: concentration of inhibitor not bound to enzyme

[EI]: concentration of enzyme and inhibitor combination

Since the dissociation constant Ki indicates the degree of dissociation between the enzyme and the thrombin inhibitor compound, the smaller the dissociation constant value, the greater the binding of the inhibitor to the enzyme, and thus the higher the inhibitory activity. These dissociation constants can be obtained by reacting with a specific substrate that exhibits color development when hydrolyzed by the action of thrombin and measuring the color development as a function of time according to spectrophotometry.

In the present invention, a chromozyme TH (Chromozyme TH: Tosyl-Gly-Pro-Arg-4-nitroanilide acetate) is used as a substance that develops upon hydrolysis under the action of thrombin as a substrate of thrombin. Hydrolysis of chromozyme TH by thrombin produces yellow para-nitroanilide. Therefore, the thrombin inhibitory activity of the compound according to the present invention can be measured by measuring the amount of para-nitroaniline produced by the change in absorbance with time. That is, the activity of the enzyme can be measured from the change in absorbance, which can be directly related to the ability of the thrombin inhibitor to inhibit the enzyme activity.

On the other hand, in order to determine the selectivity for thrombin inhibition relative to trypsin of the compound according to the present invention was carried out in the same manner as the method for measuring the thrombin inhibitory activity was measured by Ki value of the inhibitory effect of the compound of formula (1) to trypsin Obtain the following trypsin / thrombin ratio. At this time, the test method for trypsin is the same as thrombin, except that N-benzoyl-valine-glycine-arginine para-nitroanilide hydrochloride (N-benzoyl-Val-Gly-Arg-p-nitroanilide hydrochloride) ).

The ability of the compounds according to the invention to inhibit thrombus formation was measured in animal models.

Animals used were Sprague Dawley, 250-300 grams of body weight, raised in standard laboratory animals at 20-22 ° C, 12-hour contrast in the laboratory of LG Chem Institute of Technology. -4 animals were used. First, rats were anesthetized by intraperitoneal injection of urethane at a dose of 1.3 grams per kilogram of body weight. The abdomen of the rat was dissected to expose the inferior vena cava, and then sutures were spread over the vein just below the left renal vein and 1.6 cm below. 5 minutes after the compound of the example was injected through the left femoral vein, thromboplastin was also injected through the femoral vein. Subsequently, the sutures laid down after 30 seconds were ligated. Fifteen minutes after the suture was ligated, the veins were removed and the thrombus generated was taken and weighed.

The absorption effect upon oral administration of the compound according to the present invention was determined by measuring blood drug concentration according to the following experimental method. The male rats and the dogs were fasted for 18 hours each, and then tested. A 1% (10 mg / ml) solution of the compound of Example was prepared using an appropriate dissolution aid and then administered orally. Blood was collected at predetermined time intervals, followed by liquid extraction with methylene chloride, back extraction with dilute hydrochloric acid solution, and blood drug concentrations were measured by high pressure liquid chromatography (HPLC).

Experimental Example 1. Inhibitory Activity of Thrombin Inhibitor

The inhibitory capacity for thrombin activity of the compounds according to the invention was measured as described below.

Add 140 μl of 0.1M Tris buffer (pH 7.8) containing 150 mM NaCl, 0.1% PEG 8000 (polyethylene glycol, molecular weight about 8,000) to the microplate. The substrate solution was prepared by dissolving chromozyme TH in dimethyl sulfoxide (DMSO) at a concentration of 10 mM and then diluting with the buffer solution to a concentration of 0.5 mM. 20 μl of the 0.5 mM substrate solution thus prepared is added to the microplate. As an inhibitor solution, the inhibitor compound according to the present invention was dissolved in 10 mM of dimethyl sulfoxide, and then diluted with the buffer solution. Take it up and add to the microplate to a total volume of 180 μl.

At room temperature, 20 µl of human thrombin dissolved in the Tris buffer solution at a concentration of 0.1 NIH Unit was added to the microplates containing the reaction solution to start the enzymatic hydrolysis reaction. The amount of para-nitroanilide produced by the reaction for 10 minutes from the moment of addition of the enzyme was monitored by the change in absorbance at 381 nm, showing the continuous spectrum of reaction time versus absorbance. The above experiments were performed for different inhibitor concentrations to obtain continuous spectra.

After calculating the initial velocity Vi from the slope within 30 seconds of the initial reaction time in each spectrum, an inverse graph of the initial velocity versus the inhibitor concentration (1 / Vi) is shown. After calculating the linear equation that satisfies the points on the graph, Ki can be calculated from the x-intercept of the equation using the enzyme reaction equation. The K _m value used in this calculation was obtained by varying the substrate concentration at a constant enzyme concentration of 10 μM.

On the other hand, the inhibitory activity of the compound according to the present invention against trypsin was also measured as described for the case of thrombin.

As a substrate, 20 µl of a 500 µM solution of N-benzoyl-Val-Gly-Arg-p-nitroanilide hydrochloride was used, and the inhibitor was used at 0. Various concentrations were used within the 100 μM range. In addition, trypsin was dissolved in 0.1 N HCl was made to 1 μg / ㎖ with the Tris buffer solution immediately before the experiment and 20 μl was used. As in the experiment for thrombin, the total volume of the reaction solution was set to 200 µl and in the same manner. The K _m value used for the calculation of Ki was determined by the same method, which was 160 µM.

The enzymatic activity of the inhibitor according to the present invention measured for thrombin and trypsin according to the method described above is expressed in Ki values and the selectivity to thrombin is expressed in trypsin / thrombin. The results are as shown in Table 1 below.

Experimental Example 2 Inhibition of Thrombosis in Animals

Inhibition of venous thrombogenesis by the sample drug was calculated by comparing the thrombogenicity of the control group administered with 10% HPCD (hydroxy propyl β-cyclodextrin) solution with the same volume as the sample volume at 1 mg / kg body weight. As shown in Table 2 below. Compound 1-5 of the present invention, a sample drug, showed an effective antithrombotic effect.

Experimental Example 3. Pharmacokinetics

The compounds of the present invention showed a high oral absorption tendency in animals. As one example, the results of the compound 1-5 of the present invention are shown in Tables 3 and 4 below. Table 3 shows the results obtained by absorbing the compound into the gastrointestinal tract and detecting high concentrations in the blood up to 2 hours after the administration of the compound to rats. Although the dose was 1/3 of the dose, the blood concentration peak of the absorbed drug showed similar pharmacokinetic properties as that of the rat.

As can be seen in the above experimental example, the thrombin inhibitor in which the aminopyridoaryl group according to the present invention is introduced has excellent inhibitory activity against thrombin, excellent selectivity to thrombin, and excellent oral absorption rate.

Claims (6)

Compounds represented by Formula 1, pharmaceutically acceptable salts, prodrugs, hydrates, solvates and isomers thereof: [Formula 1] Where Q is an aminopyridoaryl group, W is hydrogen, alkyl (C = 1-8), diarylmethyl (C = 6-10), arylalkyl (C = 5-12), heteroaryl (C = 4-10), heteroarylalkyl (C = 5-11), dicycloalkylmethyl (C = 3-6), or cycloalkyl (C = 3-8), X and Y are each independently hydrogen, alkyl (C = 1-8), cycloalkyl (C = 3-8), alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkyl Aminocarbonylalkyl, monoalkylaminocarbonylalkyl, aminocarbonylalkyl, methanesulfonyl, aminosulfonyl, and O-alkyloxalyl, n is 0, 1 or 2. The method of claim 1, Q is an aminopyridoaryl group, W is alkyl (C = 1-8), diarylmethyl (C = 6-10), arylalkyl (C = 5-12), heteroaryl (C = 4-10), heteroarylalkyl (C = 5--10 11) dicycloalkylmethyl (C = 3-6) or cycloalkyl (C = 3-8), X is hydrogen, alkyl (C = 1-8), cycloalkyl (C = 3-8), alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkylaminocarbonylalkyl, Monoalkylaminocarbonylalkyl, aminocarbonylalkyl, methanesulfonyl, aminosulfonyl, and O-alkyloxalyl, Y is hydrogen, alkyl (C = 1-8) or cycloalkyl (C = 3-8), n is 0, 1 or 2; The method of claim 2, Q is 1-amino-isoquinolin-6-yl, 1-amino-isoquinolin-7-yl, 1-amino-2-oxo-isoquinolin-6-yl or 7-amino-thieno [2,3c] Pyridin-2-yl, W is alkyl (C = 1-8), diarylmethyl (C = 6-10), arylalkyl (C = 5-12), heteroaryl (C = 4-10), heteroarylalkyl (C = 5--10 11) dicycloalkylmethyl (C = 3-6) or cycloalkyl (C = 3-8), X is hydrogen, alkyl (C = 1-8), cycloalkyl (C = 3-8), alkoxycarbonylalkyl, cyanoalkyl, carboxyalkyl, hydroxyalkyl, alkyloxyalkyl, dialkylaminocarbonylalkyl, Monoalkylaminocarbonylalkyl, aminocarbonylalkyl, methanesulfonyl, aminosulfonyl, and O-alkyloxalyl, Y is hydrogen or methyl, n is 0, 1 or 2; The method of claim 3, wherein 2- {2-[(1-Amino-isoquinolin-7-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid ethyl ester ; 2- {2-[(1-Amino-isoquinolin-7-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid ethyl ester ; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamino acetic acid; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- Aminosulfonyl) amines; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- Methanesulfonyl) amine; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- [O-methyloxalyl]) amine; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylamine; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- [Ethoxycarbonylmethanesulfonyl]) amine; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ¹ , N ¹ -dimethyl) amine; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl (N ^1- Cyanomethyl) amine; 2- {2-[(1-Amino-2-oxo-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethyl Amino acetic acid ethyl esters; 2- {2-[(1-Amino-isoquinolin-6-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) ethylaminosulfonyl acetic acid ; And 2- {2-[(7-aminothieno [2,3c] pyridin-2-ylmethyl) -carbamoyl] -pyrrolidin-1-yl} -2-oxo- (1-diphenylmethyl) A compound selected from the group consisting of ethylamino acetic acid ethyl ester. Preventing blood coagulation, comprising as an active ingredient a compound of formula 1 as defined in claim 1, a pharmaceutically acceptable salt, prodrug, hydrate, solvate or isomer together with a pharmaceutically acceptable carrier; and Pharmaceutical composition for the treatment of thrombosis. The composition of claim 5 formulated for oral administration.