KR19980032448A - Selective thrombin inhibitor that can be administered orally - Google Patents

Abstract

The present invention relates to a novel selective thrombin inhibitor having the following formula (1), which is effective even when used for oral administration, a process for producing the same, and a thrombin inhibitory pharmaceutical composition containing the compound as an active ingredient.

In this formula,

R ¹ and R ² are each independently hydrogen, alkyl or alkyloxy,

R ³ and R ⁴ each independently represent lower alkyl or cycloalkyl,

R ⁵ represents hydrogen, lower alkyl or amino.

Description

Selective thrombin inhibitor that can be administered orally

The present invention relates to a novel selective thrombin inhibitor having the following formula (1), which is effective even when used for oral administration, a process for producing the same, and a thrombin inhibitory pharmaceutical composition containing the compound as an active ingredient.

[Chemical Formula 1]

In this formula,

R ¹ and R ² are each independently hydrogen, alkyl or alkyloxy,

R ³ and R ⁴ each independently represent lower alkyl or cycloalkyl,

R ⁵ represents hydrogen, lower alkyl or amino.

It is generally known that various complex enzyme reactions are involved in the blood coagulation process. And the last step involves the conversion of prothrombin to thrombin. The thrombin generated in this process plays a role of activating the platelets and changing the fibrinogen to fibrin. The resulting fibrin is converted into a polymer substance by the polymerization reaction, cross-linked by the activated blood factor XIII, . Thrombin also activates blood factors V and VIII that participate in the blood clotting process, further accelerating the blood clotting reaction. Therefore, the thrombin inhibitor acts as an effective anticoagulant and inhibits platelet activity and prevents fibrinogenesis and stabilization. Therefore, by developing a new substance capable of inhibiting thrombin activity for a long time, blood coagulation is prevented and various thrombosis A method for treatment has been sought.

However, the ability to simply inhibit thrombin is limited by its use as an effective anticoagulant. This is because thrombin is a serine protease similar to trypsin, and effective thrombin inhibitors have a high inhibitory effect on trypsin. Since the serine protease (representative example: plasmin), which is similar to trypsin, is present in the human body, especially in the blood, the serine protease in the development of the thrombin inhibitor, It is very important to have the property that

Under these circumstances, extensive research has been conducted to develop an selective thrombin inhibitor that effectively inhibits thrombin and low inhibitory activity against trypsin.

Representative compounds that have been developed as effective thrombin inhibitors include Argatroban, which is the first arylsulfonylarginine compound (Chemical Formula 10): (US Pat. Nos. 4258192 and 4201863).

This compound has been reported to inhibit thrombin by 250-fold better than trypsin, and has already been commercialized in Japan as a injectable preparation in 1990 (Biochemistry 1984, 23, pp85-90). However, this compound has no disadvantage in that it is not active for oral administration and is synthesized through a very complicated process.

NAPAP, which is a benzamidine-based arylsulfonyl compound, was also developed as a thrombin inhibitor. However, this compound is not only easy to synthesize but also is an effective thrombin inhibitor. However, the thrombin inhibitory effect against trypsin is only about 50 times (See J. Biol. Chem. 1991, 266, pp20085-20093).

On the other hand, a compound of Ro 46-6240 of the following formula (12) with improved selectivity against trypsin has been reported as a powerful thrombin inhibitor. However, the compound has potential to develop as a preparation for intravenous use, (See J. Med. Chem. 1994, 37, pp3889-3901].

As a representative compound that has been developed as an effective thrombin inhibitor capable of oral administration, there is CVS-1123 of the following chemical formula 13 developed by Corvas. This compound has been reported to be able to be administered orally to dogs and monkeys as well as to rats, but it is a disadvantage that the selectivity to trypsin is poor (WO 9315756, WO 9408941).

Thus, the present inventors have conducted intensive studies for a long period of time to develop compounds capable of orally administering thrombin with a selectivity to tryptin being remarkably improved while exhibiting relatively easy and effective thrombin inhibitory activity, The inventors of the present invention confirmed that the compound of the formula (1) according to the present invention can achieve this object effectively, and completed the present invention.

Accordingly, an object of the present invention is to provide a novel thrombin inhibitor which is orally administrable and has high selectivity, and a method for producing the same.

The present invention also relates to a composition for the prevention of blood clotting or for the treatment of various thrombosis, which comprises the above compound as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention relates to a compound represented by the following general formula (1), a pharmaceutically acceptable salt thereof and an isomer thereof.

[Chemical Formula 1]

In this formula,

R ¹ and R ² are each independently hydrogen, alkyl or alkyloxy,

R ³ and R ⁴ each independently represent lower alkyl or cycloalkyl,

R ⁵ represents hydrogen, lower alkyl or amino.

In the above definitions of substituents of the compounds of formula (I) according to the invention, the term alkyl, when used alone or in combination with alkyloxy, means methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, Quot; lower alkyl " means straight or branched chain hydrocarbon radicals having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, butyl, or isomers thereof, The term cycloalkyl means cyclic alkyl of 3 to 8 carbon atoms including cyclopentyl.

Preferred compounds of formula (I) of the present invention are those wherein R ¹ and R ² are each independently hydrogen, C ₁ -C ₅ alkyl or C ₁ -C ₃ alkyloxy, R ³ and R ⁴ are each independently C ₁ -C ₃ alkyl or C ₃ -C ₅ cyclic alkyl, and R ⁵ represents C ₁ -C ₃ alkyl or amino.

Particularly preferred compounds of formula (I) of the present invention are those wherein R ¹ and R ² are each independently hydrogen, methyl, ethyl, propyl, butyl or isomer thereof, pentyl or isomer thereof or ethyloxy, R ³ and R ⁴ are each independently Methyl, ethyl or cyclopentyl, and R ⁵ is methyl or amino.

Representative examples of compounds of formula (I) according to the present invention include the following compounds:

1. Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl- 2- (4-propyl-benzenesulfonylamino) propionamide

2. Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl- 2- (4-butyl- benzenesulfonylamino) propionamide

3. Synthesis of (S) -3- (4-amidorazo-phenyl) -N-cyclopentyl-N-methyl- 2- (4- tert- butyl- benzenesulfonylamino) propionamide

4. Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl- 2- (3,4- diethyl- benzenesulfonylamino) propionamide

5. Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2-

6. Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-isoamyl-benzenesulfonylamino) propionamide

7. (S) -3- (4-Amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4- ethyl- benzenesulfonylamino) propionamide

8. Preparation of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (3-

9. Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-ethyl- 2- (4-isobutyl-benzenesulfonylamino) propionamide

10. Synthesis of (S) -3- [4- (amino-methylimino-methyl) -phenyl] -N-cyclopentyl-N-methyl- 2- (4-propyl-benzenesulfonylamino) propionamide

11. Preparation of (S) -3- (4-amidorazo-phenyl) -N-cyclopentyl-2- (4-isobutyl- benzenesulfonylamino)

12. Preparation of (S) -3- [4- (amino-methylimino-methyl) -phenyl] -N-cyclopentyl-2- (4-isobutyl-benzenesulfonylamino)

13. Preparation of (S) -3- (4-amidazo no-phenyl) -N-cyclopentyl-2- (2-isobutyl-benzenesulfonylamino)

14. Preparation of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-2- (4- (2,2- dimethyl- propyl) -benzenesulfonylamino)

15. A process for the preparation of (S) -3- [4- (amino-methylimino-methyl) -phenyl] -N-cyclopentyl-2- (4- (2,2- N-methyl-propionamide

16. Preparation of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl- 2- (4-ethyloxy- benzenesulfonylamino)

The compounds of formula (I) according to the invention may also form pharmaceutically acceptable salts. Such pharmaceutically acceptable salts include those acids which form pharmaceutically acceptable anions and which form non-toxic acid addition salts such as hydrochloric acid, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrobromic acid, hydroiodic acid, tartaric acid, formic acid, Organic carboxylic acids such as acetic acid, acetic acid, trichloroacetic acid or trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid and the like, sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid And the like.

On the other hand, the compounds according to the present invention may have an asymmetric carbon center, and thus may exist as racemates, diastereomer mixtures and individual diastereomers, all of which are included in the scope of the present invention.

The present invention also relates to a process for the preparation of the compounds of formula I as defined above.

According to the present invention, the compound of formula 1 defined above can be prepared by reacting a methylmercapto compound of formula 2 with an amine derivative of formula 3:

The process for preparing the compound of formula (1) according to the present invention can be illustrated by the following reaction formula (1).

Wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined above.

As shown in Reaction Scheme 1, the compound of Formula 1 according to the present invention can be prepared by reacting the methylmercapto compound of Formula 2 with an amine derivative of Formula 3, which is a nucleophile.

This reaction can be preferably carried out in the presence of a solvent. Examples of the solvent that can be preferably used for this purpose include any organic solvent which does not adversely affect the reaction, but alcohol solvents such as methanol, ethanol, propanol and the like are preferably used.

The reaction conditions including the used amount of the reactants, the reaction temperature, the reaction time, and the like in the present reaction can be easily determined by those skilled in the art depending on the specific reactants. In general, the reaction temperature can be varied, but it is particularly preferable to carry out the reaction at 0 캜 to 50 캜. The reaction time is generally 0.5 to 5 hours, but preferably the reaction is carried out for 1 to 2 hours.

After completion of the reaction, the product can be separated and purified by a conventional post-treatment method such as chromatography or recrystallization.

The methylmercapto compound of formula 2 used as an intermediate in the preparation of the compound of formula 1 in Scheme 1 may be prepared by coupling a compound of formula 4, for example, with an amine substituted with R ³ and R ⁴ to give a compound of formula 5 , Removing the amino protecting group from the compound of formula (5), reacting the resulting compound of formula (6) with the sulfonyl derivative of formula (7) to introduce the sulfonyl group at the N- Is saturated with hydrogen sulfide in the presence of a base to produce a thioamide compound of formula (9), and methylating the resulting compound of formula (9).

Wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined above and P is a group such as benzyloxycarbonyl, allyloxycarbonyl, t-butoxycarbonyl or trityl groups Amino protecting group.

The method for producing the compound of formula (2) as described above can be represented by the following reaction formula (2).

As shown in Scheme 2, to prepare the compound of Formula 2, the amine group substituted with R ³ and R ⁴ at the C-terminal of the compound of Formula 4 is first coupled to obtain the compound of Formula 5.

Known coupling reagents that can be used for the coupling reaction of the compound of Formula 4 include dicyclohexylcarbodiimide (DCC), 3-ethyl-3'- (dimethylamino) -propylcarbodiimide (EDC) (2-oxo-3-oxazolidinyl) -phosphinic acid chloride (BOP-Cl), diphenylphosphoryl azide (DPPA), and the like.

The carboxylic acid compound of formula (4) used in this coupling reaction may be used as it is, but is preferably converted into its reactive derivative, for example, acid halide and other activated ester derivative, . Activated derivatives of carboxylic acids are necessary to form amide bonds by coupling reactions with amines or to form ester bonds by coupling reactions with alcohols. Such reactive derivatives include conventional derivatives that can be prepared by methods conventional in the art, for example, acid halides include acid chlorides, and activated esters include methoxycarbonyl chloride, isobutyloxycarbonyl N-hydroxyphthalimide-derived esters, N-hydroxysuccinimide-derived esters, N-hydroxy-5- < RTI ID = 0.0 > Norbornene-2 ', 3'-dicarboxyimide-derived esters, 2,4,5-trichlorophenol-derived esters, and the like.

The compound of formula 5 produced by the coupling reaction of the compound of formula 4 with an amine group is then removed by conventional methods to obtain the compound of formula 6 by removing the N-terminal amino protecting group. Reacting the compound of formula (6) with a sulfonyl chloride compound of formula (7) to introduce a sulfonyl group into the N-terminal part of the compound to obtain a compound of formula (8), then reacting the nitrile compound of formula (8) with pyridine and trimethylamine Saturated with hydrogen sulfide in the presence of the same base to give the thioamide compound of formula (9), followed by subsequent methylation of the thioamide compound of formula (9) to give the desired compound of formula (2). A methylating agent such as iodomethane, dimethyl sulfate ((CH ₃ ) ₂ SO ₂ ) or methyl triflate (CH ₃ OTf) may be used to methylate the compound of formula (9).

As described above, the novel compounds of formula (I) according to the present invention are more selective than known compounds and exhibit an effective thrombin inhibitory effect by oral administration. Accordingly, the compounds of the present invention are useful for the prevention of blood clotting and the treatment of thrombosis.

Accordingly, another object of the present invention is to provide a pharmaceutical composition for the prevention of blood clotting and the treatment of thrombosis, which comprises a compound of the formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

The total daily dose to be administered to a host in single or divided doses upon administration of a compound of the present invention for clinical purposes is preferably in the range of 0.001 mg to 10 mg per kilogram of body weight, The severity of the disease, the particular compound being administered, the weight, sex, health condition, diet, time of administration, method of administration, excretion rate, drug mix and severity of the individual patient.

The compounds of the present invention may be administered as injectable preparations and oral preparations as desired.

Injectable preparations such as sterile injectable aqueous or oleaginous suspensions may be prepared using suitable dispersing agents, wetting agents or suspending agents according to known techniques. Pharmaceutically acceptable solvents that may be used include water, Ringer's solution and isotonic NaCl solution, and sterile, fixed oils are typically used as a solvent or suspending medium. Any non-irritating fixed oils, including mono-, di-glycerides, may be used for this purpose, and fatty acids such as oleic acid are also used in the injectable preparations.

Solid dosage forms for oral administration may be in the form of capsules, tablets, pills, powders and granules, with capsule and tablet being particularly useful. Tablets and pills are preferably prepared as preservative. Solid dosage forms can be prepared by admixing the active compound of Formula 1 according to the present invention with one or more inert diluents such as sucrose, lactose, starch and the like and a carrier such as a lubricant such as magnesium stearate, disintegrant, binder, and the like.

One of the great features of the compound of formula (I) according to the present invention is that it shows excellent pharmacological effects when it is formulated into an oral formulation and then orally administered. As a result of pharmacokinetic experiment, it was proved that when the pharmaceutical composition of the present invention is orally administered, the concentration of the drug is maintained in the blood for a long time. Therefore, unlike conventional thrombin inhibitors, it is more useful in that it can be effectively used as an oral preparation.

In addition, in the course of performing pharmacokinetic experiments, it has been confirmed that the compounds according to the present invention do not show acute toxicity to mammals such as rats and dogs and achieve the desired scavenging purpose.

When the compound of the present invention is to be clinically administered to obtain the desired anticoagulant effect and thrombolytic effect, the active compound of the formula (1) according to the present invention may be used as a thrombolytic agent and / Can be administered at the same time. Examples of thrombolytic agents that can be administered in combination with the compound of the present invention in this manner include t-PA, urokinase, streptokinase, and the like. Examples of the thrombocyte activity inhibitor include aspirin, ticlopidine, Clopidrogel, 7E3 monoclonal antibody, and the like.

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

The present invention will be described more specifically by the following examples and experimental examples, but the scope of the present invention is not limited in any way by them.

Preparation Example 1: Synthesis of cyclopentyl-methylamine

10 ml (113 mmol) of cyclopentanone was dissolved in 50 ml of methanol, and 50 ml of water was added thereto. 7.6 g (113 mmol) of methylamine hydrochloride was added to the reaction mixture, and 7.1 g (113 mmol) of sodium borocyanohydride (NaBH ₃ CN) was added thereto, followed by stirring under reflux under a condition of pH 6 for 12 hours . The reaction solution was cooled to 0 ° C and the pH was adjusted to neutral with 6N sodium hydroxide solution. The methanol was removed under reduced pressure, cooled to 0 < 0 > C, adjusted to pH 2 with dilute hydrochloric acid and washed three times with diethyl ether. The aqueous layer was adjusted to pH 11 again. 50 ml of dioxane and 37 g (169.5 mmol) of butyloxycarbonyl anhydride were added thereto, and the mixture was stirred at room temperature for 3 hours. After the reaction was completed, the reaction solution was distilled under reduced pressure, concentrated to a volume of about 30 ml, extracted with ethyl acetate, and washed with an aqueous acid and base solution. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated. The resulting white solid was purified by column chromatography [eluent: ethyl acetate / hexane (7: 3, v / v)]. The obtained solid was dissolved in 60 ml of a 4N hydrochloric acid-dioxane solution and stirred for 30 minutes. Then, the solvent was removed under reduced pressure and vacuum dried to obtain 13.7 g (yield: 90.5%) of the title compound.

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 3.50 (m, 1H), 2.68 (s, 3H), 2.10 (m, 2H), 1.86-1.50 (m, 6H)

Preparation Example 2 Synthesis of (S) -3- (4-cyanophenyl) -N-cyclopentyl-2- (butyloxycarbonylamino) -N-methyl-propionamide

(2.41 mmol) of (S) -3- (4-cyanophenyl) -2- (butyloxycarbonylamino) propionic acid were dissolved in 6 ml of dimethylformamide (DMF) To this solution, 0.7 g of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC) and 0.4 g of 1-hydroxybenzotriazole hydrate (HOBT) were added and stirred until completely dissolved. 0.4 g (2.96 mmol) of the compound obtained in Preparation Example 1 and 1.0 ml of N-methylmorpholine were added to the solution, the reaction temperature was gradually raised to room temperature, and the mixture was stirred for 3.5 hours. After completion of the reaction, the reaction solution was distilled under reduced pressure to remove volatile substances. The resulting residue was diluted with ethyl acetate, and washed successively with a saturated aqueous sodium hydrogen carbonate solution, diluted hydrochloric acid and saturated brine. The residue was purified by column chromatography [eluent: ethyl acetate / hexane (7: 3, v / v)] to obtain 0.65 g of the purified title compound (yield: 73.0%).

_{^{1 H NMR (CDCl 3, ppm}} ) δ: 7.61 (m, 2H), 7.32 (m, 2H), 5.48, 5.01-4.86, 4.12 (m, m, m, 3H), 2.75, 2.62 (s, s, 3H), 2.90-1.20 (m, 17H)

Mass (FAB, m / e): 372 (M ^< + & gt ^; +1)

Preparation 3: Synthesis of 4-propyl-benzenesulfonyl chloride

1.5 ml (22.6 mmol) of chlorosulfonic acid were added to the reaction vessel and cooled to 0 占 폚. 1.0 ml (7.17 mmol) of propylbenzene was slowly added dropwise while care was taken so that the temperature did not rise above 5 占 폚. After the dropwise addition was completed, the reaction solution was raised to room temperature and stirred for 3 hours. The reaction solution was cooled again to 0 < 0 > C and ice was added to terminate the reaction. 40 ml of water and ethyl acetate were added to the reaction mixture three times, and the mixture was extracted three times. The organic layers were combined, dried over anhydrous magnesium sulfate, filtered and the filtrate was concentrated to obtain 1.3 g of a solution. The obtained compound was a mixture of 2-propylbenzenesulfonyl chloride and 4-propylbenzenesulfonyl chloride, and the ratio thereof by NMR analysis was 1: 2. The yield of the title compound was 0.86 g (Yield: 55%).

_{^{1 H NMR (CDCl 3, ppm}} ) δ: 8.04 (dd, 2H), 7.49 (dd, 2H), 2.82 (t, 2H), 1.78 (m, 2H), 1.06 (t, 3H)

Mass (FAB, m / e): 219 (M ^< + & gt ^; +1)

Preparation Example 4: Synthesis of 3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (4-propyl-benzenesulfonylamino)

0.97 g (2.61 mmol) of the compound obtained in Preparation Example 2 was dissolved in 10 ml of methanol and then cooled to 0 占 폚. An excess of acetyl chloride (30 equivalents) was slowly added thereto, and the mixture was stirred at room temperature for 1 hour. All volatiles were removed under reduced pressure to give a white solid, which was dried in vacuo and dissolved in 10 ml of DMF. 1.0 ml of N-methylmorpholine was added thereto, and the mixture was stirred for 10 minutes. To the reaction mixture was added 0.7 g (3.21 mmol) of the mixture obtained in Preparation Example 3 and stirred for 1 hour. The solvent was removed under reduced pressure and 40 ml of water and dichloromethane were added three times. The organic layers were combined, dried over anhydrous magnesium sulfate, filtered and the filtrate was concentrated. The obtained solid was separated and purified by column chromatography [eluent: hexane / ethyl acetate (7: 3, v / v)] to obtain 0.8 g (yield: 82%) of the title compound.

_{^{1 H NMR (CDCl 3, ppm}} ) δ: 7.72 (dd, 2H), 7.65 (m, 2H), 7.34 (m, 4H), 5.86 (m, 1H), 4.63, 4.56, 4.39, 3.88 (m, m (m, 2H), 2.69 (m, 2H), 2.59, 2.40 (s,

Mass (FAB, m / e): 454 (M ^< + & gt ^; +1)

Example 1: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-propyl-benzenesulfonylamino) propionamide

0.45 g (0.99 mmol) of the compound obtained in Preparation Example 4 was dissolved in 10 ml of pyridine, placed in a branched flask, and 1.0 ml of triethylamine was added thereto. From one branch of the flask, hydrogen sulphide (H ₂ S) gas was slowly flowed into the solution and the gas was flowed out from the other branch. The hydrogen sulfide gas was saturated with the solution while stirring for about 10 minutes. At this time, the color of the solution changed from colorless to greenish and gradually changed to dark brown. The flask was closed with a rubber stopper, and the reaction was completed by allowing it to stand at room temperature for 3 days. Subsequently, volatile substances were removed by distillation under reduced pressure, and the reaction product was dried with a vacuum pump. 10 ml of acetonitrile and 0.25 ml of methane iodide (CH ₃ I) were added to the obtained yellow solid, and the mixture was heated under reflux for 30 minutes. The volatiles were removed by distillation under reduced pressure, and the residue was dried with a vacuum pump. The residue was dissolved in 8 ml of anhydrous methanol and stirred. To this was added 0.04 ml (0.66 mmol) of 80% hydrazine hydrate (H ₂ NNH ₂ .H ₂ O) three times at 10 minute intervals. After the reaction was completed, the reaction solution was concentrated, and the residue was purified by column chromatography [eluent: dichloromethane / methanol (95: 5, v / v)] to obtain 0.19 g of the title compound (yield: 80% Respectively.

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 7.66 (m, 4H), 7.42 (m, 2H), 7.33 (m, 2H), 4.58, 4.43, 4.05 (m, m, m, 2H), 3.04, (M, 2H), 2.66 (m, 2H), 2.59 (s, 3H)

Mass (FAB, m / e): 486 (M ^< + & gt ^; +1)

Preparation Example 5: Synthesis of 4-butyl-benzenesulfonyl chloride

The reaction was carried out in the same manner as in Production Example 3, except that butylbenzene was used instead of propylbenzene to obtain 0.6 g (yield: 50%) of the title compound.

Preparation Example 6: Synthesis of 3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (4-butylbenzenesulfonylamino)

The reaction was carried out in the same manner as in Production Example 4 using the compound obtained in Preparation Example 5 to obtain 0.5 g (yield: 80%) of the title compound.

_{^{1 H NMR (CDCl 3, ppm}} ) δ: 7.2-7.7 (m, 8H), 5.7 (m, 1H), 4.6 (m, 0.5H), 4.5 (m, 0.5H), 4.4 (m, 0.5H) , 3.8 (m, 0.5H), 2.9-3.0 (m, 2H), 2.6-2.7 (m, 2H), 2.5 (s, 1.4H) )

Mass (FAB, m / e): 468 (M ^< + & gt ^; +1)

Example 2: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-butyl- benzenesulfonylamino) -propionamide

The reaction was carried out in the same manner as in Example 1 using the compound obtained in Preparation Example 6 to obtain 0.11 g (yield: 40%) of the title compound.

¹ H NMR (CD ₃ OD, ppm)?: 7.4-7.8 (m, 8H), 4.7 (m, 0.5H), 4.5-4.6 (m, 2H), 2.8 (m, 2H), 2.7 (s, 1.6H)

Mass (FAB, m / e): 500 (M ^< + & gt ^; +1)

Preparation 7: Synthesis of 3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (4-t- butyl- benzenesulfonylamino) -propionamide

The reaction was carried out in the same manner as in Production Example 4, except that 4-t-butylbenzenesulfonyl chloride was used instead of 4-propyl-benzenesulfonyl chloride to give 0.52 g (yield: 82%) of the title compound Respectively.

_{^{1 H NMR (CDCl 3, ppm}} ) δ: 7.2-7.8 (m, 8H), 5.8 (m, 1H), 4.6 (m, 0.5H), 4.5 (m, 0.5H), 4.4 (m, 0.5H) 2H), 2.5 (s, 1.3H), 2.3 (s, 1.7H), 0.9-1.7 (m, 17H)

Mass (FAB, m / e): 468 (M ^< + & gt ^; +1)

Example 3: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4- tert- butyl- benzenesulfonylamino) -propionamide

The reaction was carried out in the same manner as in Example 1 using the compound obtained in Preparation Example 7 to obtain 0.12 g (yield: 42%) of the title compound.

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 7.5-7.9 (m, 8H), 4.7 (m, 0.5H), 4.6 (m, 0.5H), 4.4 (m, 0.5H), 4.2 (m, 0.5 2H), 2.7 (s, 1.5H), 2.6 (s, 1.5H), 1.2-1.9 (m, 17H)

Mass (FAB, m / e): 500 (M ^< + & gt ^; +1)

Preparation Example 8: Synthesis of 3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (3,4-diethyl- benzenesulfonylamino)

(Yield: 83%) was prepared in the same manner as in Production Example 4, except that 3,4-diethyl-benzenesulfonyl chloride was used in place of 4-propyl-benzenesulfonyl chloride. ≪ / RTI >

_{^{1 H NMR (CDCl 3, ppm}} ) δ: 7.1-7.6 (m, 7H), 5.8 (m, 1H), 4.4-4.7 (m, 1H), 4.4 (m, 0.5H), 3.7 (m, 0.5H ), 2.9-3.1 (m, 2H), 2.7 (m, 4H), 2.6 (s, 1.4H)

Mass (FAB, m / e): 468 (M ^< + & gt ^; +1)

Example 4: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl- 2- (3,4- diethyl- benzenesulfonylamino) - propionamide

The reaction was carried out in the same manner as in Example 1 using the compound obtained in Preparation Example 8 to obtain 0.13 g (yield: 44%) of the title compound.

¹ H NMR (CD ₃ OD, ppm)?: 7.3-7.8 (m, 7H), 4.6 (m, 0.5H), 4.5 2H), 2.7-2.8 (m, 4H), 2.6 (s, 1.6H)

Mass (FAB, m / e): 500 (M ^< + & gt ^; +1)

Preparation Example 9: Synthesis of 3-methyl-4-propyl-benzenesulfonyl chloride

The reaction was carried out in the same manner as in Production Example 3, except that 2-propyltoluene was used instead of propylbenzene to obtain 0.7 g (Yield: 40%) of the title compound.

Preparation 10: Synthesis of 3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (3-methyl-4-propyl-benzenesulfonylamino)

The reaction was carried out according to the same method as in Production Example 4 using the compound obtained in Production Example 9 to obtain 0.4 g (yield: 65%) of the title compound.

¹ H NMR (CDCl ₃ , ppm)?: 7.1-7.6 (m, 7H), 5.8 (m, 1H), 4.5-4.7 ), 2.9-3.1 (m, 2H), 2.6-2.7 (m, 4H), 2.3-2.4 (s, 6H)

Mass (FAB, m / e): 468 (M ^< + & gt ^; +1)

Example 5: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2-

The reaction was carried out in the same manner as in Example 1 using the compound obtained in Preparation Example 10 to obtain 0.09 g (yield: 43%) of the title compound.

¹ H NMR (CD ₃ OD, ppm)?: 7.3-7.8 (m, 7H), 4.7 (m, 0.5H), 4.5 2H), 2.7 (m, 2H), 2.6 (s, 1.5H), 2.5 (s, 1.5H)

Mass (FAB, m / e): 500 (M ^< + & gt ^; +1)

Preparation 11: Synthesis of 4-isoamyl-benzenesulfonyl chloride

The reaction was carried out in the same manner as in Production Example 3, except that isoamylbenzene was used instead of propylbenzene to obtain 1.43 g (yield: 86%) of the title compound.

Example 6: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-isoamyl-benzenesulfonylamino)

(S) -3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (4 0.35 g (1.25 mmol) of the compound obtained in the previous step was reacted according to the same procedure as in Example 1 to give the title compound (0.45 g, Yield: 71%).

¹ H NMR (CD ₃ OD, ppm)?: 7.4-7.8 (m, 8H), 4.7 (m, 0.5H), 4.5-4.6 (m, 2H), 2.8 (m, 2H), 2.7 (s, 1.6H)

Mass (FAB, m / e): 514 (M ^< + & gt ^; +1)

Preparation 12: Synthesis of 4-ethyl-benzenesulfonyl chloride

The reaction was carried out in the same manner as in Production Example 3, except that ethylbenzene was used instead of propylbenzene to obtain 0.84 g (yield: 81%) of the title compound.

Example 7: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-ethyl-benzenesulfonylamino)

(S) -3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (4 (0.93 mmol) of 2-ethoxy-benzenesulfonylamino) -propionamide, which was reacted according to the same procedure as in Example 1 to give the title compound 0.18 g (yield: 41%).

¹ H NMR (CD ₃ OD, ppm)?: 7.4-7.8 (m, 8H), 4.65 (m, 0.5H), 4.45-4.55 2H), 2.65 (s, 1.6H), 2.55 (s, 1.4H), 0.9 - 1.8 (m,

Mass (FAB, m / e): 472 (M ^< + & gt ^; +1)

Preparation 13: Synthesis of 3-methyl-4-ethyl-benzenesulfonyl chloride

The reaction was carried out in the same manner as in Production Example 3, except that ethylbenzene was used instead of propylbenzene to obtain 3.2 g (yield: 99%) of a mixture of the title compound and 4-methyl-3-ethyl-benzenesulfonyl chloride, ≪ / RTI >

Example 8: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (3-methyl-3-ethyl-benzenesulfonylamino)

(S) -3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (3 (4-methyl-3-ethyl-benzenesulfonylamino) -propionamide and 3- (4-cyanophenyl) -N-cyclopentyl- ) -Propionamide (0.5 g, 1.10 mmol) was obtained and reacted according to the same procedure as in Example 1 to give 0.35 g (yield: 65%) of a mixture of the title compound and the positional isomer.

¹ H NMR (CD ₃ OD, ppm)?: 7.3-7.75 (m, 7H), 4.65 (m, 0.5H), 4.45-4.55 m, 2H), 2.75 (m, 2H), 2.64, 2.55 (d,

Mass (FAB, m / e): 486 (M ^< + & gt ^; +1)

Preparation Example 14: Synthesis of cyclopentyl-ethylamine hydrochloride

The reaction was carried out according to the same procedure as in Preparation Example 1, except that ethylamine hydrochloride was used instead of methylamine hydrochloride to obtain 3.0 g (Yield: 88%) of the title compound.

Preparation 15: Synthesis of (S) -3- (4-cyanophenyl) -N-cyclopentyl-2- (butyloxycarbonylamino) - N-ethyl-propionamide

The reaction was carried out in the same manner as in PREPARATION 2 using the compound obtained in PREPARATION 14 instead of cyclopentyl-methylamine hydrochloride to obtain 0.92 g (Yield: 36%) of the title compound.

¹ H NMR (CDCl ₃ , ppm)?: 7.3-7.75 (m, 4H), 5.6-5.4 (m, 1H), 4.9 H), 4.05 (m, 0.5H), 3.3-2.95 (m, 2H), 2.1-0.9

Mass (FAB, m / e): 386 (M ^< + & gt ^; +1)

Preparation 16: Synthesis of 4-isobutyl-benzenesulfonyl chloride

The reaction was carried out in the same manner as in Production Example 3, except that 4-isobutylbenzene was used instead of propylbenzene to obtain the title compound (yield: 86%).

Example 9: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-ethyl-2- (4-isobutyl-benzenesulfonylamino)

The reaction was carried out in the same manner as in Production Example 4 using the compound obtained in Preparation Example 15 and the compound obtained in Preparation Example 16 to obtain the intermediate (S) -3- (4-cyanophenyl) -N-cyclopentyl- (0.95 mmol) of N-ethyl-2- (4-isobutyl-benzenesulfonylamino) -propionamide was obtained and reacted according to the same procedure as in Example 1 to give the title compound 0.35 g (yield: 72% ).

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 7.25-7.82 (m, 8H), 4.65 (m, 0.5H), 4.4 (m, 0.5H), 4.18 (m, 0.5H), 4.05 (m, 0.5 (M, 4H), 2.61 (m, 2H), 1.25-1.8 (m, 8H), 0.9-1.1

Mass (FAB, m / e): 514 (M ^< + & gt ^; +1)

Example 10: Synthesis of (S) -3- [4- (amino-methylimino-methyl) -phenyl] -N-cyclopentyl-N-methyl- 2- (4-propyl-benzenesulfonylamino) -propionamide Synthesis of

The procedure of Example 1 was repeated except for using a 2.0 M methylamine solution dissolved in methanol instead of hydrazine hydrate to give 68 mg (yield: 28%) of the title compound.

¹ H NMR (CD ₃ OD, ppm)?: 7.66 (m, 4H), 7.43 (m, 2H), 7.33 (m, 2H), 4.59,4.43,4.09 (s, 3H), 3.05, 2.89 (m, 2H), 2.65 (m, 2H), 2.60

Mass (FAB, m / e): 485 (M ^< + & gt ^; +1)

Example 11: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-2- (4-isobutyl-benzenesulfonylamino) -N-methyl-propionamide

The reaction was carried out in the same manner as in Production Example 3 except that isobutylbenzene was used instead of propylbenzene to synthesize 4-isobutyl-benzenesulfonyl chloride, which was synthesized in Production Example 4 using 4-propyl-benzenesulfonyl chloride 3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (4-isobutyl- benzenesulfonylamino) - propionamide in 58% ≪ / RTI > Using the procedure described in Example 1, the title compound (0.11 g, yield: 96%) was obtained.

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 7.65 (m, 4H), 7.44 (m, 2H), 7.32 (m, 2H), 4.61, 4.45, 4.09 (m, m, m, 2H), 3.04, 2H), 1.74 (m, 2H), 1.94-1.10 (m, 8H), 0.91 (d, 6H)

Mass (FAB, m / e): 500 (M ^< + & gt ^; +1)

Example 12: Preparation of (S) -3 - [(4-amino-methylimino-methyl) -phenyl] -N-cyclopentyl-2- (4-isobutyl-benzenesulfonylamino) Synthesis of amide

The reaction was carried out in the same manner as in Example 11, except that a 2.0 M methylamine solution dissolved in methanol instead of hydrazine hydrate was used to obtain 87 mg (Yield: 76%) of the title compound.

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 7.65 (m, 4H), 7.39 (m, 2H), 7.27 (m, 2H), 4.59, 4.41, 4.08 (m, m, m, 2H), 3.08 ( (s, 3H), 3.03, 2.85 (m, 2H), 2.58, 2.42 (s, 3H), 2.49 0.87 (d, 6H)

Mass (FAB, m / e): 499 (M ^< + & gt ^; +1)

Example 13: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-2- (2-isobutyl-benzenesulfonylamino) -N-methyl-propionamide

2-isobutyl-benzenesulfonyl chloride was synthesized in the same manner as in Production Example 3, except that isobutylbenzene was used instead of propylbenzene, to give 4-propyl-benzenesulfonyl chloride in Production Example 4, (4-cyanophenyl) -N-cyclopentyl-2- (2-isobutyl-benzenesulfonylamino) -N-methyl-propionamide in 7.1% ≪ / RTI > Using the procedure described in Example 1, the title compound (0.34 g, yield: 79%) was obtained.

¹ H NMR (CD ₃ OD, ppm)?: 7.96 (m, 1H), 7.72 (m, 2H), 7.70-7.25 (m, 5H), 4.62, 4.52, 4.46, (M, 2H), 3.18-2.73 (m, 4H), 2.57 (s, 3H), 2.12

Mass (FAB, m / e): 500 (M ^< + & gt ^; +1)

Example 14: Preparation of (S) -3- (4-amidorazo-phenyl) -N-cyclopentyl-2- (4- (2,2- dimethyl- propyl) -benzenesulfonylamino) Synthesis of propionamide

The reaction was carried out in the same manner as in Production Example 3 except that 2,2-dimethyl-propylbenzene was used instead of propylbenzene to synthesize 4- (2,2-dimethyl-propyl) -benzenesulfonyl chloride (S) -3- (4-cyanophenyl) -N-cyclopentyl-2- (4- (2,2-dimethyl- Propyl) -benzenesulfonylamino) -N-methyl-propionamide in 54% yield. Using the procedure described in Example 1, the title compound (0.13 g, yield: 93%) was obtained.

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 7.65 (m, 4H), 7.43 (m, 2H), 7.29 (m, 2H), 4.61, 4.45, 4.10 (m, m, m, 2H), 3.08, (S, 3H), 2.57 (m, 2H), 1.80-1.10 (m, 8H), 0.89

Mass (FAB, m / e): 514 (M ^< + & gt ^; +1)

Example 15: (S) -3- [4- (Amino-methylimino-methyl) -phenyl] -N-cyclopentyl-2- (4- (2,2- dimethyl- propyl) -benzenesulfonylamino ) -N-methyl-propionamide

The reaction was carried out in the same manner as in Example 14 except that a 2.0 M methylamine solution dissolved in methanol was used instead of hydrazine hydrate to obtain 0.23 g (yield: 82%) of the title compound.

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 7.68 (m, 4H), 7.45 (m, 2H), 7.30 (m, 2H), 4.61, 4.47, 4.12 (m, m, m, 2H), 3.12 ( 3H), 2.58 (m, 2H), 1.80-1.10 (m, 8H), 0.95 (s, 9H)

Mass (FAB, m / e): 513 (M ^< + & gt ^; +1)

Preparation 17: Synthesis of 4-ethyloxy-benzenesulfonyl chloride

The reaction was carried out in the same manner as in Production Example 3, except that 4-ethyloxybenzene was used instead of propylbenzene to obtain the title compound (yield: 65%).

Preparation 18: Synthesis of 3- (4-cyanophenyl) -N-cyclopentyl-N-methyl-2- (4-ethyloxy- benzenesulfonylamino) -propionamide

The reaction was carried out in the same manner as in Production Example 4 using the compound obtained in Production Example 17 to obtain 0.5 g (yield: 72%) of the title compound.

_{^{1 H NMR (CDCl 3, ppm}} ) δ: 7.6 (m, 4H), 7.4 (d, 2H), 6.9 (d, 2H), 5.9 (s, 1H), 4.0-4.7 (m, 4H), 3.1 ( m, 2H), 2.6 (s, 1.6H), 2.5 (s, 1.4H), 1.1-1.9

Mass (FAB, m / e): 455 (M ^< + & gt ^; +1)

Example 16: Synthesis of (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-ethyloxy- benzenesulfonylamino) -propionamide

The reaction was carried out in the same manner as in Example 1, using the compound obtained in Preparation 18, to give 0.19 g (yield: 43%) of the title compound.

_{^{1 H NMR (CD 3 OD,}} ppm) δ: 7.6-7.8 (m, 4H), 7.5 (d, 2H), 6.9 (d, 2H), 4.1-4.7 (m, 4H), 2.9-3.1 (m, 2H), 2.65 (s, 1.6H), 2.6 (s, 1.4H), 1.2 - 1.8 (m,

Mass (FAB, m / e): 487 (M ^< + & gt ^; +1)

Reference Example 1: Inhibitory activity of thrombin inhibitor

The enzyme inhibitory effect of the thrombin inhibitor, a compound according to the present invention, was determined by dissociation constant Ki determined by spectrophotometric method using a substrate that produces color upon reaction with the enzyme. That is, in the present invention, by using Chromozyme TH (tosyl-Gly-Pro-Arg-4-nitroanilide acetate) which is hydrolyzed by thrombin to produce yellow para-nitroanilide, The amount of nitroanilide is measured by changing the absorbance with time. From this rate, the activity of each enzyme can be measured and the ability of inhibitor to inhibit enzyme activity can be measured.

The ability of the compounds according to the present invention to inhibit thrombin activity was measured specifically by the methods described below.

To a 1.5 ml cuvette was added 1160 l 0.1 M Tris buffer solution (pH 7.8) containing 150 mM NaCl, 0.1% PEG 8000 (polyethyleneglycol, molecular weight about 8000). The substrate solution was prepared by dissolving crossover TH in dimethylsulfoxide (DMSO) at a concentration of 10 mM, and diluting it with the buffer solution to a concentration of 0.1 mM. 225 占 퐇 of the 0.1 mM substrate solution thus prepared was added to the cuvette. As the inhibitor solution, 10 mg / ml of the inhibitor compound according to the present invention was dissolved in dimethyl sulfoxide and diluted with the buffer solution to a concentration of 0.1 mg / ml, 0.01 mg / ml and 0.001 mg / ml, Was taken between 0 and 10 쨉 g, and then added to the cuvet with a total volume of 100 쨉 l using Tris buffer solution.

The cuvette containing the reaction solution at room temperature was added with 15 μl of thrombin dissolved in the above Tris buffer solution at a concentration of 0.1 mg / ml to start the enzymatic hydrolysis reaction. The amount of para-nitroanilide produced by the reaction for 2 minutes from the moment the enzyme was added was monitored by the change in absorbance at 381 nm to show a continuous spectrum of reaction time versus absorbance. Continuous spectra were obtained by performing the above experiments for various concentrations of inhibitor.

The inverse number (1 / Vi) graph of the initial velocity versus inhibitor concentration is shown after determining the initial velocity Vi from the slope within the initial 30 seconds of reaction time for each spectrum. After calculating the linear equation satisfying the points on the graph, we can calculate K i from the x-intercept of the equation using the following Michaelis-Menten equation. The Km value used in this calculation was obtained by varying the substrate concentration at a constant enzyme concentration of 8.3 μM.

The Michaelis-Menten equation

In the above enzyme reaction formula, [I] denotes the concentration of the inhibitor, [S] denotes the concentration of the substrate, Vmax denotes the maximum initial velocity, and Km denotes the Michaelis constant, which means 8.3 μM . The inhibitory potency of each inhibitor, measured against thrombin, as the Ki value is shown in Table 1 below.

The rate constant Ks is the same as that used for obtaining the Ki value, but the experimental method is different as described below.

That is, 1160 μl of a buffer solution was added to a 1.5 ml capacity cuvette, 15 μl of a 0.1 mg / ml thrombin solution and 100 μl of an inhibitor solution were added thereto, and the mixture was allowed to stand at room temperature for 15 minutes. Then 225 μl of a 0.1 mM substrate solution was added The change in absorbance with time was monitored for a minute. The slope of the portion showing a straight line in the obtained continuous spectrum is measured and expressed as Vs. This experiment was carried out at various inhibitor concentrations and a Vs value at each inhibitor concentration was determined to plot a 1 / Vs versus inhibitor concentration. After obtaining the first order equation satisfying the points on the graph, the Ks value was determined from the x-intercept of the graph using an enzyme reaction equation.

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

A 20 μM solution of N-benzoyl-valine-glycine-arginine para-nitroanilide hydrochloride (N-benzoyl-Val-Gly-Arg p-nitroanilide hydrochloride) was used as the substrate. Various concentrations were used. Also, trypsin was dissolved in 0.1 N HCl solution at a concentration of 45 μg / ml in the above Tris buffer solution immediately before the experiment, and 40 μl was used. Similar to the experiment on thrombin, the total volume of the reaction solution was 1.5 ml. In addition, the same method was used for the experiment. The K _M value used for the calculation of the Ki value was also determined to be 20.2 μM.

The ability of the inhibitor according to the present invention to inhibit the enzymatic activity of each of the thrombin and trypsin according to the above-described method is represented by Ki value and Ks value, and the selectivity to thrombin is represented by trypsin / thrombin. The results are shown in Table 1 below.

Inhibitor's ability to inhibit thrombin and trypsin The compound (Example No.) The ability to inhibit thrombin Ability to inhibit trypsin Selectivity (trypsin / thrombin) One Ks = 0.46 nM Ki = 3860 nM 8400 2 Ki = 0.53 nM 3 Ki = 0.66 nM 4 Ki = 2.37 nM 5 Ki = 0.60 nM 6 Ki = 1.40 nM 7 Ki = 1.40 nM 8 Ki = 1.71 nM 9 Ki = 1.46 nM 10 Ki = 3.5 nM 11 Ki = 0.28 nM Ki = 5000 nM 18,000 12 Ki = 3.7 nM 13 Ki = 38.5 nM 14 Ki = 0.30 nM Ki = 3150 nM 10,500 15 Ki = 2.7 nM

From the above experimental results, the compounds of the present invention exhibit excellent thrombin inhibitory activity, and in particular, the compounds of Examples 1, 11 and 14 exhibit the selectivities to trypsin versus thrombin at 8400, 18000 and 10500 times, respectively, Tropane) and 11 (NAPAP) were 250 times and 50 times, respectively.

Reference Example 2: Pharmacokinetic experiment

Experimental Method:

Male rats were fed after fasting for 24 hours. 1% (10 mg / ml) solution of the compound of Example 1 was prepared using physiological saline and then orally administered. Blood was collected at predetermined time intervals, and then methanol and zinc sulfate were immediately mixed. Then, the drug concentration in blood was measured at an ultraviolet wavelength of 231 nm using high pressure liquid chromatography (HPLC) on the upper layer solution.

Experiment result:

The measured results of blood drug concentration over time after oral administration of the compound of Example 1 are shown in Table 2 below. As can be seen from the results shown in Table 2, unlike the known thrombin inhibitor, agarose, the compound of Example 1 showed pharmacokinetic properties such that the drug was absorbed and retained in the blood for a long time in the blood upon oral administration.

When the compound of Example 1 was orally administered at a concentration of 30 mg / kg in rats, the blood drug concentration Time (minutes) Blood concentration (ng / ml) Average concentration (error) (ng / ml) Rat-1 Rat-2 Rat-3 Rat-4 5 168.93 161.45 114.83 109.29 138.62 (15.46) 10 216.11 233.54 313.01 253.18 253.96 (21.09) 20 452.22 552.59 435.03 425.81 466.41 (29.24) 40 444.70 958.28 813.32 636.75 713.26 (111.06) 60 436.39 1179.40 702.85 545.90 716.13 (163.82) 90 301.23 1506.94 539.15 535.90 720.81 (267.90) 120 234.01 1270.55 441.96 446.52 598.26 (229.51) 180 182.37 466.10 202.72 343.67 298.71 (66.33) 240 91.15 249.88 115.00 197.56 163.40 (36.75) 300 51.76 176.20 78.56 112.22 104.68 (26.86) 360 36.90 118.77 59.59 160.54 93.95 (28.12)

Claims (9)

Claims 1. A compound represented by the following formula (1): < EMI ID = [Chemical Formula 1] In this formula, R ¹ and R ² are each independently hydrogen, alkyl or alkyloxy, R ³ and R ⁴ each independently represent lower alkyl or cycloalkyl, R ⁵ represents hydrogen, lower alkyl or amino. 2. A compound according to claim 1, wherein R ¹ and R ² each independently represent hydrogen, C ₁ -C ₅ alkyl or C ₁ -C ₃ alkyloxy, R ³ and R ⁴ are each independently C ₁ -C ₃ alkyl or C ₃ -C ₅ cyclic alkyl, and R ⁵ represents C ₁ -C ₃ alkyl or amino. The compound according to claim 2, wherein R ¹ and R ² each independently represent hydrogen, methyl, ethyl, propyl, butyl or an isomer thereof, pentyl or an isomer thereof or ethyloxy, R ³ and R ⁴ are each independently methyl, ethyl Or cyclopentyl, and R < ⁵ > represents methyl or amino. 3. The compound of claim 1 which is (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N- (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-butyl-benzenesulfonylamino) ) - N-cyclopentyl-N-methyl-2- (4-t-butyl-benzenesulfonylamino) - propionamide, Methyl-2- (3,4-diethyl-benzenesulfonylamino) propionamide, (S) -3- (3-methyl-4-propyl-benzenesulfonylamino) propionamide, (S) -3- (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-ethyl- benzenesulfonylamino) propionamide, (S ) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N- 2- (3-methyl-4-ethyl-benzenesulfonylamino) propionamide, (S) -3- (4-amidazo no- Phenyl) -N-cyclopentyl-N-methyl-2- (4-propyl-isobutyl-benzenesulfonylamino) (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-2- (4-isobutyl-benzenesulfonylamino) -N-methyl- propionamide, (S) -3- [4- (amino-methylimino-methyl) -phenyl] -N-cyclopentyl-2- (4-isobutyl-benzenesulfonylamino) Methyl-propionamide, (S) -3- (4-amide-2-methyl- Methyl-propionamide, (S) -3- [4- (amino < RTI ID = 0.0 > -Methylimino-methyl) -phenyl] -N-cyclopentyl-2- (4- (2,2-dimethoxy- (S) -3- (4-amidorazono-phenyl) -N-cyclopentyl-N-methyl-2- (4-ethyloxy) -benzenesulfonylamino] -Benzenesulfonylamino) -propionamide. ≪ / RTI > And reacting the methyl mercapto compound represented by the following formula (2) with an amine derivative represented by the following formula (3): < EMI ID = [Chemical Formula 1] (2) (3) In this formula, R ¹ and R ² are each independently hydrogen, alkyl or alkyloxy, R ³ and R ⁴ each independently represent lower alkyl or cycloalkyl, R ⁵ represents hydrogen, lower alkyl or amino. 6. The process according to claim 5, wherein the reaction is carried out in the presence of a solvent. 7. The method of claim 6, wherein the solvent is an alcohol. A thrombin inhibitor composition comprising a compound of formula (I) according to claim 1 as an active ingredient together with a pharmaceutically acceptable carrier. The composition according to claim 8, which is formulated into an oral dosage form.