KR100227729B1 - Novel butylamine-based hydantoin derivatives useful as thrombin inhibitor - Google Patents

Abstract

The present invention contains a novel butylamine-based hydridein derivative of the general formula (1), a pharmaceutically acceptable salt and an isomer thereof, a preparation method thereof, and a compound of the general formula (1), which are easily synthesized and exhibit excellent thrombin inhibitory activity. It relates to a pharmaceutical composition for preventing blood clotting or treating various thrombosis.

In the above formula,

R ¹ represents alkyl, cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl at the aryl moiety, or substituted or unsubstituted heterocyclylalkyl,

R ² represents alkyl, aralkyl unsubstituted or substituted at the aryl moiety, or in each case substituted or unsubstituted aryl or heterocyclyl.

Description

Novel Butylamine-based Hydientin Derivatives Useful as Thrombin Inhibitors

The present invention relates to nonpeptidic hydrentin derivatives useful as thrombin inhibitors. More specifically, the present invention is a novel butylamine-based hydridein derivative of Formula 1 below, which is easy to synthesize and exhibits strong thrombin inhibitory activity, a preparation method thereof, and a blood clotting prevention or various thrombosis treatment containing the compound as an active ingredient. It relates to a pharmaceutical composition for.

[Formula 1]

In the above formula,

R ¹ represents alkyl, cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl at the aryl moiety, or substituted or unsubstituted heterocyclylalkyl,

R ² represents alkyl, aralkyl unsubstituted or substituted at the aryl moiety, or in each case substituted or unsubstituted aryl or heterocyclylalkyl.

The blood clotting mechanism involves a number of complex enzymatic reactions, the final stage of which involves the conversion of prothrombin to thrombin, which activates platelets and converts fibrinogens to fibrin. It plays an important role, in which fibrin is converted into a polymer by a polymerization reaction and crosslinked by activated blood factor XIII to become an insoluble blood clot. Thrombin also activates blood factors V and VIII, which accelerate blood coagulation by feedback. Thus, inhibitors of thrombin may act as effective anticoagulants by inhibiting the activation of platelets and preventing the production of fibrin.

Peptides with cleavage sites similar to fibrin, the substrate of thrombin, are three amino acids, D-Phe-Pro-Arg, and derivatives mimicking this structure have been reported to be effective as thrombin inhibitors. These derivatives are compounds in which the C-terminal portion of D-Phe-Pro-Arg is substituted with aldehyde, phosphonate or boronic acid (Trends in Pharm. Sci. 1993, 14, 366-376). However, since most of these compounds retain the properties of peptides, peptide-based compounds are generally unstable in vivo, cannot be absorbed into organs, and have fast degrading properties. Not suitable Therefore, it is very difficult to develop a peptide-based compound as a useful thrombin agent.

Therefore, various studies have recently been made to develop nonpeptidic compounds as thrombin inhibitors. Representative non-peptidic thrombin inhibitors are agatroban represented by the following formula (2) commercialized in Japan in 1990 (see US Pat. Nos. 94258192 and 4201863).

Agatroban, however, is an arylsulfonyl arginine-based compound, which has excellent efficacy as an injection, but is not active at all for oral administration, and has a disadvantage in that it is prepared only through a complicated synthesis process.

Therefore, the present inventors have conducted extensive research to develop a non-peptide thrombin inhibitor compound having a novel structure that can be easily synthesized and exhibit effective thrombin inhibitory activity, and as a result, the compound of formula 1 as defined above is It was confirmed that can be achieved and completed the present invention.

Accordingly, it is an object of the present invention to provide a butylamine-based hydridein derivative represented by the formula (1), which is easy to synthesize and exhibits useful thrombin inhibitory activity, and pharmaceutically acceptable salts and isomers thereof.

[Formula 1]

In the above formula,

R ¹ represents alkyl, cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl at the aryl moiety, or substituted or unsubstituted heterocyclylalkyl,

R ² represents alkyl, aralkyl unsubstituted or substituted at the aryl moiety, or in each case substituted or unsubstituted aryl or heterocyclylalkyl.

Still another object of the present invention is to provide a method for preparing the compound of Formula 1 and a pharmaceutical composition for preventing blood clotting or treating various thrombosis containing the compound as an active ingredient.

As used herein, the term "alkyl" as used to define a substituent of a compound of Formula 1 means straight or branched chain alkyl of 1 to 8 carbon atoms including methyl, ethyl, isopropyl, isobutyl, t-butyl, octyl, and the like. The term "lower alkyl" refers to a straight or branched chain alkyl having 1 to 4 carbon atoms among these alkyl groups, and the term "cycloalkyl" means a cyclic alkyl group having 3 to 8 carbon atoms including cyclohexyl.

The term "aryl" in the terms "aryl" and "aralkyl" means a 6- to 14-membered monocyclic to tricyclic aromatic group, which groups are unsubstituted or lower alkyl, lower alkoxy, aryl and aryloxy It may be mono- or polysubstituted by a substituent selected from the group consisting of: The term "alkyl" in the term "aralkyl" denotes the meaning as mentioned above.

In the terms "heterocyclyl" and "heterocyclylalkyl" "heterocyclyl" means a heterocyclic group condensed with a carbocyclic ring having a ring containing one or two oxygen atoms as ring member, and "lower" Alkyl "means an alkyl group as defined above.

Preferred among the compounds of the formula (1) according to the present invention are phenyl, benzyl or benzo wherein R ¹ is unsubstituted or substituted by C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₄ alkoxy , 3] dioxol-5-ylmethyl, R ² is C ₁ -C ₄ alkyl, aryl moiety selected from the group consisting of C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, phenyl and benzyloxy Aryl or aryl-C ₁ -C ₄ alkyl which is a 6- to 14-membered monocyclic to tricyclic aromatic group unsubstituted or mono- to trisubstituted, or unsubstituted by C ₁ -C ₄ lower alkoxy Or mono- or di-substituted 2,3-dihydro-benzo [1,4] dioxyyl or tetrahydrofuranyl.

Particularly preferred among the compounds of formula 1 according to the invention are those wherein R ¹ represents ethyl, cyclohexyl, benzyl, phenyl, ethoxyphenyl or benzo [1,3] dioxol-5-ylmethyl, R ² represents propyl, Phenyl, naphthyl, methoxyphenyl, benzyloxyphenyl, biphenyl, diphenylmethyl, dimethoxymethylphenyl, anthracenyl, bisbenzyloxyphenyl, 2,3-dihydro-benzo [1,4] dioxyyl or 2 , 5-dimethoxytetrahydrofuranyl.

As specific examples of the compound of formula 1 according to the present invention, the following compounds may be mentioned.

1) (S) -5- (4-amino-butyl) -1- (naphthalen-2-ylmethyl) -3-phenyl-imidazolidine-2,4-dione,

2) (S) -5- (4-amino-butyl) -3-ethyl-1- (naphthalen-1-ylmethyl) -imidazolidine-2,4-dione,

3) (S) -5- (4-amino-butyl) -3-benzyl-1- (naphthalen-1-ylmethyl) -imidazolidine-2,4-dione,

4) (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1-benzyl-imidazolidine-2,4-dione,

5) (S) -5- (4-Amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (4-methoxy-benzyl) -imidazolidine- 2,4-dione,

6) (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (2,3-dihydro-benzo [1,4] Dioxin-6-ylmethyl) -imidazolidine-2,4-dione,

7) (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (naphthalen-2-ylmethyl) -imidazolidine- 2,4-dione,

8) (S) -5- (4-Amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (4-benzyloxy-benzyl) -imidazolidine- 2,4-dione,

9) (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (biphenyl-4-ylmethyl) -imidazolidine -2,4-dione,

10) (S) -5- (4-amino-butyl) -3-ethyl-1-isobutyl-imidazolidine-2,4-dione,

11) (S) -5- (4-amino-butyl) -1-isobutyl-3-phenyl-imidazolidine-2,4-dione,

12) (S) -5- (4-amino-butyl) -3-cyclohexyl-1-isobutyl-imidazolidine-2,4-dione,

13) (S) -5- (4-amino-butyl) -3-benzyl-1-isobutyl-imidazolidine-2,4-dione,

14) (S) -5- (4-amino-butyl) -3- (4-ethoxy-phenyl) -1-isobutyl-imidazolidine-2,4-dione,

15) (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1-isobutyl-imidazolidine-2,4-dione,

16) (S) -5- (4-amino-butyl) -1- (2,4-dimethoxy-3-methyl-benzyl) -3-ethyl-imidazolidine-2,4-dione,

17) (S) -5- (4-amino-butyl) -1- (2,5-dimethoxy-tetrahydrofuran-3-ylmethyl)-3-ethyl-imidazolidine-2,4-dione ,

18) (S) -5- (4-amino-butyl) -1- (2,2-diphenyl-ethyl) -3-ethyl-imidazolidine-2,4-dione,

19) (S) -5- (4-amino-butyl) -1- (anthracene-9-ylmethyl) -3-ethyl-imidazolidine-2,4-dione,

20) (S) -5- (4-amino-butyl) -1- (3,5-bis-benzyloxy-benzyl) -3-ethyl-imidazolidine-2,4-dione,

21) (S) -5- (4-amino-butyl) -3-ethyl-1- (naphthalen-2-ylmethyl) -imidazolidine-2,4-dione,

22) (S) -5- (4-amino-butyl) -1- (naphthalen-2-ylmethyl) -3-phenyl-imidazolidine-2,4-dione,

23) (S) -5- (4-amino-butyl) -3-benzyl-1- (naphthalen-2-ylmethyl) -imidazolidine-2,4-dione,

24) (S) -5- (4-amino-butyl) -3- (4-ethoxy-phenyl) -1- (4-methoxy-benzyl) -imidazolidine-2,4-dione.

The structures of the most representative compounds among these compounds according to the present invention are shown in Table 1 below.

Compound number Compound structure Compound number Compound structure One

4

2

5

3 6

Compound number Compound structure Compound number Compound structure 7

22

9

23

21

24

The compounds of formula 1 according to the invention may also form pharmaceutically acceptable salts. Such pharmaceutically acceptable salts include acids that form non-toxic acid addition salts containing pharmaceutically acceptable anions such as inorganic acids, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, tartaric acid, formic acid, Organic carbonic acid such as citric acid, 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 Acid addition salts formed by and the like are included.

On the other hand, the compounds according to the invention may have an asymmetric carbon center and therefore may exist as racemic compounds, diastereomeric mixtures and individual diastereomers, all of these isomers being included within the scope of the invention.

The invention also relates to a process for preparing the compound of formula 1 as defined above.

According to the present invention, the compound of formula 1 is synthesized using combinatorial chemistry. Combinatorial Synthesis uses a solid phase, which uses a new microtechnology that is completely different from the conventional methods of pharmaceutical synthesis, so that many different compounds can be produced simultaneously in a short time and at a low cost. Many companies have been trying new ways (see HM Geysen et al., Proc. Natl. Acad. Sci, USA, 81, 3998, 1984; MA Gallop et al., J. Med. Chem., 37, 1233, 1994; EM Gordon et al., J. Med. Chem. 37, 1386, 1994; JA Ellman et al., Chem. Rev. 1996, 96, 555-600). In the case of synthesizing a target compound using a combinatorial chemistry using a solid phase, all compounds can be prepared in various ways by the same manufacturing method.

According to the present invention, the target compound of formula 1 may be prepared by reacting a resin-bonded carbamate compound represented by formula 3 with diisopropylamine to form a compound of formula 4, and then removing the amino protecting group. have.

In the above formula,

R ¹ and R ² are as defined above,

P represents an amino-protecting group.

In general ring-closure reaction, unlike the reaction using a solvent and a strong acid or base, in the ring-closure reaction of the method according to the present invention, only the diisopropylamine is used to prepare the target compound. It also has the advantage that it does not need to use a separate solvent. This method is the first attempt in the field of pharmaceutical manufacture and has the advantage of being able to produce the desired product quantitatively, i.e. in a yield of almost 100%. Method for preparing a compound of formula 1 according to the present invention can be represented by the following scheme 1.

Wherein R ¹ , R ² and P are each as defined above.

The method of the present invention will be described in detail as follows. First, the desired compound of formula 1 can be obtained by a simple one step process by treating the resin-bonded carbamate compound of formula 3 with diisopropylamine and then performing a conventional amino-protecting group removal process. have. Wang resin, which is bound to the compound of Formula 3, has excellent properties as a leaving group, and is manufactured and marketed by various companies (eg, Advanced ChemTech, Novabiochem, etc.). In this reaction, triethylamine, pyrrolidine, piperidine, and the like were used instead of diisopropylamine, but similar effects were found. However, in order to obtain a target compound, the solvent used was easily removed. It is preferable to use diisopropylamine which should be able to be present and has a low boiling point in this respect.

After treating the compound of formula 3 with diisopropylamine to separate the resin, the desired compound of formula 1 is obtained by removing the amino protecting group by conventional methods. In the present invention, particularly preferably, the amino protecting group can be removed by treating the intermediate separated by treatment with diisopropylamine with trifluoroacetic acid in a solvent.

In general, products by combinatorial chemistry can yield the desired product in almost 100% yield without the need for additional post-treatment methods such as chromatography, recrystallization and the like.

The resin-bonded carbamate compound of formula 3 used as starting material in the method shown in Scheme 1 is condensed with the resin represented by formula 6 by condensation of the L-lysine derivative of formula 5 with a condensing agent and a base. To obtain a compound of formula 8 by removing the 9-proenylmethoxycarbonyl (FMOC) protecting group from the compound of formula 7, and then reacting the compound of formula 8 with an aldehyde compound of formula 9 Can be prepared by reacting the compound of Formula 10 with the isocyanate compound of Formula 11.

Wherein R ¹ , R ² and P are as defined above.

Method for preparing a compound of formula 3 as described above can be represented by the following scheme 2.

The preparation method of the compound of formula 3 shown in Scheme 2 will be described together with specific reaction conditions thereof. That is, L-lysine with N-α-9-proenylmethoxycarbonyl (FMOC) -N-ε-butoxycarbonyl (BOC) protecting group, that is, 3.5 equivalents of the compound of formula 5 is equivalent to 1 equivalent of the resin of formula 6 With diisopropylcarbodiimide (DIC) 3.5 equivalents, 4-dimethylaminopyridine (DMAP) in the presence of 0.1 equivalents in N, N-dimethylformamide (DMF) solvent for one day and then excess diisopropyl Carbodiimide (DIC), lysine, and DMF solvent are filtered off to obtain a resin-bound compound of formula (7). After the reaction is completed, the loading level of the resin is about 0.81 mmol / g. The compound of formula 7 is then stirred for 1 hour in the presence of 20% piperidine solution and DMF solvent to remove the existing 9-proenylmethoxycarbonyl (FMOC) protecting group to afford the compound of formula 8. Kaiser test is carried out to confirm the completion of the protecting group removal reaction. The resin turns dark blue when the 9-proenylmethoxycarbonyl (FMOC) protecting group is completely removed. In this way, the compound of formula (8) identified as having removed the protecting group is reacted with an aldehyde compound of formula (9) to obtain a compound of formula (10) by reductive amination, which is then reacted with an isocyanate compound of formula (11). This can yield the compound of formula 3, which is the desired intermediate.

As mentioned above, the compound of formula 1 according to the present invention has a selective inhibitory effect on thrombin, and thus is useful for preventing blood coagulation and for treating thrombosis.

Accordingly, another object of the present invention is to provide a pharmaceutical composition for preventing blood clotting and treating thrombosis, which contains the compound of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

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 0.1 mg to 10 mg per kg of body weight, but the specific dose level for a particular patient may be used. Specific compounds, body weight, sex, health status, diet, time of administration, method of administration, rate of excretion, drug mixture and severity of disease may be appropriately determined.

The compounds of the present invention may preferably be formulated in injectable preparations.

Injectable preparations, such as sterile injectable aqueous or oleaginous suspensions, can be prepared using suitable dispersing agents, wetting agents, or suspending agents according to known techniques. Pharmaceutically 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-, diglycerides, and also fatty acids such as oleic acid are used in the preparation of injectables.

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 activity inhibitors include aspirin, ticlopidin, Clopidrogel, 7E3 monoclonal antibody, 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 examples and experimental examples, but the present invention is not limited in any way by these examples.

Example 1: (S) -5- (4-amino-butyl) -1- (naphthalen-2-ylmethyl) -3-phenyl-imidazolidine-2,4-dione

Into a glass reaction vessel equipped with Teflon frit (30 mg), 30 mg of a butylamine compound of formula (8) having an aqua resin bound was added, followed by 19.5 mg of 2-naphthalenecarboaldehyde, 1.0 ml of N, N-dimethylacetamide solvent, and 1 0.1 ml of% acetic acid was added and stirred at 25 ° C. for 5 hours. Then, 7.8 mg of sodium cyanoborohydride (NaBH ₃ CN) was added and stirred for one day, followed by suction filtration to remove excess solvent and naphthalene carboaldehyde. The residue was washed three times with 10 ml of methylene clyde and 10 ml of methanol, followed by Kaiser test to confirm that the reaction was completed. 14.8 mg of phenyl isocyanate and dimethyl were added to the product in the reaction vessel. 1.0 ml of a mixed solvent of formamide and toluene was added, the mixture was stirred for 5 hours, and then filtered again. After completely drying the product, 1.0 ml of diisopropylamine was added and stirred for 5 hours, followed by filtration. The filtrate was completely dried, and then 1 ml of trifluoroacetic acid and 1 ml of methylene chloride were added to react for 30 minutes, followed by drying. 6.7 mg (total yield: 70%) of the title compound were obtained.

¹ HNMR (CDCl ₃ , ppm); δ 8.21 (d, 1H, J = 10.0 Hz), 7.90 (m, 2H), 7.60-7.30 (m, 9H), 5.55 (d, 1H, J = 7.9 Hz), 4.71 (d, 1H, J = 14.0 Hz), 4.48 (s, 1H), 2.99 (m, 2H), 1.93 (m, 2H), 1.30 (m, 4H)

Mass (FAB, m / e); 388 (M ⁺ +1)

Example 2: (S) -5- (4-amino-butyl) -3-ethyl-1- (naphthalen-1-ylmethyl) -imidazolidine-2,4-dione

The reaction was carried out according to the same method as in Example 1, except that 19.5 mg of aldehyde 1-naphthalenecarboaldehyde and 9.0 mg of ethyl isocyanate were used as the isocyanate, and the title compound was 5.7 mg (total yield: 68% ) Was obtained.

¹ HNMR (CDCl ₃ , ppm); δ8.14 (d, 1H, J = 10.0 Hz), 7.85 (m, 4H), 7.51 (m, 2H), 5.45 (d, 1H, J = 7.9 Hz), 4.61 (d, 1H, J = 15.0 Hz ), 4.38 (s, 1H), 3.60 (m, 2H), 3.00 (m, 2H), 1.80 (m, 2H), 1.27 (m, 6H), 0.9 (t, 3H, J = 3.7 Hz)

Mass (FAB, m / e): 340 (M ⁺ +1)

Example 3: (S) -5- (4-amino-butyl) -3-benzyl-1- (naphthalen-1-ylmethyl) -imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 19.5 mg of aldehyde 1-naphthalenecarboaldehyde and 16.6 mg of benzyl isocyanate were used as the isocyanate, to give 6.5 mg of the title compound (total yield: 65% ) Was obtained.

¹ HNMR (CDCl ₃ , ppm); δ 8.18 (d, 1H, J = 9.9 Hz), 7.85 (m, 2H), 7.40-6.90 (m, 9H), 5.57 (d, 1H, J = 7.9 Hz), 4.47 (m, 4H), 2.99 (m, 2H), 1.90 (m, 2H), 1.32 (m, 4H)

Mass (FAB, m / e); 402 (M ⁺ +1)

Example 4 (S) -5- (4-Amino-butyl) -3- (benzo [1,3] -dioxol-5-ylmethyl) -1-benzyl-imidazolidine-2,4- Dion

The reaction was carried out according to the same method as in Example 1, except that 13.2 mg of aldehyde benzaldehyde and 22.1 mg of piperonyl isocyanate were used as the isocyanate, to obtain 6.9 mg (yield: 70%) of the title compound. .

Mass (FAB, m / e): 396 (M ⁺ +1)

Example 5 (S) -5- (4-Amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (4-methoxy-benzyl) -imidazoli Dean-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 17.0 mg of aldehyde 4-methoxybenzaldehyde was used and 16.6 mg of piperonyl isocyanate as the isocyanate, yielding 7.3 mg of the title compound (yield: 69% ) Was obtained.

Mass (FAB, m / e): 426 (M ⁺ +1)

Example 6: (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (2,3-dihydro-benzo [1, 4] dioxine-6-ylmethyl) -imidazolidine-2,4-dione

Same as in Example 1 except using 20.5 mg of aldehyde 2,3-dihydro-benzo [1,4] dioxin-6-carboaldehyde and 16.6 mg of piperonyl isocyanate as the isocyanate The reaction was carried out according to the method, obtaining 8.0 mg (yield: 71%) of the title compound.

Mass (FAB, m / e): 454 (M ⁺ +1)

Example 7: (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (naphthalen-2-ylmethyl) -imidazoli Dean-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 19.5 mg of aldehyde 2-naphthalenecarboaldehyde was used and 16.6 mg of piperonyl isocyanate as the isocyanate, yielding 7.1 mg of the title compound (yield: 64 %) Was obtained.

Mass (FAB, m / e): 446 (M ⁺ +1)

Example 8: (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (4-benzyloxy-benzyl) -imidazoli Dean-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 26.5 mg of aldehyde 4-benzyloxybenzaldehyde and 16.6 mg of piperonyl isocyanate were used as the isocyanate, and the title compound was 7.5 mg (yield: 60%). ) Was obtained.

Mass (FAB, m / e): 502 (M ⁺ +1)

Example 9: (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (biphenyl-4-ylmethyl) -imida Zolidine-2,4-dione

The reaction was carried out according to the same method as in Example 1, using 22.7 mg of aldehyde 4-phenyl-benzaldehyde and 22.1 mg of piperonyl isocyanate as the isocyanate (yield: 58%) ) Was obtained.

Mass (FAB, m / e): 472 (M ⁺ +1)

Example 10 (S) -5- (4-Amino-butyl) -3-ethyl-1-isobutyl-imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1 except that 9.0 mg of aldehyde isobutylaldehyde was used and 9.0 mg of ethyl isocyanate as the isocyanate to obtain 4.0 mg (yield: 63%) of the title compound. .

Mass (FAB, m / e): 256 (M ⁺ +1)

Example 11 (S) -5- (4-Amino-butyl) -1-isobutyl-3-phenyl-imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 9.0 mg of aldehyde isobutylaldehyde was used and 14.9 mg of phenyl isocyanate as the isocyanate, to obtain 5.1 mg (yield: 67%) of the title compound. .

Mass (FAB, m / e): 304 (M ⁺ +1)

Example 12 (S) -5- (4-Amino-butyl) -3-cyclohexyl-1-isobutyl-imidazolidine-2,4-dione

The reaction was carried out according to the same method as in Example 1, except that 9.0 mg of aldehyde isobutylaldehyde was used and 15.6 mg of cyclohexyl isocyanate as the isocyanate, to obtain 4.9 mg (yield: 64%) of the title compound. It was.

Mass (FAB, m / e): 310 (M ⁺ +1)

Example 13 (S) -5- (4-Amino-butyl) -3-benzyl-1-isobutyl-imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 9.0 mg of aldehyde isobutylaldehyde was used and 15.6 mg of benzyl isocyanate as the isocyanate, to obtain 4.8 mg (yield: 60%) of the title compound. .

Mass (FAB, m / e): 318 (M ⁺ +1)

Example 14 (S) -5- (4-Amino-butyl) -3- (4-ethoxy-phenyl) -1-isobutyl-imidazolidine-2,4-dione

The reaction was carried out according to the same method as in Example 1, except that 9.0 mg of aldehyde isobutylaldehyde was used and 20.3 mg of 4-ethoxyphenyl isocyanate as the isocyanate (Yield: 66%) ) Was obtained.

Mass (FAB, m / e): 348 (M ⁺ +1)

Example 15 (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1-isobutyl-imidazolidine-2,4- Dion

The reaction was carried out according to the same method as in Example 1, except that 9.0 mg of aldehyde isobutylaldehyde and 22.1 mg of piperonyl isocyanate were used as the isocyanate to give 5.2 mg (yield: 58%) of the title compound. Obtained.

Mass (FAB, m / e): 362 (M ⁺ +1)

Example 16 (S) -5- (4-Amino-butyl) -1- (2,4-dimethoxy-3-methyl-benzyl) -3-ethyl-imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 22.5 mg of aldehyde 2,4-dimethoxy-3-methyl-benzaldehyde was used and 9.0 mg of ethyl isocyanate as the isocyanate to give 5.5 mg of the title compound. (Yield 61%) was obtained.

Mass (FAB, m / e): 364 (M ⁺ +1)

Example 17 (S) -5- (4-Amino-butyl) -1- (2,5-dimethoxy-tetrahydrofuran-3-ylmethyl) -3-ethyl-imidazolidine-2,4 Dion

The reaction was carried out in the same manner as in Example 1 except that 20.0 mg of aldehyde 2,5-dimethoxy-tetrahydrofuran-3-carboaldehyde and 9.0 mg of ethyl isocyanate were used as the isocyanate 6.0 mg (yield 57%) of the title compound were obtained.

Mass (FAB, m / e): 425 (M ⁺ +1)

Example 18: (S) -5- (4-amino-butyl) -1- (2,2-diphenyl-ethyl) -3-ethyl-imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 24.5 mg of aldehyde 2,2-diphenylacetaldehyde was used and 9.0 mg of ethyl isocyanate as the isocyanate, yielding 3.8 mg of the title compound (yield: 61 %) Was obtained.

Mass (FAB, m / e): 380 (M ⁺ +1)

Example 19: (S) -5- (4-amino-butyl) -1- (anthracene-9-ylmethyl) -3-ethyl-imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 25.7 mg of aldehyde anthracenecarboaldehyde and 9.0 mg of ethyl isocyanate were used as the isocyanate to obtain 5.8 mg (yield: 60%) of the title compound. It was.

Mass (FAB, m / e): 390 (M ⁺ +1)

Example 20 (S) -5- (4-Amino-butyl) -1- (3,5-bis-benzyloxy-benzyl) -3-ethyl-imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 39.7 mg of aldehyde 3,5-bis-benzyloxy-benzaldehyde was used and 9.0 mg of ethyl isocyanate as the isocyanate, yielding 6.7 mg of the title compound (yield: : 55%) was obtained.

Mass (FAB, m / e): 490 (M ⁺ +1)

Example 21 (S) -5- (4-amino-butyl) -3-ethyl-1- (naphthalen-2-ylmethyl) -imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 19.5 mg of aldehyde 2-naphthalenecarboaldehyde and 9.0 mg of ethyl isocyanate were used as the isocyanate, and the title compound was 4.9 mg (yield: 59%). Obtained.

Mass (FAB, m / e): 338 (M ⁺ +1)

Example 22: (S) -5- (4-amino-butyl) -1- (naphthalen-2-ylmethyl) -3-phenyl-imidazolidine-2,4-dione

The reaction was carried out according to the same method as in Example 1, except that 19.5 mg of aldehyde 2-naphthalenecarboaldehyde and 20.3 mg of phenyl isocyanate were used as the isocyanate, and 6.1 mg (yield: 63%) of the title compound. Obtained.

Mass (FAB, m / e): 386 (M ⁺ +1)

Example 23 (S) -5- (4-amino-butyl) -3-benzyl-1- (naphthalen-2-ylmethyl) -imidazolidine-2,4-dione

The reaction was carried out according to the same method as in Example 1, except that 19.5 mg of aldehyde 2-naphthalenecarboaldehyde and 16.6 mg of benzyl isocyanate were used as the isocyanate, to obtain 6.3 mg of the title compound (yield: 63%). Obtained.

Mass (FAB, m / e): 400 (M ⁺ +1)

Example 24 (S) -5- (4-Amino-butyl) -3- (4-ethoxy-phenyl) -1- (4-methoxy-benzyl) -imidazolidine-2,4-dione

The reaction was carried out in the same manner as in Example 1, except that 17.0 mg of aldehyde 4-methoxy-benzaldehyde was used and 20.3 mg of 4-ethoxyphenyl isocyanate as the isocyanate, yielding 6.7 mg of the title compound (yield: : 65%) was obtained.

Mass (FAB, m / e): 412 (M ⁺ +1)

Experimental Example: Inhibitory Activity of a Thrombin Inhibitor

The inhibitory effect on thrombin of the compound of formula 1 according to the present invention was determined by the rate constant Ki measured spectrophotometrically using a substrate which reacts with an enzyme to produce color. The rate constant Ki represents the extent to which the inhibitor inhibits the enzyme activity when the enzyme, substrate and inhibitor are added simultaneously. Hydrolysis of thrombin-based substrate, Chromozym TH (Tosyl-Gly-Arg-4-nitroanilide acetate), produces para-nitroaniline. The amount of yellow para-nitroaniline thus produced was measured as a change in absorbance over time. From this rate, the activity of the enzyme can be measured, and the inhibitory ability of the inhibitor can be measured.

The inhibitory ability of the inhibitor against thrombin was determined by the following method.

To a 1.5 ml cuvette, add 1160 μl of 0.1 M Tris buffer solution (pH 7.8) containing 150 mM NaCl and 0.1% PEG8000 (polyethylene glycol, molecular weight about 8,000), and then add a pigment-forming substrate, Chromozyme TH 0.1 mM solution 225 Μl was added. Chromozyme TH was dissolved in dimethylsulfoxide (DMSO) so as to have a concentration of 10 mM, and diluted with the buffer solution at a concentration of 0.1 mM. The thrombin inhibitor of the present invention was dissolved in dimethylsulfoxide to a concentration of 10 mg / ml, and then diluted with Tris buffer solution having the composition as mentioned above to make 0.1 mg / ml and 0.001 mg / ml. After the solution was taken in the range of 0 to 10 µg, the Tris buffer solution was added to the final volume of 100 µl and added to each cuvette.

15 µl of a solution of sothrombin dissolved in the Tris buffer solution at a concentration of 0.1 mg / ml was added to the cuvette containing the reaction solution at room temperature to start the enzymatic hydrolysis reaction. The amount of para-nitroaniline produced by the reaction for 2 minutes from the moment of addition of the enzyme was monitored by a change in absorbance at 381 nm to draw a continuous spectrum of reaction time versus absorbance. The experiment was repeated at different inhibitor concentrations to obtain continuous spectra.

After obtaining the initial velocity Vi from the slope within 30 seconds of the initial reaction time in each reaction, a graph of the inverse of the initial velocity versus the inhibitor concentration (1 / Vi) is shown. After calculating the first equation satisfying the points on the graph, Ki was calculated from the x-intercept of the equation using the enzyme reaction equation. The K _M value used in this calculation was 8.3 μM, which was obtained by varying the substrate concentration at a constant enzyme concentration.

Michaelis-Menten equation

In the enzyme scheme, [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 is 8.3 μM. .

Table 2 shows the enzyme activity inhibitory capacity of each inhibitor measured for thrombin as Ki value.

Inhibitory activity (Ki value) of thrombin of the inhibitor according to the present invention Inhibitor Compound (Example Number) Ki value (μM) Inhibitor Compound (Example Number) Ki value (μM) One 20 19 15.9 4 100 21 29 9 25.2 22 28.3 16 15.3 23 40 18 15.4 24 64.2

From the above experimental results, it can be seen that the compound of the present invention has superior selectivity to thrombin relative to trypsin, compared with known compounds.

Claims (6)

Compound represented by the following formula (1), pharmaceutically acceptable salts and isomers thereof: [Formula 1]

In the above formula, R ¹ represents alkyl, cycloalkyl, substituted or unsubstituted aryl, heterocyclylalkyl condensed with a carbocycle having a ring containing one or two oxygen atoms substituted or unsubstituted at the aryl moiety, R ² has a straight or branched chain alkyl of 1 to 8 carbon atoms, an aralkyl substituted or unsubstituted at an aryl moiety, or in each case a substituted or unsubstituted aryl or a ring containing one or two oxygen atoms and Condensed heterocyclyl is shown. The phenyl, benzyl or benzo [1,3] dioxol- of claim 1, wherein R ¹ is unsubstituted or substituted by C ₁ -C ₄ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₄ alkoxy. 5-ylmethyl, R ² is C ₁ -C ₄ alkyl, the aryl moiety is unsubstituted or substituted by a substituent selected from the group consisting of C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, phenyl and benzyloxy Unsubstituted or mono-disubstituted by aryl or aryl-C ₁ -C ₄ alkyl, or C ₁ -C ₄ lower alkoxy, which is a substituted to trisubstituted 6- to 14-membered monocyclic to tricyclic aromatic group 2,3-dihydro-benzo [1,4] dioxyyl or tetrahydrofuranyl. The compound of claim 2, wherein R ¹ represents ethyl, cyclohexyl, benzyl, phenyl, ethoxyphenyl or benzo [1,3] dioxol-5-ylmethyl, and R ² represents propyl, phenyl, naphthyl, methoxy Phenyl, benzyloxyphenyl, biphenyl, diphenylmethyl, dimethoxymethylphenyl, anthracenyl, bisbenzyloxyphenyl, 2,3-dihydro-benzo [1,4] dioxyyl or 2,5-dimethoxytetrahydro A compound representing furanyl. The compound according to claim 1, wherein (S) -5- (4-amino-butyl) -1- (naphthalen-2-ylmethyl) -3-phenyl-imidazolidine-2,4-dione, (S)- 5- (4-Amino-butyl) -3-ethyl-1- (naphthalen-1-ylmethyl) -imidazolidine-2,4-dione, (S) -5- (4-amino-butyl)- 3-benzyl-1- (naphthalen-1-ylmethyl) -imidazolidine-2,4-dione, (S) -5- (4-amino-butyl) -3- (benzo [1,3] di Oxol-5-ylmethyl) -1-benzyl-imidazolidine-2,4-dione, (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5 -Ylmethyl) -1- (4-methoxy-benzyl) -imidazolidine-2,4-dione, (S) -5- (4-amino-butyl) -3- (benzo [1,3] Dioxol-5-ylmethyl) -1- (2,3-dihydro-benzo [1,4] dioxine-6-ylmethyl) -imidazolidine-2,4-dione, (S) -5 -(4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (naphthalen-2-ylmethyl) -imidazolidine-2,4-dione, ( S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (4-benzyloxy-benzyl) -imidazolidine-2,4 -Dione, (S) -5- (4-amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1- (ratio Phenyl-4-ylmethyl) -imidazolidine-2,4-dione, (S) -5- (4-amino-butyl) -3-ethyl-1-isobutyl-imidazolidine-2,4 -Dione, (S) -5- (4-amino-butyl) -1-isobutyl-3-phenyl-imidazolidine-2,4-dione, (S) -5- (4-amino-butyl) -3-cyclohexyl-1-isobutyl-imidazolidine-2,4-dione, (S) -5- (4-amino-butyl) -3-benzyl-1-isobutyl-imidazolidine- 2,4-dione, (S) -5- (4-amino-butyl) -3- (4-ethoxy-phenyl) -1-isobutyl-imidazolidine-2,4-dione, (S) -5- (4-Amino-butyl) -3- (benzo [1,3] dioxol-5-ylmethyl) -1-isobutyl-imidazolidine-2,4-dione, (S) -5 -(4-Amino-butyl) -1- (2,4-dimethoxy-3-methyl-benzyl) -3-ethyl-imidazolidine-2,4-dione, (S) -5- (4- Amino-butyl) -1- (2,5-dimethoxy-tetrahydrofuran-3-ylmethyl) -3-ethyl-imidazolidine-2,4-dione, (S) -5- (4-amino -Butyl) -1- (2,2-diphenyl-ethyl) -3-ethyl-imidazolidine-2,4-dione, (S) -5- (4-amino-butyl) -1- (anthracene -9-ylmethyl) -3-ethyl-imidazole Dean-2,4-dione, (S) -5- (4-amino-butyl) -1- (3,5-bis-benzyloxy-benzyl) -3-ethyl-imidazolidine-2,4- Dione, (S) -5- (4-amino-butyl) -3-ethyl-1- (naphthalen-2-ylmethyl) -imidazolidine-2,4-dione, (S) -5- (4 -Amino-butyl) -1- (naphthalen-2-ylmethyl) -3-phenyl-imidazolidine-2,4-dione, (S) -5- (4-amino-butyl) -1- (naphthalene -2-ylmethyl) -3-phenyl-imidazolidine-2,4-dione and (S) -5- (4-amino-butyl) -3- (4-ethoxy-phenyl) -1- ( 4-methoxy-benzyl) -imidazolidine-2,4-dione. The Wang resin-bonded carbamate compound represented by Formula 3 was reacted with diisopropylamine to produce the compound of Formula 4, and then the amino protecting group was removed to prepare the compound of Formula 1, salts and isomers thereof. How to: [Formula 1]

[Formula 3]

[Formula 4]

In the above formula, R ¹ is alkyl, cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl at the aryl moiety, or a heterocycle having a ring containing one or two oxygen atoms, condensed with a carbocyclic ring Arylalkyl, R ² has a straight or branched chain alkyl of 1 to 8 carbon atoms, an aralkyl substituted or unsubstituted at an aryl moiety, or in each case a substituted or unsubstituted aryl or a ring containing one or two oxygen atoms and Condensed heterocyclyl, P represents an amino-protecting group. A composition for preventing coagulation and treating thrombosis, comprising a compound of formula 1 according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient together with a pharmaceutically acceptable carrier.