KR100350737B1 - 2-Chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione Derivatives - Google Patents

Abstract

The present invention provides a novel 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative of the general formula (I) having a platelet aggregation inhibitory activity, a method for preparing the same and an active ingredient thereof It relates to platelet aggregation inhibitors.

Where

R ₁ , R ₂ , R ₃ are the same or different from each other hydrogen (H), fluorine (F), chloro (Cl), bromine (Br), acet (CH ₃ CO), nitro (NO ₂ ), trifluorine Romoxy (CF ₃ ) group or ethylcarboxy (C ₂ H ₅ OCO) group.

The present invention provides 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivatives and a method for preparing the same, and 2-chloro-3-phenylamino-5-hydroxy- according to the present invention. The 1,4-naphthalenedione derivative has a platelet aggregation inhibitory effect and can be used as a clinically useful platelet aggregation inhibitor.

Description

2-Chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative {2-Chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione Derivatives}

The present invention relates to novel 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivatives. More specifically, the present invention provides a novel 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative of the general formula (I) having a platelet aggregation inhibitory activity, a method for preparing the same It relates to a platelet aggregation inhibitor and a thrombosis therapy comprising as an active ingredient.

Where

R ₁ , R ₂ , R ₃ are the same or different from each other hydrogen (H), fluorine (F), chloro (Cl), bromine (Br), acet (CH ₃ CO), nitro (NO ₂ ), trifluorine Romoxy (CF ₃ ) group or ethylcarboxy (C ₂ H ₅ OCO) group.

Platelets make up a small portion of the blood constituent cells, but they respond easily to minute stimuli and play an important role in maintaining homeostasis in the body, such as blood coagulation. In addition, abnormalities in platelet aggregation ability caused by a change in platelet function may cause a hemostatic mechanism or a microcirculation disorder. In particular, blood disorders that frequently occur in adult diseases such as hypertension and arteriosclerosis, and metabolic diseases such as diabetes, which are rapidly increasing in modern society, are also reported to be due to an ideal hyperactivity of platelet aggregation ability. It is anticipated that normalization of these diseases can treat or prevent these related diseases.

On the other hand, the generation of a thrombus (thrombus) is a natural defense means to protect the living body by hemostasis that platelets are activated and adhesion and aggregation occurs when bleeding due to blood vessel damage. However, when platelets are activated excessively due to pathological factors such as aging, intravascular aggregation causes thrombosis, which impedes blood circulation and causes major causes of adult diseases such as myocardial infarction, arteriosclerosis, hypertension, and diabetes. Becomes Therefore, much research has been made on the development of platelet aggregation inhibitors for preventing thrombosis.

Generally, 1,4-naphthalenedione derivatives have various pharmacological actions such as anticoagulant action, antiplatelet action, antibacterial action and antifungal action, and are used as important substances for the development of new drugs such as antibacterial agents.

According to the results of various researchers, 1,4-naphthalenedione derivative vitamin K ₃ (menadione) is reported to be effective in inhibiting the aggregation of human platelets, the mechanism of action is not yet known. (Blackwell, GJ, et al., Thromb. Res., 37, 103-114, 1985).

On the other hand, 2-chloro-3-methyl-1,4-naphthalenedione (CMN) has been shown to inhibit the platelet aggregation reaction in rabbits, which is responsible for the metabolism of phospholipids by various platelet activists. It has been reported to appear because -chloro-3-methyl-1,4-naphthalenedione inhibits (see Ko, FN, et al., Thromb. Res., 57, 453-463, 1990). In addition, 5,8-dihydroxy-1,4-naphthalenedione derivatives (cyconin derivatives) have been shown to inhibit platelet aggregation reactions (CA Vol. 105, 743p, 226090C; JP (Sho) 61-). 143,334), 2-alkyl / arylcarboxamido-3-chloro-1,4-naphthalenedi temperature and exhibited good platelet aggregation inhibitory activity (Lien, JC, et al., Chem. Pharm. Bull., 44 ( 6), 1181-1187, 1996; Lien, JC, Huang, et al., Bioorg.Med. Chem., 5 (12), 2111-2120, 1997), 2-chloro-3- (N-4-sia Nophenyl-amido) -1,4-naphthalenedione is known to exert platelet aggregation inhibitory effects by inhibiting the production of thromboxane A2 and granule release, which play the most important role in platelet aggregation (Chang, Chang). , TS, et al., Biochem.Pharmacol., 54, 259-268, 1997).

Accordingly, the present inventors have made diligent efforts to develop new compounds exhibiting platelet aggregation inhibitory effects, and as a result, a novel 1,4-naphthalenedione derivative, 2-chloro- 3-phenylamino-5-hydroxy-1,4-naphthalenedione Synthesis of a variety of derivatives, and confirmed that the electrical substances exhibit excellent platelet aggregation inhibitory effect, to complete the present invention.

After all, the main object of the present invention is to provide 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivatives.

Another object of the present invention is to provide a method for preparing the above 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative.

Still another object of the present invention is to provide a thrombosis therapeutic composition containing 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative as an active ingredient.

The present invention relates to 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative (I) exhibiting platelet aggregation inhibitory effect.

Where

R ₁ , R ₂ , R ₃ are the same or different from each other hydrogen (H), fluorine (F), chloro (Cl), bromine (Br), acet (CH ₃ CO), nitro (NO ₂ ), trifluorine Romoxy (CF ₃ ) group or ethylcarboxy (C ₂ H ₅ OCO) group.

As a specific example of the compound represented by general formula (I) which concerns on this invention,

(1) 2- (4-acetophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (hereinafter referred to as 'PRCK121');

(2) 2- (4-chloro-3-nitrophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (hereinafter referred to as 'PRCK140');

(3) 2- (4-trifluoromethoxyphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (hereinafter referred to as 'PRCK144');

(4) 2- (2-trifluoromethoxyphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (hereinafter referred to as 'PRCK145');

(5) 2- (4-methyl-3-nitrophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (hereinafter referred to as 'PRCK163');

(6) 2- (4-ethylcarboxyphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (hereinafter referred to as 'PRCK169');

(7) 2- (2-chloro-4-bromophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (hereinafter referred to as 'PRCK174'); And,

(8) 2- (2-bromo-4-fluorophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (hereinafter referred to as 'PRCK178')

Etc. can be mentioned.

The novel 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative (I) of the present invention is a 2,3-dichloro-5-hydroxy-1,4- of compound (II). Naphthalenedione can be prepared by reacting with amines of the general formula (III), respectively.

Where

R ₁ , R ₂ and R ₃ are as defined above.

2,3-dichloro-5-hydroxy-1,4-naphthalenedione of Compound (II) used as starting material in the preparation method of the present invention is a compound known in the art, and 5-hydroxy-1, It can be conveniently prepared by reacting 4-naphthalenedione with chlorine (Cl ₂ ) gas. As the amines of the general formula (III), commercially available ones are used.

According to the preparation method of the present invention, the compound of formula (I) is prepared by reacting compound (II) with an amine derivative such as alkyl or arylamine of formula (III) in an alcohol or dimethyl sulfoxide (DMSO) solvent. . At this time, the alcohol used as a solvent is preferably a lower alkanol having 1 to 3 carbon atoms such as methanol, ethanol or isopropanol, but in particular, ethanol is preferable, and the reaction temperature is generally room temperature to 100 ° C, preferably the solvent used. It can be carried out under reflux at the boiling point of. The reaction time in the present invention is generally 2 to 6 hours, preferably 3 to 5 hours. After completion of the reaction, the compound of formula (I) obtained can be separated and purified by conventional post-treatment methods such as recrystallization or chromatography, if necessary.

The compound of general formula (I) according to the present invention has a platelet aggregation inhibitory effect and can be used as a clinically useful platelet aggregation inhibitor and therapeutic agent for thrombosis.

Platelet aggregation ability is known to be greatly affected by processes such as secretion ability from platelet granules, release of arachidonic acid in platelets and metabolism. In other words, platelets contain more granules than other cells, such as dense granules, alpha-granules, lysosomes and peroxisomes. Very useful substances are stored. When platelets are activated by a stimulus, the substances stored in the granules are secreted out of the cells. In particular, dense granules store adenine nucleotides such as ADP and ATP, calcium, and serotonin, which are released when platelets aggregate and cause irreversible secondary aggregation. That is, the irreversible secondary aggregation reaction in the case of inducing aggregation with ADP or thrombin can be regarded as the result of the granule secretion. In addition, thromboxane A ₂ produced by arachidonic acid metabolism in platelets is known to be a potent platelet aggregation agent, and is known to play a pivotal role in platelet aggregation.

Hereinafter, the preparation method of the 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative represented by the general formula (I) and the platelet aggregation inhibitory effect will be described in more detail. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example 1 Preparation of Compound (II)

20 g of 5-hydroxy-1,4-naphthalenedione was dissolved in 200 ml of acetic acid, and about 10 g of chlorine (Cl ₂ ) gas was injected into the resulting solution, followed by shaking for 4 hours at room temperature. The reaction mixture was then poured into 500 g of ice and the resulting precipitate was filtered off and dried in air. The precipitate was suspended in ethanol and then heated in a water bath for 30 minutes and left at room temperature to give 25.0 g of the desired 2,3-dichloro-5-hydroxy-1,4-naphthalenedione (II) (yield 69 %).

Example 2 Preparation of Compound (I)

In a 200 ml round bottom flask, 1 g of 2,3-dichloro-5-hydroxy-1,4-naphthalenedione obtained in Example 1 was dissolved in 50 ml of an aqueous 95% ethanol solution, and 0.6 ml of amine was added thereto, respectively. The reaction mixture was reacted by refluxing under heating for 3 hours. After confirming the completion of the reaction by thin layer chromatography, the reaction solution was cooled and the precipitated crystals were filtered and recrystallized from ethanol to purify each of the following desired title compounds PRCK121, PRCK140, PRCK144, PRCK145, PRCK163, PRCK169, PRCK174 and PRCK178 was obtained. The results for the above title compound are as follows.

(1) 2- (4-acetophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (PRCK 121):

mp 208-210 ° C. (dark brown powder); IR (KBr, cm ⁻¹ ): 3300 (NH), 1640 (C═O), 1350 (SCOCH ₃ ), 1590, 1450, 1350, 1220; ¹ H-NMR (DMSO-d ₆ ): δ 2.65 (s, 3H, CH ₃ ), 7.30-7.48 (4H, m, benzene ring), 7.70-7.85 (3H, m, 6,7,8 H), 9.80 (1H, s, NH), 12.36 (1H, s, OH); MS (m / z): 341 (M ⁺ ), 326 (M-15), 299, 292, 263, 262, 207, 206, 146, 120

(2) 2- (4-chloro-3-nitrophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (PRCK140):

mp 145-146 ° C. (bright brown powder); IR (KBr, cm ⁻¹ ): 3300-2500 (OH), 3240 (NH), 1660 (C═O), 1430, 1270, 1220, 750; ¹ H-NMR (DMSO-d ₆ ): δ 7.40-7.50 (3H, m, benzene ring), 7.80-7.90 (3H, m, 6,7,8 H), 10.02 (1H, s, NH), 12.90 (1H, s, OH); MS (m / z): 378 (M ⁺ ), 377 (M-1), 343 (M-35), 331, 297, 296, 241, 207, 99

(3) 2- (4-trifluoromethoxyphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (PRCK144):

mp 175-180 ° C. (dark brown powder); IR (KBr, cm ⁻¹ ): 3220 (s, NH), 3300-2650 (s, OH), 1670 (s, C═O), 1500, 900, 800; ¹ H-NMR (DMSO-d ₆ ): δ 7.20-7.30 (4H, m, benzene ring), 7.40-7.50 (3H, m, 6,7,8 H), 9.70 (1H, s, NH), 12.40 (1H, s, OH); MS (m / z): 383 (M ⁺ ), 382 (M-1), 348 (M-35), 332, 298, 228, 207, 121

(4) 2- (2-trifluoromethoxyphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (PRCK145):

mp 94 ° C. (yellow brown powder); IR (KBr, cm ⁻¹ ): 3210 (s, NH), 1670 (s, C═O), 1630, 1440, 1320, 1200, 890, 730; ¹ H-NMR (DMSO-d ₆ ): δ 7.50-7.73 (4H, m, benzene ring), 7.80-7.90 (3H, m, 6,7,8 H), 9.70 (1H, s, NH), 12.00 (1H, s, OH); MS (m / z): 367 (M ⁺ ), 332 (M-35), 312, 280, 276, 241, 207, 179, 157, 87

(5) 2- (4-methyl-3-nitrophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (PRCK163):

mp 192-194 ° C (red powder); IR (KBr, cm ⁻¹ ): 3300 (NH), 1680 (C═O), 1620, 1550, 1480, 1370, 1230; ¹ H-NMR (DMSO-d ₆ ): δ 3.42 (3H, s, CH ₃ ), 7.40-7.50 (3H, m, benzene ring), 7.80 (3H, m, 6,7,8 H), 9.90 ( 1H, s, NH), 13.00 (1H, s, OH); MS (m / z): 358 (M ⁺ ), 342 (M-15), 323 (M-35), 313, 311, 276, 89

(6) 2- (4-ethylcarboxylphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (PRCK169):

mp 242-245 ° C. (brown plate); IR (KBr, cm ⁻¹ ): 3260 (NH), 2980 (−CH), 1720 (C═O), 1170, 1670 (S, COO), 1620, 1600, 1500, 1180, 700; ¹ H-NMR (DMSO-d ₆ ): δ 1.41 (3H, t, J = 7.5 Hz, CH ₃ ), 4.40 (2H, Q, J = 15 Hz COO-CH _2- ), 7.20-7.60 (4H, m, benzene ring), 7.90-8.00 (3H, m, 6,7,8 H), 9.70 (1H, s, NH), 12.40 (1H, s, OH); MS (m / z): 371 (M ⁺ ), 360, 326, 298, 290, 264, 263, 235, 207, 145

(7) 2- (2-chloro-4-bromophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (PRCK174):

mp 93.5 ° C. (shining orange green powder); IR (KBr, cm ⁻¹ ): 3856, 3366 (NH), 2366, 1203 (CN), 889; ¹ H-NMR (DMSO-d ₆ ): 6.82-7.50 (3H, m, benzenering), 7.53-7.92 (3H, m, C ₆ H ₃ OH), 11.48 (1H, s, OH); MS (m / z): 378 (M ⁺ ), 376 (M-1), 242, 207

(8) 2- (2-bromo-4-fluorophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione (PRCK178):

mp 110.0 ° C. (dark orange powder); IR (KBr, cm ⁻¹ ): 3857, 3315 (NH), 1494, 1204 (CN), 733; ¹ H-NMR (DMSO-d ₆ ): 6.88-7.41 (3H, m, benzene ring), 7.49-7.93 (3H, m, C ₆ H ₃ OH), 11.48 (1H, s, OH), 9.27 (1H , s, NH); MS (m / z): 397 (M ⁺ ), 316 (M-80), 288

Example 3 Measurement of Inhibitory Effect on Platelet Aggregation Ability of Platelet Activator

(1) Preparation of plasma and washed platelets containing a large amount of platelets:

Using a syringe preloaded with sodium citrate (final concentration 0.38%), blood from a healthy adult man with normal platelet function was centrifuged at 160Xg for 15 minutes at room temperature, and plasma containing a large amount of platelets from the supernatant was obtained. Obtained. The precipitate obtained by centrifuging the platelet-rich plasma for 15 minutes again at 1500Xg was washed with a wash buffer containing 2 mM ethylenediamine tetraacetic acid (129 mM sodium chloride, 0.8 mM potassium diphosphate, 8.9 mM sodium carbonate, 2.8 mM potassium chloride, 0.8 mM). Magnesium chloride, 5.6 mM glucose, 10 mM N- [2-hydroxyethyl] -piperazine-N '-[2-ethanesulfonic acid], the pH of the buffer solution 7.3), washed with protein and calcium in plasma After removal, modified tierod buffer (129 mM sodium chloride, 0.8 mM potassium phosphate, 8.9 mM sodium carbonate, 2.8 mM potassium chloride 0.8 mM magnesium chloride, 5.6 mM glucose, 10 mM N-) containing 0.35% bovine serum albumin Washed platelets were prepared by carefully resuspending in [2-hydroxyethyl] piperazine-N '-[2-ethanesulfonic acid], buffer acidity pH 7.4).

(2) Determination of platelet aggregation ability:

The platelet aggregation ability was measured using a platelet aggregation ability measuring instrument (Chronolog, USA) according to the method of Born et al. (Born, GVR, Nature, 194, 927, 1962). Judgment was made by measuring the change of. In contrast to the degree of light passing in the case of little platelets, 495 μl of plasma containing a large amount of platelets prepared in (1) was pretreated at 37 ° C. for 30 seconds, and then dissolved in dimethyl sulfoxide. Each compound (optimum concentration 10 ^-4 M) according to the invention was added and reacted again at 37 DEG C for 8 minutes, and then 2.5 mu l of thrombin (final concentration 0.5 U / ml) was added to change the degree of light passing through for 10 minutes. Was measured during. In addition, 495 μl of the washed platelets prepared in the above (1) were pretreated at 37 ° C. for 30 seconds, and then dissolved in dimethyl sulfoxide. 2.5 μl of the compound was added and reacted again at 37 ° C. for 8 minutes. Then, 2.5 μl of platelet activators (10 μg / ml of collagen, or 2 μM of ADP) was added to cause aggregation of platelets, and the change in the degree of light passage for 10 minutes was measured. Experimental results were determined by calculating the degree of light passing through the platelet activating material as 100%, and calculating the degree of light passing in each case as a relative analogy.

At this time, using 0.02M dimethyl sulfoxide instead of the test substance as a control, the cohesion of the control was set to 100% and the inhibition rate for each concentration of the drug was calculated, and then the IC ₅₀ value of 50% inhibition rate of each test substance was determined. . The extent to which platelet aggregation was inhibited by the drug was calculated as follows.

A-B

Inhibition% = ----- X 100

A

A: control aggregation (%)

B: Coagulation at the time of treatment (%)

IC ₅₀ (concentration of drug that inhibits platelet aggregation 50%) was calculated by the Probit method.

Inhibitory Effect of Compounds of the Invention on Platelet Aggregation Ability Activated by Collagen and ADP sample ADP 20μM Collagen 200 μg / ml Cohesion % Inhibition Cohesion % Inhibition PRCK121 22 78 95 5 PRCK140 16 84 14 86 PRCK144 6 94 11 89 PRCK145 One 99 11 89 PRCK163 45 55 75 25 PRCK169 66 66 7 93 PRCK174 9 84 8 90 PRCK178 12 80 7 92

50% inhibition (IC ₅₀ ) of ADP-activated platelet aggregation ability of the compound of the present invention compound IC ₅₀ (μM) PRCK121PRCK140PRCK144PRCK145PRCK163PRCK169PRCK174PRCK178 aspirin 41.527.229.441.837.416.225.226.3148.7

As can be seen from the results of Table 1 and Table 2, the effects of the compounds of the present invention PRCK121, PRCK140, PRCK144, PRCK145, PRCK163, PRCK169, PRCK174 and PRCK178 on the aggregation ability of the platelet activator collagen, ADP to wash platelets In each case, it showed better anti-aggregation effect than the control group (Table 1). Each concentration was found to be 41.5, 27.2, 29.4, 41.8, 37.4, 16.2, 25.2, and 26.3 μM (Table 2), and the compounds of the present invention all confirmed to inhibit platelet aggregation by various platelet activators. It became.

Example 4 : antithrombosis test

Induction of experimental thrombi was carried out according to the method of Dimino et al. (See Dimino, G. and MJ Silver., J. Pharmacol.Exp. Ther., 225, 57-60, 1983). At this time, the animals used were male ICR mice weighing about 20 to 25 g fasted overnight. The induction of thrombus was caused by the injection of platelet aggregation reagents (collagen, epinephrine) into the tail vein so that blood clots were generated in the lung microvessels. In order to determine the antithrombotic effect of the PRCK family, the injection of the tail vein at a dose of 200 μl per mouse was performed. PRCK121 (27 mg / kg, 135, 1 hour before intravenous injection of platelet aggregation reagent) mg / kg) and PRCK140 (26.7 mg / kg, 133.7 mg / kg) were suspended in corn oil and orally administered 1 ml each. As a control, 20 mg / kg of aspirin was suspended orally in 0.5% carboxymethylcellulose (CMC). The antithrombotic effect of the PRCK family was calculated as a percentage of the number of experimental animals protected from paralysis or death of mouse hind limbs caused by the administration of platelet aggregation reagents. It was based on time duration and significance test was used with X ² -test.

Effects on Pulmonary thrombosis in rats the compounds of the invention ^‡ sample Dose (mg / kg) Number of dead or paralyzed / experimental animals % Protection Control 0.5% CMC 18/21 - aspirin 50 6/19 ^*** 63.1 PRCK121 300 8/20 ^*** 75.5 60 4/19 ^** 53.3 PRCK140 300 7/20 ^*** 59.2 60 7/18 ^** 54.6

^• PRCK121 and PRCK140 were orally administered 90 minutes prior to intravenous injection of epinephrine (13.2 μg / mouse) and collagen (114 μg / mouse).

Significant difference from control: ^** P <0.01, ^*** P <0.001.

As can be seen from the results of Table 3, 18 of 21 rats were killed or paralyzed in the control group, resulting in thrombosis of 85.7%, and there was a dose-dependently significant antithrombotic effect in the PRCK121 and 140 series administration groups. In the case of PRCK121, 53.3% at 60mg / kg, 75.5% at 300mg / kg, 54.6% at 60mg / kg and 59.2% at 300mg / kg for PRCK140. On the other hand, aspirin, a positive control, showed an inhibitory effect of 63.1% at 50 mg / kg.

Example 5 Antiplatelet Aggregation Activity Test in Ex vivo Conditions

The experimental animals used male Sprague-Dawley rats weighing about 320 to 440 g overnight fasted. Each group was divided into 6-9 rats, and the test substance and aspirin were suspended in 0.5% CMC and administered orally by 1 ml. After 1 hour of oral administration, anesthesia was rapidly anesthetized with ether, using an 18 gauge needle, and the blood sample was collected so that the ratio of 2.2% sodium citrate: blood was 1: 6 (v / v). rich plasma; platelet rich plasma). The supernatant was taken and tested for antiplatelet aggregation activity with collagen and ADP.

Inhibitory effect of oral administration of a compound of the present invention on platelet aggregation in rats ^‡ sample ADP (1.3 μM) Collagen (32.79 μg / ml) Coagulation (%) control(%) Coagulation (%) control(%) Control 36 ± 2 - 53 ± 5 - PRCK121 23 ± 2 ^* 36 ^* 39 ± 1 ^* 26 ^* PRCK140 25 ± 1 ^* 31 ^* 40 ± 3 ^* 25 ^*

^• The compound of the present invention was administered orally, and platelet aggregation was promoted by ADP (1.3 μM) or collagen (32.79 μg / ml).

*: Significant difference from control group (p <0.01)

Mean ± standard deviation, number of animals used per group: 5-7

The results of the experiment are shown in Table 4. As can be seen from the results of Table 4, the result of platelet aggregation inhibition test in ex vivo condition did not show platelet aggregation inhibitory effect in ADP (1.3 μM) and collagen (32.79 μg / ml) in the control group, but PRCK121 and 140 In the case of ADP, 36 and 31% of platelet aggregation inhibitory effect occurred, and collagen had 26 and 25% platelet aggregation inhibitory effect.

Therefore, it was confirmed that the compound of the present invention has a platelet aggregation inhibitory effect through the above various tests.

Example 6 : Acute Toxicity Test

The compound of the present invention was intravenously administered to 10 SD rats for 7 days in an amount of 2850 mg / kg, about 1,000 times the effective amount, but no case was recognized (Table 5). As a result of the weight measurement, both the male and female groups showed no difference compared to the control group (Table 6). As a result, in the acute oral toxicity test of mice of the present invention, no toxicities were observed in the general condition, weight change and autopsy findings.

As described above, the target compound of formula (I) according to the present invention exhibits platelet aggregation inhibitory action, and thus can be used as a clinically useful platelet aggregation inhibitor and thrombosis therapeutic agent. In clinical use of the compound (I) according to the present invention, the active compound may be prepared according to conventional methods using a pharmaceutically acceptable carrier, such as conventional agents in the pharmaceutical field, such as tablets, capsules, solutions, Injectables, ointments, creams, etc. in the form of oral preparations such as suspensions, injectable solutions or suspensions, or dry powders for injection, which are ready-to-use, which can be reconstituted with injectable distilled water at the time of injection, It can be formulated into various formulations such as topical formulations such as liquids.

Carriers used for this purpose are customary in the pharmaceutical art, for example in the case of oral preparations, binders, suspending agents, disintegrants, excipients, solubilizing agents, dispersants, stabilizers, suspending agents, pigments, flavorings In the case of injections, there are preservatives, analgesic agents, solubilizers, stabilizers and the like, and in the case of topical administration agents, there are bases, excipients, lubricants, preservatives and the like. The pharmaceutical preparations thus prepared may be administered orally or parenterally, for example, intravenously, subcutaneously, intraperitoneally, or topically. In addition, in order to prevent the decomposition of the drug by gastric acid during oral administration, it may be desirable to use an antacid in combination, or to formulate an oral administration such as tablets into an enteric coating.

The dosage of the pharmaceutical preparations according to the present invention varies depending on the type of disease to be treated or prevented, the method of administration, the age of the patient, the treatment time, and the like. The dosage should be set within the range of 2.5 to 100 mg / kg based on the active ingredient, and in the case of oral administration within the range of 25 to 500 mg / kg.

As described and demonstrated in detail above, the present invention provides a 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative and a preparation method thereof. The 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative according to the present invention has a platelet aggregation inhibitory effect and can be used as a clinically useful platelet aggregation inhibitor and a thrombosis therapeutic agent.

Claims (7)

delete 2-Chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative compound, characterized in that it is selected from the group consisting of: 2- (4-acetophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione; 2- (4-chloro-3-nitrophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione; 2- (4-trifluoromethoxyphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione; 2- (2-trifluoromethoxyphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione; 2- (4-methyl-3-nitrophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione; 2- (4-ethylcarboxyphenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione; 2- (2-chloro-4-bromophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione; And, 2- (2-Bromo-4-fluorophenyl) -amino-3-chloro-5-hydroxy-1,4-naphthalenedione. delete delete delete delete A platelet aggregation inhibitor comprising the 2-chloro-3-phenylamino-5-hydroxy-1,4-naphthalenedione derivative of claim 2 as an active ingredient and a pharmaceutically acceptable carrier.