KR100377555B1 - (s)-3-£4-(amino-hydrazono-methyl)-phenyl|-n-cyclopentyl-n-methyl-2-(naphthalene-2-ylsulfonylamino)-propionamide maleate and solvate thereof - Google Patents

Abstract

PURPOSE: A thrombin inhibitor compound in the form of maleate and a stable solvate thereof are provided to increase the solubility of the compound, thereby improving the bioavailability of the compound. CONSTITUTION: The compound is (S)-3-£4-(amino-hydrazono-methyl)-phenyl|-N- cyclopentyl-N-methyl-2-(naphthalene-2-ylsulfonylamino)-propionamide maleate and a solvate thereof. Particularly, the solvate is dioxane solvate. The compound is prepared by reacting (S)-3-£4-(amino-hydrazono-methyl)-phenyl|-N-cyclopentyl-N-methyl-2- (naphthalene-2-ylsulfonylamino)-propionamide with maleic acid in a solvent, and filtering and drying the product.

Description

(S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide maleate and solvate thereof

The invention provides (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide maleate and The present invention relates to a thrombin inhibitor composition characterized by containing the compound as an active ingredient together with a solvate thereof, a method for preparing the compound, and a pharmaceutically acceptable carrier.

Korean Patent Publication No. 95-43957 discloses (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalene-2- Ilsulfonylamino) -propionamide derivatives are disclosed.

[Formula 1]

This compound is a selective thrombin inhibitor that can be used for oral administration, and particularly can be effectively used for preventing blood clotting and treating various thrombosis.

In general, the conversion of the compound to the salt form may change the physicochemical properties of the compound. In particular, when the solubility is increased by converting to a salt form, it is expected to improve the rate of absorption and thus the bioavailability (see Philip L. Gould, International Jounal of Pharmaceutics, 1986, 33, 201). The compound of Formula 1 is obtained in the form of methylsulfuric acid or hydrochloride in the preparation process, and since these salts can be obtained only by lyophilization or spray drying, they are not easy to prepare, and have high crystallinity and hygroscopicity. There have been difficulties such as keeping the environment dry during storage or formulation process.

Under this technical background, the inventors of the present invention provide the (S) -3- [4- (amino-hydrazono-methyl) -phenyl of Formula 1, which can be effectively used for preventing blood coagulation and treating various thrombosis due to its excellent thrombin inhibitory activity. Intensive study to increase the solubility of -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide to improve its bioavailability, reduce hygroscopicity and have crystallinity As a result, the preparation of the maleic acid salt and its stable solvate form not only achieves this purpose, but also finds it extremely preferable in terms of handling, quality control, and formulation.

Thus, according to the invention (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propion Maleates of amides and solvates thereof, preferably solvates with dioxane.

According to the invention also (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide There is provided a method for preparing maleic acid salts and solvates thereof.

According to the invention there is also a (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalene-2) together with a pharmaceutically acceptable carrier. A thrombin inhibitor composition is provided which contains -ylsulfonylamino) -propionamide maleate or a solvate thereof as an active ingredient.

1 is a free compound of (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide , Dissolution curves of hydrochloride and maleate are shown.

Figure 2 shows (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide methylsulfuric Powder X-ray diffractogram of the acid salt is shown.

Figure 3 Powder of (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide hydrochloride X-ray diffractograms are shown.

Figure 4 shows (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide maleate Powder X-ray diffractograms are shown.

Figure 5 shows (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide maleate di Powder X-ray diffractogram of oxane solvate is shown.

6 shows (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide hydrochloride and male After oral administration of acid salt to a dog, the blood concentration curve of the drug is shown.

7 shows (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino)-formulated into tablets. Elution curve of propionamide maleate is shown.

Maleates and solvates thereof according to the present invention, like other salts, exhibit better solubility than free compounds in pH 3 or higher buffers, and thus exhibit preferred bioavailability over free compounds upon oral administration. However, in addition to such bioavailability, it has been found to be preferable in comparison with other salts in terms of various physical and chemical properties.

First, maleate or its solvate was found to be extremely stable against moisture and hardly absorb moisture in the air. Hygroscopicity of drug substance compounds is a very important factor in terms of the storage of the compound, the cost of the drug formulation process, and the stability and quality control of the drug. That is, in the case of the hygroscopic compound, since the moisture content is easily changed according to the relative humidity, it is necessary to keep the environment dry during the storage or formulation process, and this requires a separate cost. In addition, since the water contained in the additive or the capsule base may be transferred to the raw material during the formulation into tablets or capsules, the type of formulation and formulation additives applicable may be limited and may affect the quality and stability of the drug. . In particular, when the standard or sample for the purpose of quantification is not tested in a dry room, it may cause an error in the experiment and may also cause problems in quality control. As a result of the experiment, hydrochloride and methylsulfuric acid salts are observed to be hygroscopic and change into agglomerates rather than powders when hygroscopic. Therefore, storage and handling in a dry room are absolutely required, and many limitations in terms of formulation are followed. On the other hand, maleic acid according to the present invention was found to have excellent physical properties compared to other salts by maintaining the powder state without prolonged hygroscopic at 93% relative humidity (see Experimental Example 2).

Second, significant differences were found in the crystallinity of the salts. Powder X-ray diffraction experiments confirmed that the maleate of the present invention was crystalline while the other salts were amorphous (see Figures 2-5). Crystallization of salts is of great significance for the production process. The ease of manufacturing process in the bulk synthesis of the drug substance has a significant impact on the overall synthesis cost, because the most preferred form is to filter and dry the salt in the crystalline form at the end of the manufacturing process. The maleic acid salts and solvates thereof of the present invention are easily obtained through filtration and drying processes because they can precipitate out of the crystals from the reaction solution, while the other salts are amorphous and not obtained as crystals in the reaction. Thus, for example, methylsulfuric acid or hydrochloride can only be obtained by lyophilization or spray drying (see Comparative Examples 1, 2, and 4). Lyophilization is long and expensive and spray drying yields. It is not preferable because it has disadvantages such as falling. In short, the maleates and solvates thereof according to the present invention can have great advantages in terms of manufacturing process compared to other salts because they are obtained in crystalline form.

Maleates according to the present invention having such properties can be prepared by reacting a free compound with maleic acid in a suitable solvent and performing a filtration and drying process (see Examples 1 and 2). That is, as mentioned above, the method for preparing maleate does not require a lyophilization process, unlike the method for preparing a corresponding methylsulfuric acid salt or hydrochloride, and can be obtained as a crystalline solid very easily. Formulation is very simple.

Preferred for use as a solvent in the preparation of the maleate according to the present invention is water or a water miscible organic solvent, and a mixed solvent thereof may be used. Maleic acid is preferably used in an amount of 0.9 to 1.1 equivalent times based on the free compound, and the reaction is usually carried out at room temperature.

In order to convert the obtained maleate to its dioxane solvate, for example, the maleate is first added to a mixed solvent of 1,4-dioxane and ethanol (3.1, v / v), dissolved by heating, and concentrated. N-hexane was added to crystallize. Similar methods can be used to prepare solvates other than dioxane solvates.

The present invention also provides (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl- which has such superior advantages in manufacturing process, formulation and storage. Provided is a thrombin inhibitor composition containing 2- (naphthalen-2-ylsulfonylamino) -propionamide maleate or a solvate thereof as an active ingredient and combined with a pharmaceutically acceptable carrier.

The composition according to the present invention may be prepared in consideration of the contents disclosed in the Republic of Korea Patent Publication No. 95-43957 and those commonly known to those skilled in the art. In other words, the composition according to the present invention is formulated into a pharmaceutical preparation in solid, semi-solid or liquid form suitable for oral or parenteral administration by combining a compound of formula 1 with a pharmaceutically acceptable inert carrier upon clinical administration. May be administered. In particular, the composition according to the present invention is preferably formulated in the form of tablets, capsules, etc., which are convenient to administer, and the preparation and pharmacokinetic properties thereof can be referred to the following experimental examples and combination examples.

Pharmaceutically acceptable inert carriers which may be suitably used for this purpose may be solid or liquid and any one or more of a substance which can act as a diluent, flavor, solubilizer, lubricant, suspending agent, binder, tableting agent. Can be. Specific examples of solid or liquid carriers suitable for use in the present invention may include lactose, starch, microcrystalline cellulose, povidone and the like.

In the case of use to exhibit thrombin inhibitory activity, the maleate salt or solvate according to the present invention is preferably at a dosage of 5 to 30 mg per kilogram per day initially based on the active compound. However, the dosage may vary depending on the needs of the patient, the degree of condition to be treated, the compound to be used, and determining the desired dosage in a particular condition is a technique known to those skilled in the art. In general, treatment begins with a dosage less than the optimal amount of the compound. Then increase the dosage in small increments until the optimum effect occurs. For convenience, the total daily dosage may be divided several times and administered throughout the day.

Hereinafter, the present invention will be described in more detail based on the following Comparative Examples, Examples and Experimental Examples. However, these are only for the understanding of the present invention, and the scope of the present invention is not limited by them in any sense.

Comparative Example 1: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide methylsulfur Preparation of furic acid salt

(S) -N-cyclopentyl-3- [4- (imino-methylthio-methyl) -phenyl] -N-methyl-2- (naphthalene-2-sulfonylamino) -propionamide methyl of the formula 364.3 g (0.587 mol) of sulfurates were suspended in 2.3 L of acetonitrile and 30 g (0.588 mol) of 98% H ₂ NNH ₂ H ₂ O was maintained while maintaining the suspension at 0-5 DEG C. Dropped over minutes:

[Formula 2]

The reaction solution was stirred for 2 hours, the insoluble material was filtered off and concentrated under reduced pressure. 1 L of distilled water was added to 0.6 L of the concentrate, and washed twice with 1.5 L of ethyl acetate. The aqueous layer was concentrated to about 1.2 L under reduced pressure and then lyophilized to give 345.7 g (97.4%) of the title compound as a white solid. At this time, the purity was 99% (LC Area%).

Comparative Example 2: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide hydrochloride Manufacture (1)

(S) -N-cyclopentyl-3- [4- (imino-methylthio-methyl) -phenyl] -N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide methylsulfur of formula (2) 131 g (2.56 mole) of 98% -NH ₂ NH ₂ H ₂ O was added dropwise while maintaining a solution of 1.6 kg (2.57 mole) of furic acid salt in 12 L of acetonitrile while being cooled at 5 ° C. The reaction solution was stirred for 1 hour and 46 minutes and nitrogen gas was injected to remove methanethiol (CH ₃ SH) generated during the reaction. Insoluble material was filtered off and concentrated to 3 L under reduced pressure. 5 L of distilled water was added to the concentrate, and the resultant was washed twice with 9 L of ethyl acetate. The organic layer was extracted with 3 L of distilled water.

Each aqueous layer was combined and passed through 6 kg of chloride ion exchange resin (DOWEX 1X8-100) and concentrated to 5.4 L. The concentrate was lyophilized to yield 1.20 kg (yield 88.2%) of the title compound as an off white solid. At this time, the purity was 99% (HPLC Area%).

Comparative Example 3: Preparation of (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide

(S) -N-cyclopentyl-3- [4- (imino-methylthio-methyl) -petyl] -N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide methylsulfur of formula (2) 0.19 kg (3.80 mol) of 98% -NH ₂ NH ₂ .H ₂ O was added dropwise while maintaining a solution of 2.20 kg (3.54 mol) of furic acid salt in 18 L of acetonitrile while being kept cooled at 12 ° C. The reaction solution was stirred for 1 hour and nitrogen gas was injected to remove methanethiol (CH ₃ SH) generated during the reaction. Insoluble matter was filtered off and concentrated to 3 L under reduced pressure. 6 L of distilled water was added to the concentrate, and the resultant was washed twice with 11 L and 4.5 L of ethyl acetate, and the organic layer was extracted with 4 L of distilled water. The distilled water was washed with 10 liters of ethyl acetate, and the aqueous layers were collected and put into a reaction vessel. 11 L of ethanol was added thereto, and an aqueous sodium hydroxide solution (NaOH 0.14 kg + H ₂ O 3.5 L) was slowly added dropwise. After stirring the reaction solution for 2 hours, the resulting solid was filtered and dried with nitrogen gas to give 1.36 kg (yield 77.7%) of the title compound as a white or white solid. At this time, the purity was 99% (HPLC Area%).

Comparative Example 4: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide hydrochloride Manufacture (2)

1.25 kg (2.53 mol) of the compound obtained in Comparative Example 3 was suspended in 7 L of distilled water, and then the reaction solution was cooled to 0 ° C or lower. While adjusting the reaction temperature, 95 g (2.60 mol) of hydrochloric acid gas was blown into the reaction solution. After the reaction solution became clear, lyophilization was carried out to quantitatively obtain 1.34 kg of the title compound as a white solid.

Example 1 (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide maleate Manufacturing of (1)

1.25 kg (2.53 mol) of the compound obtained in Comparative Example 3 and 17 L of ethanol were added to the reaction vessel and stirred at room temperature. 0.29 kg (2.50 mol) of maleic acid was dissolved in 2.8 L of ethanol to form a clear solution, and then added dropwise to the reaction solution for 8 minutes. After stirring for 3 hours, the resulting solid was filtered and washed with 5.6 L of ethanol. Drying with nitrogen gas gave 1.38 kg (yield 89.6%) of the title compound as a white or white solid. At this time, the purity was 99% (HPLC Area%).

Example 2 (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide maleate Manufacturing of (2)

A white-white solid was prepared in the same manner as in Example 1 from 1.25 kg (2.53 mol) of the compound obtained in Comparative Example 3, except that a mixed solvent of ethanol and water (9/1, v / v) was used instead of ethanol. 1.42 kg (yield 92.0%) of the title compound were obtained.

Example 3: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide maleate Preparation of Dioxane Solvate

150 mg of the compound obtained in Example 1 was added to 30 ml of a mixed solvent of 1,4-dioxane and ethanol (3/1, v / v), and then dissolved by heating. The solvent was concentrated to 1/3 and 5 ml of n-hexane was added and left to stand. The resulting solid was filtered and dried to give 128 mg (85.3% yield) of the title compound.

Experimental Example 1: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide free compound Dissolution Test of Hydrochloride, Maleate and Maleate

Elution tests of the free compounds, hydrochloride and maleate were carried out in solutions of pH 1.08 (0.1 normal hydrochloric acid solution), 3.34 (glycine hydrochloric acid buffer solution), and 6.71 (phosphate buffer solution), respectively. Samples were added to 3 ml of each solution in excess, and samples were taken at 15, 35, and 60 minutes while stirring in a 37 ° C thermostat and analyzed by ultraviolet spectroscopy. However, hydrochloride was dissolved more than 500 mg in pH 1.08 solution, so no further experiments were performed.

Each result is shown in FIG. 1, and as can be seen from the results, all the measured samples reached a constant solubility within 15 minutes, and the solubility of hydrochloride and maleate was much higher than the free compound, especially in pH 3.34 and 6.71 solutions. .

Experimental Example 2: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide methylsulfur Hygroscopicity measurement of furic acid salts, hydrochlorides and maleates

Several desiccators were added with saturated aqueous solutions of salts as shown in Table 1 below to adjust the relative humidity inside the desiccators to the values shown in Table 1, followed by hydrochloride, methylsulfuric acid salts, and those obtained in Example 4. The equilibrium water content of maleate was measured at various relative humidity.

TABLE 1

Each hydrochloride and methylsulfuric acid salt was thinned by about 60 mg of petri dish, which was weighed precisely beforehand, and weighed again, and then desiccated at 52, 64, 75% relative humidity among the desiccators shown in Table 1. Each petri dish was put into the tray. The desiccators were left in a 25 ° C. thermostat for 7 days and the samples were taken out and weighed. The weight change is shown in Table 2 below. Regardless of weight change, however, methylsulfuric acid turned into a viscous liquid at over 52% relative humidity and hydrochloric acid into a viscous sample when stored at 75% relative humidity within 2 days of incubation.

On the other hand, the maleate is thinned by about 70 mg of petri dish, stored in a desiccator of 23% relative humidity for 4 days, weighed, and stored in the desiccator of the next step. The weight was again measured and stored in a desiccator having a relative humidity of the next step. In this way, it was observed up to 93% relative humidity. When the weight was extracted from each relative humidity, there was no change in weight. Despite being stored for one month at 93% relative humidity, there was no change in weight and no change in appearance.

TABLE 2

The initial moisture content of hydrochloride, measured by thermogravimetric analysis (TGA) (weight loss up to 100 ° C.), was 1.48% and methylsulfuric acid salt was 2.78%.

Experimental Example 3: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide methylsulfur Powder X-ray Diffraction Test of Puric Acid, Hydrochloride, Maleate and Maleate Dioxane Solvate

40 mg each of hydrochloride, methylsulfuric acid salt, maleic acid salt obtained in Example 4, and maleic acid dioxane solvate obtained in Example 6 were applied thinly to a sample holder, and then powder X-ray under the conditions as shown below. Diffraction analysis (35 kV × 20 mA Rigaku GeigeflexD / max-III C) was performed.

The powder X-ray diffraction analysis results of hydrochloride, methylsulfuric acid salt, maleate and maleic acid dioxane solvate are shown in Figs. 2 to 5, respectively. It can be seen that the maleate dioxane solvate has crystallinity.

Experimental Example 4: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide hydrochloride and Oral Absorption Test of Male Maleates

Fill the gelatin capsules with powders of hydrochloride and maleate, orally administer the dog at a 30 mg / kg dose and collect blood at 0, 10, 20, 40, 60, 90, 120, 180, 240, 360, 480 minutes. It was. After the collected blood was treated with protein, the concentration of drug in the blood was measured by high performance liquid chromatography. The change in blood drug concentration over time is shown in FIG. 6, and the pharmacokinetic parameters calculated from the results are shown in Table 3 below. As a result, the maximum blood concentration time of maleate was slightly slower than hydrochloride, but there was no significant difference between the two salts in the degree of absorption.

TABLE 3

Formulation Example 1: (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide maleate Manufacture of tablets

A powder mixture was prepared with the composition as shown in Table 4 below to make 250 mg tablets (diameter 9 mm) per tablet using a KORSCH automatic tablet press.

TABLE 4

The hardness of the tablets thus prepared was measured (using the Euweka TBH 28 instrument), and the results measured in Newtonian units for each of the six tablets were 47, 52, 46, 51, 44, and 52, respectively. In addition, the result of the dissolution test in the 100mM phosphate buffer solution (pH 6.82) for this tablet is shown in Figure 7, it can be seen that 82% of the active ingredient is eluted within 6 minutes.

Claims (6)

Maleate and solvent thereof of (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide freight. The compound of claim 1 which is a dioxane solvate. (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino)-with a pharmaceutically acceptable carrier A thrombin inhibitor composition comprising propionamide maleate or a solvate thereof as an active ingredient. Reaction of (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino) -propionamide with maleic acid in a solvent And a filtration and drying process to prepare the maleate as defined in claim 1 and solvates thereof. The method according to claim 4, wherein the solvent is water or a water miscible organic solvent, or a mixed solvent thereof. The maleic acid of claim 4, wherein the maleic acid is (S) -3- [4- (amino-hydrazono-methyl) -phenyl] -N-cyclopentyl-N-methyl-2- (naphthalen-2-ylsulfonylamino). -Using in an amount of 0.9 to 1.1 equivalent times based on propionamide.