Crystalline forms of pyrimidine-4-carboxamide compounds and oral pharmaceutical preparations containing the same

Abstract

The present invention relates to a stable novel crystalline form of the compound of Chemical Formula 1 and a method for obtaining such a crystalline form. The present invention also relates to a pharmaceutical formulation, particularly an oral pharmaceutical formulation, which contains the compound of Chemical Formula 1 (preferably the crystalline form of the compound according to the present invention) as an active ingredient and has ensured stability, high dissolution rate, high bioavailability, etc., and a method for preparing the same.

Description

[Technical Field]

[0001] The present invention relates to new crystalline forms of pyrimidine-4-carboxamide compounds having specific structures and / or oral (pharmaceutical) formulations containing such crystalline solid form drug substances as active ingredients.

[0002] This application claims priority based on Korean Patent Application No. 10-2022-0102292, filed on August 16, 2022, the entire contents of which are incorporated herein by reference in their entirety in the specification and drawings thereof. [Background technology]

[0003] 2-((R)-4-(2-fluoro-4-(methylsulfonyl)phenyl)-2-methylpiperazin-1-yl)-N-((1R,2s,3S,5S,7S)-5-hydroxyadamantan-2-yl)pyrimidine-4-carboxamide), represented by the following chemical formula 1, is a compound included in the chemical formula disclosed in U.S. Patent No. 9,096,571.

[0004] [Chemical formula 1] TIFF0007822103000001.tif35170 The compound of Chemical Formula 1 is effective in preventing or treating non-alcoholic steatohepatitis, and has the advantages of improving dyslipidemia, protecting hepatocytes, and exhibiting anti-fibrotic function (see Korean Patent Registration No. 10-2177304).

[0005] The compound of Chemical Formula 1 has a water solubility of 1 μg / mL or less and is a substance that belongs to the “practically insoluble” category, and is classified as Class 2 according to the Biopharmaceutics Classification System (BCS).

[0006] There are various difficulties in developing drugs using poorly soluble pharmaceutical ingredients. For example, amorphous or partially crystalline ingredients can be used to improve aqueous solubility, but amorphous or partially crystalline ingredients have the disadvantage of being less stable than crystalline ingredients. Furthermore, amorphous or partially crystalline ingredients may undergo crystal transformation during storage or during the drug manufacturing process, making it difficult to control drug dissolution and blood concentration.

[0007] Meanwhile, the preparation of amorphous solid dispersions using hydrophilic polymers and / or water-soluble carriers can be effective in improving drug solubility. However, the amorphous active ingredient in solid dispersions can transition to a crystalline form depending on storage conditions, resulting in changes in dissolution. Furthermore, amorphous solid dispersion formulations have a fundamental problem: their thermodynamic stability is inferior to that of formulations using crystalline raw materials. Summary of the Invention [Problem to be solved by the invention]

[0008] An object of the present invention is to provide a novel crystalline form of the compound of Formula 1 having physicochemical properties useful as an active ingredient in pharmaceuticals, and a method for preparing the crystalline form.

[0009] Another object of the present invention is to provide a pharmaceutical formulation, preferably an oral pharmaceutical formulation, which contains the compound of Formula 1 as an active ingredient, and has ensured stability and a high dissolution rate, and a method for preparing the same. [Means for solving the problem]

[0010] In order to achieve the above object, one embodiment of the present invention is a powder X-ray diffraction (PXRD) analysis in which 2θ diffraction angles are 5.3±0.2°, 7.5±0.2°, 10.1±0.2°, 10.6±0.2°, 11.5±0.2°, 14.3±0.2°, 14.7±0.2°, 15.0±0.2°, 15.5±0.2°, 16.0±0.2°, 16.3±0.2°, 16.5±0.2°, 19.1±0.2°, 19.3±0.2°, 19.7±0.2°, 20.3±0.2°, 20.6±0.2°, 21. Provided is a crystalline form of the compound of Formula 1 (2-((R)-4-(2-fluoro-4-(methylsulfonyl)phenyl)-2-methylpiperazin-1-yl)-N-((1R,2s,3S,5S,7S)-5-hydroxyadamantan-2-yl)pyrimidine-4-carboxamide), having a powder X-ray diffraction pattern with peaks at 4±0.2°, 21.8±0.2°, 22.6±0.2°, and 27.5±0.2°.

[0011] [Chemical formula 1] TIFF0007822103000002.tif35170

[0012] In another aspect of the present invention, there is provided a crystalline form of the compound of Formula 1 having the X-ray diffraction pattern shown in FIG.

[0013] In another aspect of the present invention, the present invention provides a method for producing a compound having a molecular weight of 1100 cm by infrared absorbance analysis. -1 (aromatic CF), 1518cm -1 (NH), 1579cm -1 (aromatic C=C), 1680cm -1 (C=O), 2911 cm -1 (aliphatic CH) and 3445 cm -1 In yet another aspect of the present invention, the present invention provides a crystalline form of the compound of Formula 1, which has an absorbance pattern as shown in Figure 2 in infrared absorbance analysis.

[0014] Even compounds with the same chemical formula exhibit different physicochemical properties depending on the crystalline form. Differences between crystalline forms include, but are not limited to, stability (e.g., heat or light stability), compressibility and density (important for formulation and manufacturing), and dissolution rate (which can affect bioavailability). In particular, differences in stability can cause changes in chemical reactivity (e.g., discoloration, moisture absorption) or mechanical properties (e.g., changes due to conversion to a thermodynamically more stable polymorph). Specifically, crystalline form can affect humidity stability, the ability to form a dissolved compound, and ease of filtration or cleaning.

[0015] As a result of extensive research, the present inventors have secured a stable crystalline form of the compound of Formula 1 according to the present invention, which exhibits desirable physicochemical and mechanical properties in the various aspects described above. In particular, the crystalline form according to the present invention is highly stable, and there is no risk of the physicochemical properties of the active ingredient changing during storage of raw materials, production of pharmaceuticals, storage and distribution of pharmaceuticals.

[0016] In addition, another aspect of the present invention provides a method for preparing a crystalline form of the compound of Formula 1 according to the present invention, comprising: (S1) dissolving the compound of Formula 1 in ethyl acetate; (S2) adding t-butyl methyl ether to the ethyl acetate solution of Step S1 to cause crystallization; and (S3) filtering the suspension of Step S2 to obtain the crystalline compound of Formula 1.

[0017] The present inventors also attempted to prepare an oral formulation using the compound of Chemical Formula 1, particularly the crystalline solid of the present invention, as a pharmaceutical raw material. The compound of Chemical Formula 1 has a water solubility of 1 μg / mL or less and is a substance that belongs to the category of "practically insoluble," and is a poorly soluble substance classified as Class 2 (high permeability, low solubility) according to the Biopharmaceutics classification system (BCS).

[0018] The present inventors have made various attempts to increase the bioavailability of such poorly soluble substances. Previously, the compound of Chemical Formula 1 did not have the property of improving solubility depending on pH, so there was no room for adjusting the drug dissolution rate by creating a salt. Furthermore, using amorphous raw materials or solid dispersion technology in which the active ingredient exists in an amorphous state made it difficult to achieve long-term stability. Furthermore, simply mixing hydrophilic ingredients (e.g., hydrophilic polymers, surfactants) made it difficult to achieve the desired level of bioavailability.

[0019] In this context, the present inventors have confirmed that when a pharmaceutical formulation is prepared from particles prepared by drying water in which the compound of Formula 1 is suspended and a hydrophilic polymer is dissolved, not only can (long-term) stability be achieved but also bioavailability can be dramatically improved, thereby completing the present invention.

[0020] Therefore, another aspect of the present invention provides a pharmaceutical formulation, preferably an oral pharmaceutical formulation, comprising particles prepared by drying water in which the compound of Formula 1 (preferably a compound having a crystalline form according to the present invention) is suspended and a hydrophilic polymer is dissolved.

[0021] In a preferred embodiment of the present invention, the hydrophilic polymer may be at least one selected from the group consisting of hypromellose, polyvinylpyrrolidone, polyvinyl alcohol, and hydroxypropyl cellulose, and among these, hypromellose is more preferred for the purposes of the present invention.

[0022] In order to achieve the object of the present invention, the content of such a hydrophilic polymer is preferably 30 to 80 wt % of the content of the compound of Chemical Formula 1, and more preferably 40 to 70 wt % of the content of the compound of Chemical Formula 1.

[0023] Another aspect of the present invention provides a pharmaceutical formulation, preferably an oral pharmaceutical formulation, comprising particles prepared by drying water in which a finely divided compound of Formula 1 (preferably a compound having a crystalline form according to the present invention) is suspended and a hydrophilic polymer is dissolved. The type and content of the hydrophilic polymer are the same as those described above.

[0024] Preferably, the micronized compound of Formula 1 has a d(50) of 5 μm or less, more preferably a d(50) of 2 μm or less.

[0025] Such a finely divided compound of Formula 1 can be obtained by using jet milling, high-pressure particle size grinding machines (microfluidics), etc., but the present invention is not limited to such a specific means of micronization.

[0026] The specific preparation method using the hydrophilic polymer of the present invention dramatically increased the bioavailability of the compound of Chemical Formula 1, and in particular, the bioavailability could be further increased by using a finely divided compound of Chemical Formula 1 (preferably a compound having a crystalline form according to the present invention). In the pharmaceutical formulation of the present invention, the hydrophilic polymer is expected to exhibit a thinner effect by playing various roles, such as stabilizing the compound of Chemical Formula 1, which is dispersed in a small size in suspension, to prevent further aggregation and increase in size, particularly in relation to the finely divided compound, and by binding to the surface of the compound of Chemical Formula 1, allowing it to be easily wetted by an aqueous solution. However, the present invention is not limited to such theoretical predictions. In particular, the effects of the present invention are closely related to the preparation method, as it is difficult to achieve the desired bioavailability by simply mixing the components.

[0027] In a preferred embodiment of the present invention, the process of pulverizing the compound of Chemical Formula 1 is carried out in water in which the compound of Chemical Formula 1 (preferably a compound having a crystalline form according to the present invention) is suspended and a hydrophilic polymer is dissolved. That is, in a preferred embodiment of the present invention, the process of pulverizing the compound of Chemical Formula 1 is carried out after mixing it with the hydrophilic polymer.

[0028] Another aspect of the present invention provides a pharmaceutical formulation, preferably an oral pharmaceutical formulation, comprising particles prepared by drying water in which a compound of Formula 1 (preferably a compound having a crystalline form according to the present invention, micronized) is suspended and a hydrophilic polymer and a surfactant are dissolved. The type and content of the hydrophilic polymer and the micronization techniques are the same as those described above.

[0029] The surfactant is preferably at least one selected from the group consisting of lauryl sulfate, poloxamer, glycerol monostearate, polyoxyethylene, and docusate salt, and for the purposes of the present invention, sodium lauryl sulfate is particularly preferred.

[0030] Such surfactants are preferably contained in the pharmaceutical preparation in an amount of 0.2 to 2% by weight based on the total weight of the pharmaceutical preparation.

[0031] The compound of Formula 1 according to the present invention (preferably a compound having a crystalline form according to the present invention, micronized) is suspended in water, in which a hydrophilic polymer and optionally a surfactant are dissolved, and the resulting particles are dried. The particles can be mixed with other carriers (e.g., excipients), lubricants, etc., and then filled into empty capsules to form a capsule formulation, or can be mixed with other carriers, lubricants, etc., and then compressed into tablets. In a preferred embodiment of the present invention, the pharmaceutical formulation is a tablet.

[0032] The compound of Formula 1 according to the present invention (preferably a compound having a crystalline form according to the present invention, which is micronized) is suspended in water, and a hydrophilic polymer and optionally a surfactant are dissolved in the water. The water can be sprayed onto another carrier and then dried to form particles.

[0033] Carriers, e.g., excipients, used in the present invention include microcrystalline cellulose, lactose, lactose hydrate, anhydrous lactose, mannitol, starch, dihydrogenated calcium phosphate, anhydrous dibasic calcium phosphate, etc. In a preferred embodiment of the present invention, the carrier is microcrystalline cellulose, lactose hydrate, mannitol, or a mixture thereof. In a preferred embodiment of the present invention, the carrier is microcrystalline cellulose and lactose hydrate. In a preferred embodiment of the present invention, the carrier is microcrystalline cellulose, lactose hydrate, and mannitol.

[0034] The pharmaceutical formulation of the present invention may further comprise a disintegrant, such as sodium starch glycolate, crospovidone, croscarmellose sodium, maltodextrin, or a mixture thereof. In a preferred embodiment of the present invention, the disintegrant is crospovidone.

[0035] The pharmaceutical formulation of the present invention may optionally further contain a lubricant. Lubricants are used to ensure smooth flow during capsule filling, packaging, etc., or to prevent phenomena such as sticking and capping during tablet compression. Suitable lubricants include colloidal silica, talc, magnesium stearate, or a mixture thereof. In a preferred embodiment of the present invention, the lubricant is magnesium stearate. [Effects of the Invention]

[0036] One aspect of the present invention provides a stable novel crystalline form of the compound of Chemical Formula 1 and a method for obtaining the crystalline form. Another aspect of the present invention provides a pharmaceutical formulation, preferably an oral pharmaceutical formulation, which contains the compound of Chemical Formula 1 (preferably the crystalline form of the compound according to the present invention) as an active ingredient and has ensured stability, high dissolution rate, high bioavailability, etc., and a method for producing the same. [Brief explanation of the drawings]

[0037] [Figure 1]1 shows the results of powder X-ray diffraction pattern (PXRD) evaluation of the crystalline substance prepared in Example 1. [Figure 2] 1 shows the results of measuring the absorbance of the crystalline material prepared in Example 1 by infrared spectroscopy (IR). [Figure 3] 1 is a graph comparing the blood concentrations obtained after oral administration of tablets produced in Production Example 10, which is an embodiment of the present invention, to beagle dogs with those of a comparative example. DETAILED DESCRIPTION OF THE INVENTION

[0038] The present invention will be described in detail below with reference to examples. However, the examples according to the present invention can be modified in various other forms, and the scope of the present invention is not limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to those skilled in the art.

[0039] Example 1: Preparation of a crystalline solid (crystalline form) of the compound of formula 1 4.14 kg of the solid compound of Formula 1 was dissolved in 17 kg of ethyl acetate and stirred at room temperature, while 15.5 kg of t-butyl methyl ether was gradually added. The mixture was stirred at 25°C for 3 hours to produce a homogeneous suspension, which was filtered and washed with 2.5 kg of t-butyl methyl ether. The mixture was dried in vacuo at 45°C for 15 hours to obtain 3.52 kg of a milky white crystalline solid (yield 85%).

[0040] 1H NMR (400MHz, DMSO-d6) δ8.64(d,J=4.8Hz,1H), 8.20(d,J=7.8Hz,1H), 7.68(dd,J=2.2,12.4Hz,1H), 7.65(dd,J=2.2,8. 5Hz,1H), 7.25(t,J=8.5Hz,1H), 7.16(d,J=4.8Hz,1H), 4.93-4.88(m,1H), 4.56-4.52(m,1H), 4.49(s,1H), 3.96-3.93(m ,1H), 3.67-3.63(m,1H), 3.59-3.56(m,1H), 3.41-3.37(m,1H), 3.20(s,3H), 3.14-3.11(m,1H), 3.01-2.98(m,1H), 2.10 -2.05(m,3H), 1.78-1.72(m,4H), 1.68-1.63(m,4H), 1.49-1.44(m,2H), 1.29(d,J=6.7Hz,3H);MS(FAB)m / z:544[M+H]+.

[0041] In order to obtain a suitable crystalline form, in addition to the ethyl acetate / t-butyl methyl ether combination conditions of Example 1, various other combinations were tried (e.g., using ethyl ether, isobutyl acetate, isopropyl acetate, methyl isobutyl ketone, n-butyl acetate, n-heptane, n-pentane, n-hexane, cyclohexane, methylcyclohexane, tetrahydrofuran, acetone, etc. instead of t-butyl methyl ether), but no crystalline form with satisfactory stability was obtained.

[0042] On the other hand, we fixed t-butyl methyl ether and tried solvents such as acetonitrile, dichloromethane, 1,2-dichloroethane, 1,2-dimethoxyethane, N,N-dimethylacetamide, 1,4-dioxane, methanol, ethanol, ethylene glycol, 1-butanol, 2-butanol, 1-propanol, 2-propanol, and propyl acetate instead of ethyl acetate, but we still could not obtain a crystal form stable enough for use as a pharmaceutical raw material.

[0043] In addition, we tried to prepare crystalline forms by changing all the combinations of ethyl acetate / t-butyl methyl ether and various combinations of solubilizing solvents and antisolvents, but we were unable to obtain the desired crystalline form.

[0044] When producing crystalline forms of pharmaceutical raw materials, selecting an appropriate solvent or solvent mixture is extremely important. The resulting crystalline solid must have excellent stability and desirable physical properties, and after drying, it must meet the residual solvent acceptance criteria of the ICH Guideline for Residual Solvents Q3C(R8). For these reasons, solvates (excluding hydrates) often cannot be used as pharmaceutical raw materials, even if they actually have excellent crystallinity.

[0045] The inventors have previously attempted to precipitate a solid by dissolving the compound of Formula 1 in a solubilizing solvent as a single solvent and then using a solvent evaporation method, but the resulting crystalline form was either a solvate or the residual solvent was not sufficiently removed even after prolonged drying under reduced pressure, resulting in a crystal form that significantly exceeded the ICH standard. In methods that induce solid precipitation using various solubilizing solvents and antisolvents, the resulting product was primarily amorphous or a solvate, or had other crystalline forms that were difficult to remove the residual solvent and therefore unusable as a pharmaceutical raw material, and even these products often had poor yields or discolored during the manufacturing process.

[0046] Therefore, the present inventors have confirmed that the combination of ethyl acetate / t-butyl methyl ether is suitable as a mixed solvent for preparing the crystalline form of Formula 1, which has properties suitable for use as a pharmaceutical raw material. The crystalline solid obtained in this manner not only has excellent stability and physical properties, but is also an anhydrous crystalline form, and analysis of the residual solvents revealed that the residual solvents do not exceed the respective allowable residual limits set forth in the ICH guidelines, making it suitable for use as a pharmaceutical raw material.

[0047] Experimental Example 1: Powder X-ray Diffraction (PXRD) Analysis of the Crystalline Form of Example 1 Powder X-ray diffraction patterns are an intrinsic characteristic of crystalline forms and are widely used to distinguish between polymorphisms and hydrates.

[0048] Crystallinity analysis of the crystalline compound prepared in Example 1 was performed using an X-ray powder diffractometer with Cu-Kα radiation. The measurement apparatus was equipped with reactive power, and the current was set at 45 kV and 40 mA. The divergence and scattering slits were set at 1°, and the receiving slit was set at 0.2 mm. A θ-2θ continuous scan was used from 5° to 35° 2θ at 3° / min (0.4 sec / 0.02° interval).

[0049] The experimental results are shown in Figure 1. As shown in Figure 1, the crystalline material prepared in Example 1 exhibited a powder X-ray diffraction pattern having peaks at 2θ diffraction angles of 5.3±0.2°, 7.5±0.2°, 10.1±0.2°, 10.6±0.2°, 11.5±0.2°, 14.3±0.2°, 14.7±0.2°, 15.0±0.2°, 15.5±0.2°, 16.0±0.2°, 16.3±0.2°, 16.5±0.2°, 19.1±0.2°, 19.3±0.2°, 19.7±0.2°, 20.3±0.2°, 20.6±0.2°, 21.4±0.2°, 21.8±0.2°, 22.6±0.2°, and 27.5±0.2°.

[0050] Experimental Example 2: Infrared Spectroscopy (IR) Analysis of the Crystalline Form of Example 1 The vibrational energy between atoms that make up the molecules of a material in a crystalline solid varies depending on the crystal structure, which means that different crystal forms exhibit different infrared absorbances. Therefore, the infrared absorbance of a particular crystalline material can be said to reflect the unique characteristics of that crystal form.

[0051] The infrared absorbance of the crystalline form prepared in Example 1 was analyzed using a PerkinElmer Fourier transform infrared spectroscopy (FT-IR), and the results are shown in Figure 2. As shown in Figure 2, -1 (aromatic CF), 1518cm -1(NH), 1579cm -1 (aromatic C=C), 1680cm -1 (C=O), 2911 cm -1 (aliphatic CH) and 3445 cm -1 The absorption pattern of (OH) was shown.

[0052] Experimental Example 3: Evaluation of the stability of the crystalline form of Example 1 The compound of Chemical Formula 1 is a raw material used in pharmaceutical applications, and it is necessary to ensure appropriate specifications and stability in order to determine the storage method and usage period of the pharmaceutical. In other words, the stability of pharmaceutical raw materials is one of the very important factors for maintaining the quality of the final product.

[0053] The stability of the crystalline solid of the compound of Formula 1 prepared in Example 1 was evaluated. Stability tests were conducted under long-term storage conditions (25±2°C, 60±5% RH) and accelerated storage conditions (40±2°C, 75±5% RH) in accordance with the ICH guidelines. Analysis was performed using HPLC to evaluate changes in content and the occurrence of related substances, and the results are shown in Table 1 below.

[0054] [Table 1]

[0055] As can be seen from the results in Table 1, the crystalline solid compound of Formula 1 of the present invention has excellent stability since there is almost no increase in related substances and almost no change in content when stored under long-term and accelerated storage conditions for up to 12 months and 6 months, respectively.

[0056] In addition, the crystalline form of samples stored for 3, 6, and 12 months (long-term storage test) of the stability test was measured using the same method as in Experimental Example 1, and it was confirmed that the crystalline form did not change. For BCS Class 2 substances such as the compound of Formula 1, it is very important that the crystalline form is stably maintained under storage conditions. The more poorly soluble the substance, the greater the impact of changes in crystalline form on the performance of the finished drug, causing changes in hygroscopicity, stability, dissolution, etc. Sometimes, even partial crystalline transition, in which only part of the drug substance changes rather than the entire substance, can significantly degrade the quality of the finished drug. The crystalline form of the present invention is suitable for use as a drug substance because it does not undergo polymorphic transition over time.

[0057] Finished Pharmaceutical Manufacturing Examples 1-3: Manufacturing of formulations with different particle sizes of the compound of Formula 1 Preparations 1 to 3, which are compositions containing the crystalline form of Example 1, were prepared according to Table 2 below. In Preparations 2 and 3, the compositions were exactly the same, and only the preparation methods were changed to produce formulations with different particle sizes.

[0058] [Table 2]

[0059] First, among the compositions in Table 2, in Production Example 1, the compound of Chemical Formula 1, which is the main component, was not subjected to any treatment.

[0060] In Preparation Example 2, materials used in the binding liquid were mixed and then homogenized using a homogenizer to prepare a binding liquid containing particles of the compound of Formula 1.

[0061] In Preparation Example 3, materials used in the binding liquid were mixed and then pulverized under high-energy milling conditions using a high-pressure particle size pulverizer (Microfluidics) to prepare a binding liquid containing finely pulverized particles of the compound of Chemical Formula 1.

[0062] In Preparation Examples 2 and 3, the particle size of the compound of Chemical Formula 1 was measured in the binder solution state, i.e., in the state where SLS and HPMC were mixed. The binder solution was in a suspension state, with SLS and HPMC dissolved in purified water, and only the main component was suspended in a solid state, making it possible to measure the particle size of the main component. The particle size of the compound of Chemical Formula 1 was measured using a laser scattering particle size analyzer (Mastersizer 2000, manufacturer: Malvern Instruments). The compound of Chemical Formula 1 particles prepared in Preparation Examples 1 to 3 and the solvent (purified water) were added to a beaker (1,000 ml) and stirred at 1,500 rpm, and the homogeneously suspended dispersed phase particles were measured. The results are shown in Table 3.

[0063] <Particle size measurement conditions> Equipment:Malvern Mastersizer 2000 / Hydro 2000MU Sample amount: 0.5g Sample refractive index: 1.520 Interpretation model: General purpose Sensitivity:Normal Sample measurement time: 10 seconds Interpretation range: 0.020~2000.0μm

[0064] [Table 3]

[0065] In Preparation Examples 2 and 3, granules were produced using a fluid bed granulator. A binder solution was sprayed onto microcrystalline cellulose and lactose hydrate, which were used as carriers, and granulated under conditions of an injection temperature of 70-85°C, an internal temperature of 35-45°C, and an injection rate of 6-30 g / min. The main ingredients or granules produced in Preparation Examples 1 to 3 were filled into capsules.

[0066] Experimental Example 4: Effect of particle size on dissolution rate The preparations of Production Examples 1 to 3 were tested by the second dissolution test method (paddle method) of the Korean Pharmacopoeia (Korean Pharmaceutical Codex). The dissolution medium was purified water, and the dissolution test was carried out at a paddle speed of 50 rpm and 37°C, and quantified by HPLC (manufacturer: Agilent) under the following conditions. The results are shown in Table 4.

[0067] <HPLC measurement conditions> Detector: Ultraviolet absorption photometer (measurement wavelength: 254 nm) Column: Inertsil ODS-4 C18 (4.6×150 mm, 5 μm) Flow rate: 1.0 mL / min Column temperature: 35°C Mobile phase: Phosphoric acid solution with pH 3.0: Acetonitrile = 4:6 (v / v%) Injection volume: 20 μL

[0068]

Table 4

[0069] As can be seen from Table 4 above, the dissolution rate and dissolution percentage were affected by the particle size, and it was confirmed that the smaller the particle size, the higher the dissolution rate. This is presumably because the smaller the particle size, the larger the surface area, but the present invention is not limited to such a theoretical mechanism. The dissolution percentage at the 45-minute point increased by about two times in Production Examples 2 to 3 with a decreased particle size compared to Production Example 1. Furthermore, it was determined that the use of a hydrophilic polymer was further required for improving the dissolution percentage, and an evaluation thereof was conducted.

[0070] Manufacturing Examples 4 to 6: Changes in dissolution depending on the hydrophilic polymer content In order to comparatively evaluate the influence of the content of the hydrophilic polymer on dissolution, tablets of Production Examples 4 to 6 were produced according to the composition shown in Table 5 below. In Production Example 4, the hydrophilic polymer was used only in Binding Solution 1, and in Production Examples 5 to 6, Binding Solution 2 in which the hydrophilic polymer was dissolved was further used.

[0071]

Table 5

[0072] In Preparation Example 4, granules were prepared by the method of Preparation Example 3, and then the prepared granules were mixed with mannitol, crospovidone, and magnesium stearate, and then formed into tablets.

[0073] In Preparation Examples 5 and 6, binding solution 1 was prepared by the method of Preparation Example 3, hypromellose and purified water were mixed to prepare binding solution 2, and binding solution 1 and binding solution 2 were mixed to prepare the final binding solution. Then, microcrystalline cellulose and lactose hydrate were used as carriers to prepare granules by the method of Preparation Example 3. The prepared granules were mixed with mannitol, crospovidone, and magnesium stearate and then formed into tablets.

[0074] The particle sizes of the compounds of Chemical Formula 1 prepared in Preparation Examples 4 to 6 were measured using a laser scattering particle size analyzer (Mastersizer 2000, manufactured by Malvern Instruments), and the results are shown in Table 6 below.

[0075] [Table 6]

[0076] Experimental Example 5: Changes in dissolution rate due to hydrophilic polymer content As in Experimental Example 4, the tablets of Preparation Examples 4 to 6 were tested according to Method 2 (paddle method) of the Korean Pharmacopoeia. The dissolution test was carried out using purified water at a paddle speed of 50 rpm and 37°C. Quantitative analysis was carried out by HPLC (manufacturer: Agilent) under the same conditions as in Experimental Example 4. The results are shown in Table 7.

[0077] [Table 7]

[0078] As can be seen from Table 7, in the case of micronized particles of the compound of Chemical Formula 1, the dissolution rate of Preparation Example 4, in which the amount of hydrophilic polymer (hypromellose) was 15 mg per tablet (15 parts by weight per 100 parts by weight of the compound of Chemical Formula 1), hardly increased. However, in Preparation Examples 5 and 6, in which the amount of hydrophilic polymer was 50 mg to 60 mg per tablet (50 to 60 parts by weight per 100 parts by weight of the compound of Chemical Formula 1), the dissolution rate increased by about 5 to 6 times compared to Preparation Example 4.

[0079] Production Examples 7 to 10: Changes in dissolution depending on the amount of surfactant The effect of the amount of surfactant on dissolution was compared and evaluated. Tablets of Production Examples 7 to 10 were produced according to the compositions shown in Table 8 below.

[0080] [Table 8]

[0081] Preparation Examples 7 to 10 were prepared by the method of Preparation Example 6. The amount of surfactant used per tablet was 1.5 to 8 mg, and the differences due to the amount of surfactant and the preparation method were evaluated using binding solution 1 and binding solution 2, respectively.

[0082] The particle sizes of the compounds of Chemical Formula 1 prepared in Preparation Examples 7 to 10 were measured using a laser scattering particle size analyzer (Mastersizer 2000, manufactured by Malvern Instruments), and the results are shown in Table 9 below.

[0083] [Table 9]

[0084] Experimental Example 6: Changes in dissolution rate depending on surfactant content The tablets of Preparation Examples 7 to 10 were tested using Method 2 (paddle method) of the Korean Pharmacopoeia dissolution test. The dissolution medium was purified water (with 0.3 wt% sodium lauryl sulfate (SLS) added), and the dissolution test was carried out at a paddle speed of 50 rpm and 37°C. Quantitative analysis was carried out using HPLC (manufacturer: Agilent) under the same conditions as the above-mentioned method. The results are shown in Table 10 below.

[0085]

Table 10

[0086] As shown in Table 10 above, despite the amount of surfactant varying from 1.5 mg to 8 mg per tablet, the final dissolution rate reaching 60 minutes was similar.

[0087] Experimental Example 7: Stability test of formulation After storing Production Example 10 under accelerated conditions (40 ± 2°C, 75% RH ± 5%) and long-term conditions (25 ± 2°C, 60% RH ± 5%), the content and related substances were evaluated. The results are shown in Tables 12 and 13. The unit is expressed as %.

[0088] <HPLC measurement conditions_content> Detector: Ultraviolet absorption photometer (measurement wavelength: 254 nm) Column: Inertsil ODS-4 C18 (4.6 × 150 mm, 5 μm) Flow rate: 1.0 mL / min Column temperature: 35°C Mobile phase: pH 3.0 phosphate solution: acetonitrile = 4:6 (v / v%) Injection volume: 20 μL Measurement range: 7 minutes <HPLC measurement conditions_purity> Detector: Ultraviolet absorption photometer (measurement wavelength: 254 nm) Column: YMC Hydrosphere C18 5 μm, 4.6 × 250 mm Flow rate: 1.0 mL / min Column temperature: 35°C Mobile phase A: Dissolve 0.68 g of potassium dihydrogen phosphate in water to make 1,000 mL, and adjust the pH to 3.0 with phosphoric acid.

[0089] Mobile phase B: Mix acetonitrile and methanol in a ratio of 95:5.

[0090] Measurement range: 60 minutes Gradient elution conditions:

[0091] [Table 11]

[0092] [Table 12]

[0093] [Table 13]

[0094] As shown in Tables 12 and 13, the results of the content evaluation and purity test during the stability test period confirmed that the formulation of the present invention was stable without any significant changes.

[0095] Experimental Example 8: Comparative pharmacokinetic study of formulations Tablets containing 25, 50, and 100 mg of the crystalline solid of Formula 1 were prepared using the conditions of Preparation Example 10. The tablets were orally administered to beagle dogs, and an experiment was conducted to analyze their pharmacokinetic properties.

[0096] The compound of Chemical Formula 1 was orally administered to Beagle dogs at a single dose of 25, 50, and 100 mg tablets / head, a single dose of 25 mg liquid / head, and a single intravenous dose of 2 mg / kg. Blood samples were taken before administration of the test substance (0), and 15 and 30 minutes, and 1, 1.5, 2, 4, 6, 8, 10, 16, and 24 hours after administration, and the blood concentrations of the compound of Chemical Formula 1 were measured.

[0097] Liquid / head: A single oral administration preparation was prepared at 1.25 mg / ml by homogenously suspending 275 mg of the compound of Formula 1 in 220 ml of a pre-prepared 2.5% HP-βCD (sterile water solution). A single intravenous administration preparation was prepared at 2 mg / ml by dissolving 300 mg of the compound of Formula 1 in 150 ml of a pre-prepared 10% HP-βCD (sterile water solution).

[0098] As a result of observing the general symptoms after administration, no abnormal symptoms due to the administration of the test substance were observed during the entire experimental period.

[0099] Validation of the analytical method in canine plasma samples showed no influence of interfering substances and good linearity in the concentration range of 5 to 5,000 ng / mL (plasma), as well as consistent results for intra- and inter-assay accuracy and precision. The method demonstrated sufficient selectivity, linearity, accuracy, and precision for application to sample analysis.

[0100] The drug concentration-time curves analyzed from plasma obtained after administration of the compound of Formula 1 in tablet and liquid form and the pharmacokinetic parameters calculated using the BA Calc 2007 program showed the profiles shown in Figure 3 and Table 14.

[0101] [Table 14]

[0102] As shown in Table 14, in the case of tablets, the absorption rate of the drug at the time of administration was AUC last The time to maximum drug absorption, T max indicates a range of 1 to 2 hours at each dose, and the average half-life (t 1 / 2 ) showed 4.42 hours, 5.29 hours, and 6.89 hours, indicating that the excretion rate slowed as the administered volume increased.

[0103] When comparing the 25 mg / head tablet group and the liquid group, the mean absorption AUC last In the case of the compound of formula 1, the oral formulation was optimized in terms of solubilization and bioabsorption, demonstrating that the oral formulation of the compound of formula 1, a poorly soluble drug, achieved 96.7% of the solubilization rate of the tablet group compared to the liquid group.

[0104] As a result, the present invention provides a stable novel crystalline solid of the compound of Chemical Formula 1, and by using the same, an oral formulation with excellent stability and excellent oral absorption rate is secured.

Claims

1. A crystal of the compound of [Chemical Formula 1] below, having a powder X-ray diffraction pattern with peaks at 2θ diffraction angles of 5.3±0.2°, 7.5±0.2°, 10.1±0.2°, 10.6±0.2°, 11.5±0.2°, 14.3±0.2°, 14.7±0.2°, 15.0±0.2°, 15.5±0.2°, 16.0±0.2°, 16.3±0.2°, 16.5±0.2°, 19.1±0.2°, 19.3±0.2°, 19.7±0.2°, 20.3±0.2°, 20.6±0.2°, 21.4±0.2°, 21.8±0.2°, 22.6±0.2°, and 27.5±0.2° in powder X-ray diffraction (PXRD) analysis. [Chemical formula 1]

2. (S1) dissolving a compound of the following formula 1 in ethyl acetate; [Chemical formula 1] (S2) adding t-butyl methyl ether to the ethyl acetate solution of step S1 to crystallize the solution; (S3) filtering the suspension from step S2 to obtain a crystalline compound of formula 1.

3. A pharmaceutical formulation comprising a compound of [Chemical Formula 1], wherein the compound of [Chemical Formula 1] has the crystal described in claim 1.

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