Mesylate of triazolopyrazine derivative

Abstract

It relates to a salt of methanesulfonic acid and a triazolopyrazine derivative of Chemical Formula 1 (mesylate), a pharmaceutical composition containing the salt, a method for producing the salt, and a therapeutic use of the salt. [Chemical Formula 1] JPEG2025520579000022.jpg54102

Description

Technical Field

[0001] Triazolopyrazine derivatives are, as is known, substances used in treating hyperproliferative diseases by suppressing the activity of c-Met kinase. However, in the prior literature, many free base forms, which are not the salt forms of such triazolopyrazine derivatives, are presented.

[0002] A salt form is required to improve the pharmacological usefulness of the free base form of the triazolopyrazine derivative. Therefore, in the present invention, it is desired to provide a mesylate salt that maintains the pharmacokinetics and pharmacological activity of the free base form and improves useful physical properties, and a method for producing the same.

Background Art

[0003] The present invention relates to a novel mesylate salt compound of a triazolopyrazine derivative represented by the following Chemical Formula 1, a method for producing the same, and a pharmaceutical composition containing the same.

[0004]

Chem.

[0005] The inventors of the present invention have earnestly studied and made efforts to develop a composition for efficiently preventing or treating various hyperproliferative disorders induced by abnormal tyrosine kinase activity by discovering the compound having inhibitory activity against tyrosine kinase. As a result, the inventors have found that a novel salt compound of the compound of Chemical Formula 1, which has not been known heretofore, improves the storage stability and photo stability of the compound while maintaining the function of suppressing the activity of the c-Met kinase of the compound, and thus completed the present invention.

[0006] According to the present invention, the novel mesylate compound of the compound of Chemical Formula 1 of the present invention binds to hepatocyte growth factor (HGF) and activates phosphorylation, thereby promoting cell proliferation, migration, and neovascularization, and significantly suppressing the activity of c-Met kinase.

[0007] Therefore, the salt compound of the present invention can be usefully utilized for the treatment or prevention of various hyperproliferative diseases mediated by excessive angiogenesis due to abnormal cell proliferation activation.

[0008] In addition, the novel mesylate compound of the compound of Chemical Formula 1 exhibits physical properties different from those of the pure compound, such as melting point, hygroscopicity, solubility, flow characteristics, or thermodynamic safety.

[0009] Thereby, the salt compound of the compound of Chemical Formula 1 provided by the present invention can be used to select the most suitable form for pharmacological use in the drug manufacturing process or in the manufacturing process of separate drug dosage forms such as purification, capsules, ointments, suspensions, etc., or in the manufacturing of drug forms having optimal pharmacokinetic properties.

[0010] According to another aspect of the present invention, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the novel mesylate compound of Chemical Formula 1. For example, the present invention provides a pharmaceutical composition for the prevention or treatment of a hyperproliferative disorder containing the novel salt compound of the compound of Chemical Formula 1 described above as an active ingredient.

[0011] As used herein, the term "abnormal proliferative disease" refers to a pathological condition resulting from excessive cell growth, division, and migration that is not controlled by the general regulatory means in a normally growing animal body. The abnormal proliferative diseases that can be prevented or treated with the compositions of the present invention include, for example, cancer, diabetic retinopathy, retinopathy of prematurity, corneal transplant rejection, neovascular glaucoma, iritis, proliferative retinopathy, psoriasis, rheumatoid arthritis, osteoarthritis, autoimmune diseases, Crohn's disease, restenosis, atherosclerosis, intestinal adhesions, ulcers, hepatic disorders, glomerulonephritis, diabetic kidney disease, malignant nephrosclerosis, thrombotic microangiopathy, tracheal transplant rejection, and renal glomerular diseases, but are not limited thereto, and include all abnormal proliferative diseases caused by abnormal cell proliferation and excessive formation of new blood vessels.

[0012] More preferably, one of the cancers of abnormal proliferative diseases that can be prevented and treated with the compositions of the present invention is lung cancer, gastric cancer, pancreatic cancer, colorectal cancer, ovarian cancer, kidney cancer, prostate cancer, or brain tumor.

[0013] When the composition of the present invention is manufactured as a pharmaceutical composition, the pharmaceutical composition of the present invention contains a pharmaceutically acceptable carrier. The pharmaceutically acceptable carriers contained in the pharmaceutical composition of the present invention are those commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, etc., but are not limited thereto. The pharmaceutical composition of the present invention can further contain lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. in addition to the above components.

[0014] The pure substance of the triazolopyrazine derivative represented by the above Chemical Formula 1 often has aspects where physical properties such as solubility and thermal stability are not suitable for manufacturing general pharmaceutical compositions.

[0015] Therefore, the inventors of the present invention conducted various salt screenings to develop a salt compound suitable for producing a pharmaceutical dosage form such as improving the solubility and safety of the triazolopyrazine derivative, and through this, a novel maleate compound with high solubility and excellent safety while continuously maintaining the pharmacological activity of the compound was developed.

Prior Art Documents

Patent Documents

[0016]

Patent Document 1

Summary of the Invention

[0017] In order to improve the problems that the storage stability and photo-stability of the pure substance of the triazolopyrazine derivative represented by the above Chemical Formula 1 are low and the processability is not good in pharmaceutical manufacturing, salt screening was carried out on the above compound, and by developing a novel mesylate compound of the above compound, the above problems could be solved.

Means for Solving the Problems

[0018] The present invention was able to improve the storage stability and photo-stability of the triazolopyrazine derivative (ABN401) represented by the above Chemical Formula 1 by developing a novel mesylate (Mesylate, Methanesulfonic acid) compound of the above triazolopyrazine derivative.

[0019] In addition, the present invention provides a method for producing the above mesylate compound and a pharmaceutical composition containing the above salt compound as an active ingredient.

Effects of the Invention

[0020] The present invention relates to a novel mesylate compound of a triazolopyrazine derivative represented by the above Chemical Formula 1 and a method for producing the same. The novel mesylate compound provided by the present invention improves storage stability and photo stability, which were problems of pure substances, and improves the pharmaceutical manufacturability and processability of the compound.

[0021] By efficiently suppressing the activity of c-Met tyrosine kinase, the mesylate compound can be usefully utilized as a therapeutic agent for various abnormal proliferative diseases related to excessive cell proliferation and growth caused by abnormal kinase activity, such as cancer, psoriasis, rheumatoid arthritis, diabetic retinopathy, and the like. In addition, a pharmaceutical composition for suppressing c-Met tyrosine kinase activity containing the novel salt compound as an active ingredient and a pharmaceutical composition for preventing or treating a hyper proliferative disorder are provided.

Brief Description of the Drawings

[0022]

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[0023] The mesylate, which is a novel salt of the compound, can be usefully utilized as a therapeutic agent for various abnormal proliferative diseases related to excessive cell proliferation and growth due to abnormal kinase activity, such as cancer, psoriasis, rheumatoid arthritis, diabetic retinopathy, etc., by efficiently suppressing the activity of c-Met tyrosine kinase. Further provided are a pharmaceutical composition for suppressing c-Met tyrosine kinase activity containing the novel salt compound as an active ingredient and a pharmaceutical composition for preventing or treating a hyper proliferative disorder.

[0024] On one aspect, the mesylate is an enantiomer of the triazolopyrazine derivative represented by Chemical Formula 1. Desirably, the triazolopyrazine derivative represented by Chemical Formula 1 is optically pure. On another aspect, the optical purity is at least 95%, at least 97%, at least 99% or essentially 100%.

[0025] On one aspect, the mesylate is formed in any one of the salt forms of Mes1, Mes2 or Mes3. Probably, the mesylate is likely to exist in the form of Mes2. On one aspect, the mesylate is physically stable at a temperature of 20 - 50 °C and a relative humidity of 35 - 80% for at least 2 days. On one aspect, the mesylate is physically stable for 24 months under ambient environmental conditions. Also, the mesylate has a purity of the active ingredient of at least 95%, and it is desirable that the purity is about 100%.

[0026] On one side, the mesylate is composed of characteristic peaks at 15.5 - 16.0 2θ (deg), 17.5 - 18.0 2θ (deg) and 21.5 - 22.0 2θ (deg) in a high-efficiency X-ray diffraction grating (HT-XRPD) pattern. Desirably, the HT-XRPD of the mesylate has characteristic peaks substantially corresponding to those below.

[0027]

Table 1

[0028]

Table 2

[0029] On one side, the present invention provides a method for producing the mesylate. The production method includes: a) a step of adding the compound of Chemical Formula 1 to a reactor containing a solvent; b) a step of stirring the solvent and the compound in the reactor; c) a step of adding methanesulfonic acid to the solution adjusted in step b); and d) a step of cooling the solution adjusted in step c to precipitate the mesylate.

[0030] On one side, the solvent can be selected from acetonitrile, acetone, 1,2-dimethoxyethane, n-heptane, isopropyl alcohol, water, or tetrahydrofuran (THF). On one side, the methanesulfonic acid added to the solution adjusted in step b) is added in an equivalent amount of 1:1.5 to 1:2.5 with respect to the compound of Chemical Formula 1. Desirably, the equivalent ratio is 1:1.9 to 1:2.3 and the solvent is acetonitrile. On one side, step b) is carried out at about 45 to 55 °C for at least 1 hour.

[0031] On one side, the present invention provides a pharmaceutical composition containing the mesylate and a pharmaceutical carrier. The pharmaceutically acceptable carrier contained in the pharmaceutical composition is one or more components selected from the group including lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Further, the pharmaceutical composition provided by the present invention may further contain lubricants, wetting agents, sweeteners, flavoring agents, emulsifying agents, suspending agents, preservatives, etc. in addition to the above components.

[0032] In one aspect, the present invention provides a method for reducing the activity of c-Met kinase by administering to a patient in need of a pharmaceutical composition obtained by adding an effective amount of the mesylate salt. In one aspect, the patient is suffering from a disease associated with hyperproliferative activation of cells, and by way of example, the disease is lung cancer, visceral cancer, pancreatic cancer, rectal cancer, ovarian cancer, kidney cancer, prostate cancer or brain tumor. In one aspect, the method is a method for treating or preventing hyperproliferative activation of cells.

[0033] <General> [Table 3]

[0034] <Solvent> [Table 4]

[0035] <Counterion> [Table 5] The present invention will be described in detail by the following examples, but the technical scope of the present invention is not limited to those examples. [Examples]

[0036] The compound of Chemical Formula 1 (ABN-401) was supplied by Abion, and the other compounds were supplied by Fisher Scientific, Sigma Aldrich or VWR. The compounds used were of research grade, and the solvents used for LCMS analysis were of LCMS grade.

[0037] Determination of Solubility The solubility was determined by adding small aliquots of solvent (up to 50 μL for up to 1 mL and 100 μL for over 1 mL) to the solid starting material until complete dissolution or until a maximum volume of 4 mL was reached.

[0038] The solvent was then evaporated and the resulting solid was analyzed by HT-XRPD. The experimental results and detailed content are shown in Table 6.

[0039]

Table 6

[0040] Thermal stability The thermal stability experiment of ABN-401 was conducted to confirm the chemical stability of the starting material that dissolved when exposed to high temperatures.

[0041] The ABN-401 solution was adjusted to a concentration of about 0.3 mg / mL in a solvent consisting of acetone, 1,2-dimethoxyethane, and acetonitrile. The solution was exposed to room temperature for 1 hour and then two sample aliquots were sequentially heated at 50 °C and 80 °C for 1 hour each. These solutions were analyzed by LCMS.

[0042] Furthermore, the sample placed at room temperature for 24 hours was re-measured by LCMS to confirm chemical alteration over time.

[0043] No significant chemical alteration was confirmed in the samples held at 50 °C and 80 °C. The results are summarized in Table 7.

Table 7

[0044] Salt screening experiment The salt screening experiment was initiated by adjusting the concentration of the ABN-401 solution in acetone, 1,2-dimethoxyethane, and acetonitrile to approximately 25, 13, and 8 mg / ml, respectively.

[0045] Counterions were added to the API solution, and the content ratios were API:counterions of 1:1.1, 1:2.1, and 1:3.1, respectively. The experiment derived a temperature profile including three heating-cooling cycles between 5 - 50 °C and was deteriorated at 25 °C for 3 days (Figure 1).

[0046] After the deterioration time, since solid particles were separated from the liquid phase, they were analyzed through HT-XRPD after drying under reduced pressure at 50 °C.

[0047] The solution and the mother liquid phase were evaporated at room temperature, and the remaining solid particles were analyzed through HT-XRPD. Subsequently, all solid particles were exposed to accelerated deterioration conditions of 40 °C / 75%RH for 48 hours and re-measured by HT-XRPD to confirm their physical stability.

[0048] Scale-up of the salt The mesylate, Mes2, was scaled up on a 0.5 gram scale (Experiment ID: SSm71). 500 mg of the base form was dissolved in 23 ml of acetone at room temperature to a concentration of 26 mg / ml.

[0049] Methanesulfonic acid (1M in water) was added such that the molar ratio of API:CI was 1:2.1. With the addition of the counterion, precipitation of the salt was observed. The suspension was heated to 50 °C and continuously stirred at 50 °C for 8 hours. The salt suspension was cooled to 25 °C. The solid particles were separated and dried under reduced pressure at 50 °C 10 mbar and then analyzed by HT- / HR-XRPD, TGA / TGMS, DSC, LCMS, 1H-NMR, and DVS.

[0050] The camcylate Camp2 was scaled up to a 0.3 gram scale (Experiment ID: SSm73). 300 mg of the base form was dissolved in 15 ml of ACN to a concentration of 20 mg / ml. 136 mg of camphor-10-sulfonic acid was added such that the molar ratio of API:CI was 1:1.1. The counter ion was dissolved with continuous stirring at 50 °C for 1 hour, the solution was cooled to 25 °C and aged for 72 hours. Precipitation of the salt occurred during the cooling profile and after separating the solid precipitate and drying it under reduced pressure at 50 °C and 10 mbar, it was analyzed by HT- / HR-XRPD, TGA / TGMS, DSC, LCMS, 1H-NMR and DVS.

[0051] Physical and Chemical Stability Physical stability experiments for the mesylate (Mes2), camcylate (Camp2) and maleate (Mae2) were conducted at specific temperatures and relative humidities (25 °C / 60%RH and 40 °C / 75%RH) for 2 years.

[0052] Similarly, the above stability experiments were also carried out on the base form of ABN-401 as a reference substance.

[0053] At 1, 3, 15, and 24 months, XRPD, TGMS, and HPLC tests were performed on the vials containing the particles to confirm whether there were any special changes in the solid particles and the content of water and solvent, and also to confirm whether there was any deterioration of the compound in the storage state stored under other conditions.

[0054] Conditions for Experimenting the Stability of Camcylate (Camp2), Maleate (Mae2), Mesylate (Mes2) and the Base Form (FB) of ABN-401

Table 8

[0055] High-Efficiency X-Ray Powder Diffraction Analysis The HT-XRPD pattern was collected by a crystal T2 high-efficiency XRPD instrument.

[0056] The plate was mounted on a Bruker General Area Detector Diffraction System (GADDS), and the system was equipped with a VANTEC-500 gas area detector to correct for intensity and shape changes. The correction for accuracy with respect to peak position was performed using the NIST SRM1976 standard (Corundum).

[0057] Data collection was carried out using Cu Kα monochromatic light in the 2θ range between 1.5° and 41.5° at room temperature, and the method is the most unique for XRPD patterns. The diffraction pattern of each well was collected through an exposure of 90 seconds per frame in two 2θ ranges (1.5° ≤ 2θ ≤ 21.5° first frame, 19.5° ≤ 2θ ≤ 41.5° second frame).

[0058] No background removal or curve smoothing operations were performed on the XRPD patterns.

[0059] The carrier material used during HT-XRPD measurements was transparent to X-rays and had a very slight effect on the background.

[0060] High-Resolution X-Ray Powder Diffraction Analysis Powder data were collected using a D8 diffractometer, where Cu K1 (1.54056 Å) using germanium monochromatic light at room temperature was used.

[0061] The data were collected using a LynxEye detector for solids through 2θ steps of 0.016° in the 2θ range between 4° and 45° at a step speed of 22 seconds / step. The samples were measured using a glass capillary with an outer diameter of 0.3 mm and a length of 8 mm.

[0062] TGMS Analysis Mass loss due to the loss of water and solvent from the crystal was measured through TGA. The weight of the sample was monitored and heated in a TGA / DSC3+STARe system, resulting in a weight-temperature curve and a heat flow signal. The TGA / DSC3+ results were corrected for temperature using indium and aluminum samples. The sample (circa 2 mg) was placed in a 100 μL aluminum crucible and sealed. A fill hole was formed for sealing, and the crucible was heated from 25 °C to 300 °C at a heating rate of 10 °C / min during the TGA process. Dry nitrogen gas was used during the purging process.

[0063] The gas emitted from the TGA sample was analyzed by an Omnistar GSD301 T2 (Pfeiffer Vacuum GmbH, Germany) mass spectrometer. The analyzer is capable of mass spectrometry at temperatures in the range of 0 - 200 amu.

[0064] DSC Analysis DSC temperature recording was performed through a heat flux DSC3+STARe system. The DSC3+ was corrected for temperature and enthalpy using trace amounts of indium (m.p. = 156.6 °C; δHf = 28.45 J / g) and zinc (m.p. = 419.6 °C; δHf = 107.5 J / g).

[0065] The sample was sealed in a standard 40 μL aluminum pan with a pinhole opened and heated from 25 to 300 °C at a heating rate of 10 °C / min during the DSC process. Dry nitrogen gas with a flow rate of 50 mL / min was used during the purging process.

[0066] H-NMR Analysis The 1H-NMR spectrum was recorded at room temperature through a 500 MHz Varian instrument. The sample was dissolved in DMSO for analysis. The data was processed by an ACD Labs software spectrum processor.

[0067] LCMA Analysis Characterization of the starting material and purification of the novel crystalline form in the prebase form Method name: S19061_0.3M Sample: Concentration 0.3 mg / ml, Diluent: Acetonitrile

[0068]

Table 9

[0069] Assay and Purification of the Novel Crystal Form Salt Method Name: S19061_01.M Sample: Concentration 0.3 mg / ml, Diluent: Acetonitrile / Water (50 / 50)

[0070]

Table 10

[0071] The hygroscopicity was classified according to the European Pharmacopoeia hygroscopicity classification.

[0072] The classification of the moisture absorption rate at 25°C / 80%RH (24 hours) is as follows.

[0073] Mass change < 0.2% - Non-hygroscopic Mass change > 0.2% and < 2% - Slight hygroscopicity Mass change > 2% and < 15% - Moderate hygroscopicity Mass change > 15% - High hygroscopicity Mesylate Figure 4 shows the XRPD patterns of all mesylates with respect to the pre-base form (Form A).

[0074] Mes1 is a mesylate obtained from an experiment in which methanesulfonic acid was added so that the molar ratio of API:CI was 1:1 in three types of crystallization solvents.

[0075] Table 11 shows the experimental conditions for obtaining Mes1. The solid selected for the subsequent characterization process is a crystalline form salt formed when acetone and 1 molar equivalent of methanesulfonic acid were added (Experiment ID: SSm4). Mes1 showed physical stability when exposed to AAC (40 °C / 75% RH) for 2 days (Figure 6).

[0076] Experimental conditions for obtaining Mes1. * indicates the solid formed after evaporation of the solvent. [Table 11]

[0077] The TGA / TGMS analysis results for Mes1 showed a 4.3% mass loss due to acetone and water at 140 °C based on the MS signal. Thermal decomposition was observed at temperatures above 230 °C.

[0078] The heat flow signal of the TGA showed a broad endothermic reaction due to mass loss around approximately 160 °C, which is associated with the melting of the anhydrous form. The above thermal analysis results imply that Mes1 is a complex of a hydrated / dissolved crystalline form salt.

[0079] Mes2 is a mesylate obtained from an experiment in which methanesulfonic acid was added so that the molar ratio of API:CI was 1:2 in three types of crystallization solvents, and also added so that the ratio was 1:3 in ACN. Table 12 shows the experimental conditions for obtaining Mes12. The solid selected for the subsequent characterization process is a crystalline form salt formed when acetone and 2 molar equivalents of methanesulfonic acid were added (Experiment ID: SSm5). Mes2 showed physical stability when exposed to AAC (40 °C / 75% RH) for 2 days.

[0080] Experimental conditions for obtaining Mes2

Table 12

[0081] The TGA / TGMS analysis results for Mes2 showed a gradual mass loss of 4.6% between 25 and 150 °C due to water based on the MS signal (4.6% corresponds to 2.0 water molecules per salt molecule). Thermal decomposition was observed at 220 °C and above.

[0082] The heat flow signal of the TGA showed a broad endothermic reaction due to water loss and an endothermic reaction around about 210 °C, which is related to the melting of the anhydride. The thermal analysis results imply that Mes2 is the dihydrate of dimesylate.

[0083] The DSC analysis results of Mes2 showed a broad endothermic reaction from 25 to 120 °C, which may be related to water loss. A double endothermic reaction was recorded around about 218.2 °C, presumably due to the melting of the potential anhydrous salt, followed by an exothermic reaction estimated to be due to thermal decomposition at 223 °C.

[0084] The chemical purity of Mes2 was measured by LCMS and was 100%. This confirmed the presence of API in the solid phase of Mes2.

[0085] The 1H-NMR analysis results for Mes2 are presented in Figure 11. The shift of the API resonance visible in the spectrum of Mes2 implies that proton transfer occurred with the acidic counterion in the free base. The ratio of API:CI appeared as 1:2, and residual acetone was observed at 2.2 ppm.

[0086] Mes3 is a mesylate salt obtained from an experiment in which methanesulfonic acid was added in acetone and 1,2-dimethoxyethane solvents such that the molar ratio of API:CI was 1:3. The solid selected for subsequent characterization was a crystalline form salt formed when acetone and methanesulfonic acid were added in 3 molar equivalents (Experiment ID: SSm6). Table 13 shows the experimental conditions for obtaining Mes3. Although crystallinity was maintained, Mes3 showed solubility when exposed to AAC (40 °C / 75% RH) for 2 days.

[0087] Experimental conditions for obtaining Mes3

Table 13

[0088] The TGA / TGMS analysis results for Mes3 showed a 5.6% mass loss from 25 to 140 °C due to water based on the MS signal (5.6% corresponds to 2.8 water molecules per salt molecule). Thermal decomposition was observed at temperatures above 220 °C.

[0089] The heat flow signal of the TGA showed a broad endothermic reaction due to water loss around ~200 °C, followed by thermal decomposition. The above thermal analysis results imply that Mes3 is a hydrated crystalline form salt.

[0090] Camphorsulfonate Figure 14 compares the HT-XRPD patterns of the camphorsulfonate salt with the starting material (Form A).

[0091] Camp1 is a salt form obtained from an experiment in which camphor-10-sulfonic acid was added in acetone and 1,2-dimethoxyethane solvents such that the molar ratio of API:CI was 1:1. Table 14 shows the experimental conditions for obtaining Camp1. The solid selected for subsequent characterization was a crystalline form salt formed when acetone and camphor-10-sulfonic acid were added in 1 molar equivalent (Experiment ID: SSm14). Camp1 was physically stable when exposed to AAC (40 °C / 75% RH) for 2 days.

[0092] Experimental conditions for obtaining Camp1

Table 14

[0093] The TGA / TGMS analysis results for Camp1 showed a 3.3% mass loss between 25 and 150 °C due to water based on the MS signal (3.3% corresponds to 1.5 water molecules per salt molecule). Thermal decomposition was observed at 240 °C and above.

[0094] The heat flow signal showed a distinct endothermic reaction around 160 °C, which is probably related to the melting of the anhydride of the said salt.

[0095] The DSC analysis results for the said Camp1 showed a broad endothermic reaction from 25 to 120 °C, which may probably be related to the loss of water. An endothermic reaction appeared around 164.0 °C, which is probably due to the melting of the potential anhydrous salt.

[0096] The chemical purity of the said Camp1 was measured by LCMS and was 100%, which confirmed the presence of API in the solid phase of Camp1.

[0097] The 1H-NMR analysis results for Camp1 are presented in Figure 19. The shift of the API resonance seen in the spectrum of Camp1 implies that proton transfer occurred with the acidic counterion in the free base form. The ratio of the said API:CI appeared as 1:1, and residual acetone was observed at 2.1 ppm.

[0098] Camp2 is a salt form obtained in an experiment where camphor-10-sulfonic acid was added so that the molar ratio of API:CI became 1:1 in an ACN solvent (Experiment ID: SSm60). Table 15 shows the experimental conditions for obtaining Camp2. Camp2 was physically stable when exposed to AAC (40 °C / 75% RH) for 2 days.

[0099]

Table 15

[0100] The TGA / TGMS analysis results for Camp2 showed a 1.6% mass loss from 25 to 100 °C due to water based on the MS signal (1.6% corresponds to 0.7 water molecules per salt molecule). Thermal decomposition was observed at temperatures above 240 °C.

[0101] The heat flow signal showed an endothermic reaction near 200 °C, which was probably related to the melting of the anhydride of the salt, followed by decomposition.

[0102] The DSC analysis results of Camp2 showed a broad endothermic reaction from 25 to 70 °C, which was probably related to the loss of water. An endothermic reaction was seen near about 208.9 °C, which was probably due to the melting of the anhydrous salt.

[0103] For the purpose of isolating the potential anhydride melting at 208.9 °C shown by the DSC results, a sample of Camp2 (experimental ID: SSm60) was dried under reduced pressure at 80 °C. The solid particles were then analyzed by HT-XRPD and TGMS. Camp2 remained in the solid particles and contained 1.9% water.

[0104] The chemical purity of Camp2 was 100% as measured by LCMS, which confirmed the presence of API in the solid phase of Camp2.

[0105] The 1H-NMR analysis results for Camp2 are shown in Figure 25. The shift of the API resonance seen in the spectrum of Camp2 implies that proton shift occurred with the acidic counterion in the free base form. The ratio of API:CI was 1:1.

[0106] Characterization analysis results of Mes2 The HT-XRPD analysis results can confirm the crystallization of Mes2 (experimental ID: SSm71) in the scale-up experiment. The high-resolution XRPD (HR-XRPD) pattern of Mes2 was not indexed.

[0107] The TGA / TGMS analysis results of dried Mes2 showed a gradual 4.0% mass loss between 25 and 110 °C, which is presumably due to water loss based on the MS signal (4.0% corresponds to 1.8 water molecules per salt molecule). The thermal decomposition of Mes2 was observed at temperatures above 240 °C.

[0108] The DSC analysis results of Mes2 showed one broad endothermic reaction related to water loss from 25 to 110 °C. The double endothermic reaction observed from 205 to 217 °C is presumably related to the melting process of the said salt. The chemical purity of Mes2 measured by LCMS was 99.6%, which confirmed the presence of API in the solid phase of Mes2.

[0109] The 1H-NMR analysis results of Mes2 are shown in Figure 30. The shift of the API resonance observed in the Mes2 spectrum implies that the proton transfer to the free base occurred with the acidic counterion. The API:CI ratio was 1:2.

[0110] Figure 31 shows the DVS measurement results for Mes2. The first moisture absorption in the half-cycle indicates that water absorption reached from a relative humidity of 95% to 29.6% (a 24.8% mass change appears with a change in relative humidity from 40 to 95%). Water was ejected during the desorption cycle from relative humidity 95 to 0%. The said water absorption could not maintain equilibrium with each step change in relative humidity (1-hour equilibrium standing at each step), which was the cause of the hysteresis that appeared between relative humidity 95 and 70% in the moisture absorption-desorption cycle. The water absorption was reversible.

[0111] The mass loss was approximately 12.2% at a relative humidity of 80%. As a result, the mesylate Mes2 showed moderate hygroscopicity in the European Pharmacopoeia hygroscopicity classification. There was no change in the solid form after the said DVS analysis.

[0112] Characterization analysis results of Camp2 According to the HT-XRPD analysis results, crystallization of Camp2 (experiment ID: SSm73) can be confirmed in the scale-up experiment. The high-resolution XRPD (HR-XRPD) pattern of Camp2 was not indexed.

[0113] The TGA / TGMS analysis results of dried Camp2 showed a 1.6% mass loss between 25 and 80 °C, which is presumably due to water loss based on the MS signal (1.6% corresponds to 0.7 water molecules per salt molecule). Thermal decomposition of Camp2 was observed at temperatures above 240 °C.

[0114] The DSC analysis results of Camp2 showed one broad endothermic reaction related to water loss between 25 and 80 °C. The second distinct endothermic reaction was observed at 208.6 °C, which is presumably related to the melting process of the anhydrous salt. The chemical purity of Camp2 measured by LCMS was 99.7%, which confirmed the presence of the API in the solid phase of Camp2.

[0115] The 1H-NMR analysis results of Camp2 are presented in Figure 36. The shift of the API resonance observed in the Camp2 spectrum implies that proton shift occurred with the free base with an acidic counterion. The API:CI ratio was 1:1.

[0116] Figure 37 shows the DVS measurement results for Camp2. The first moisture absorption in the half-cycle indicates that water absorption reached from a relative humidity of 95% to 15.6% (a 14.1% mass change appears with a change in relative humidity from 40% to 95%). The graph showing the mass change indicates that water absorption cannot reach equilibrium at relative humidities from 90% to 95%. Water was ejected during the desorption cycle from relative humidity 95% to 0%. In the last half of the absorption cycle (relative humidity 0% to 40% interval), the mass change was 6.6%. Water absorption was irreversible, which implies a change in morphology. A new form of Camp4 was confirmed in the solid phase by HT-XRPD (Figure 38).

[0117] The DVS analysis results show that the mass change is about 1.4% at a relative humidity of 80%, which implies that the substance is slightly hygroscopic according to the European Pharmacopoeia hygroscopicity classification.

[0118] Safety of the free base form Form A was exposed to 25°C / 60%RH and 40°C / 75%RH for 3 months of TGA analysis. The results are presented in Figures 40A and 40B. Mass losses of 4.1% and 4.4% were observed in the samples exposed to 25°C / 60%RH and 40°C / 75%RH for 3 months, respectively. The heat flow analysis results showed a broad endothermic reaction due to water loss at 140°C corresponding to the melting of Form A. Thermal decomposition was observed at 230°C in both samples.

[0119] Form A was exposed to 25°C / 60%RH and 40°C / 75%RH for 15 months of TGA analysis. The results are presented in Figures 41A and B. Mass losses of 3.3% and 3.7% were observed in the samples exposed to 25°C / 60%RH and 40°C / 75%RH for 15 months, respectively. The heat flow analysis results showed a broad endothermic reaction due to water loss at 140°C corresponding to the melting of Form A. Thermal decomposition was observed at 230°C in both samples.

[0120] Form A was exposed to 25°C / 60%RH and 40°C / 75%RH for 24 months of TGA analysis. The results are presented in Figures 42A and B. Mass losses of 5.3% and 5.2% were observed in the samples exposed to 25°C / 60%RH and 40°C / 75%RH for 24 months, respectively. The heat flow analysis results showed a broad endothermic reaction due to water loss at 140°C corresponding to the melting of Form A. No thermal decomposition occurred below 200°C.

[0121] After exposing the said Form A to 25°C / 60%RH (Experiment ID: GEN81) and 40°C / 75%RH (Experiment ID: GEN76) for 24 months, the chemical purities measured by LCMS were 87.2% and 90.7% respectively (Figures 43 and 44). This confirms that the compound deteriorated under both conditions.

[0122] Stability of mesylate According to the HT-XRPD analysis results, it can be confirmed that the conversion of the solid form occurred after Mes2 was exposed to 25 °C / 60% RH and 40 °C / 75% RH. The change occurred earlier at 40 °C / 75% RH than at 25 °C / 60% RH. There was no special difference in the powder pattern after the Mes2 sample was exposed to 25 °C / 60% RH for 3 months, but additional diffraction peaks were detected at 15.1 and 16.5° 2θ after 15 months. Probably the diffraction peaks seem to be caused by the change of other crystal forms, but the nature of the form was not known.

[0123] In the sample exposed to 40 °C / 75% RH, additional peaks were seen after 3 months. After 15 months, all the peaks of XRPD shifted. Although this was demonstrated with Mes4, the additional peaks still existed at 15.1 and 16.5° 2θ. After 24 months, the solids exposed to the two conditions were converted to a new form, which was named Mes6. Figure 45 shows the scale-up of Mes2 and the superimposition of the powder patterns after exposure to 25 °C / 60% RH and 40 °C / 75% RH for 1, 3, 15, and 24 months.

[0124] Figures 46A and 46B show the TGA analysis results after the Mes2 sample was exposed to 25 °C / 60% RH and 40 °C / 75% RH for 1 month. In the sample exposed to 25 °C / 60% RH for 1 month, 8.0% water loss occurred, and in the sample exposed to 40 °C / 75% RH for 1 month, 10.8% water loss occurred. Thermal decomposition occurred at 230 °C in both samples. Significant water absorption appeared compared to the initially measured moisture content of Mes2 in both samples (3.9%).

[0125] The water absorption also appeared after 3 months. Figures 47A and 47B show that the water losses after exposure to 25 °C / 60% RH and 40 °C / 75% RH for 3 months appeared at 7.7% and 11.2% respectively.

[0126] The thermal change of the sample after being exposed for 15 months under storage conditions showed very little mass loss and almost the same results as the above sample. Figures 48A and 48B show that the water loss after being exposed to 25°C / 60%RH and 40°C / 75%RH for 15 months was 6.5% and 9.6% respectively.

[0127] After 24 months, Mes2 showed mass losses of approximately 8.6% and 12.0% respectively after being incubated at 25°C / 60%RH and 40°C / 75%RH. Similar thermal changes were recorded in the heat flow signal. An endothermic reaction started at 200°C, which was understood to be due to the thermal decomposition of the mesylate.

[0128] After exposing Mes2 to 25°C / 60%RH (Experiment ID: GEN66) and 40°C / 75%RH (Experiment ID: GEN70) for 1 month, the chemical purities measured by LCMS were 100% and 99.7% respectively (Figures 50 and 51). This confirmed the presence of API in the solid phase of Mes2 with almost the same purity as the original Mes2 salt.

[0129] The chemical purities measured after exposing Mes2 to 25°C / 60%RH and 40°C / 75%RH for 3 months were 99.6% and 99.5% respectively (Figures 52 and 53).

[0130] The chemical purities of the samples after 15 months at 25°C / 60%RH and 40°C / 75%RH were 100% (Figures 54 and 55). The chemical purities of the samples after 24 months at 25°C / 60%RH and 40°C / 75%RH were 99.6% and 100% respectively (Figures 56 and 57).

[0131] Stability of camsylate The HT-XRPD analysis results confirmed that no solid form conversion occurred after exposing Camp2 to 25°C / 60%RH for 24 months.

[0132] However, at 40 °C / 75% RH, solid phase conversion was observed after 3 months. The new powder pattern was named Camp4 powder pattern. After 24 months, more conversion occurred, indicating the possibility of a mixture of camsylate and form E in the free base form. Figure 58 shows the superposition of the scale-up of Camp2 and the powder patterns after exposure to 25 °C / 60% RH and 40 °C / 75% RH for 1, 3, 15, and 24 months.

[0133] There was no particular difference in the powder pattern after exposing the Mes2 sample to 25 °C / 60% RH for 3 months, but additional diffraction peaks were detected at 15.1 and 16.5° 2θ after 15 months. Presumably, the diffraction peaks are likely caused by changes in other crystal forms, but the nature of the form is unknown.

[0134] Additional peaks were seen after 3 months in the samples exposed to 40 °C / 75% RH. After 15 months, all the peaks of XRPD shifted, which was designated as Mes4, but the additional peaks still existed at 15.1 and 16.5° 2θ. After 24 months, the solids exposed to the two conditions were converted to a new form, which was named Mes6. Figure 45 shows the superposition of the scale-up of Mes2 and the powder patterns after exposure to 25 °C / 60% RH and 40 °C / 75% RH for 1, 3, 15, and 24 months.

[0135] Figures 59A and 59B show the TGA analysis results of Camp2 after exposure to 25 °C / 60% RH and 40 °C / 75% RH for 1 month. A 1.6% mass loss appeared in the sample exposed to 25 °C / 60% RH for 1 month, and a 1.8% mass loss was shown in the sample exposed to 40 °C / 75% RH for 1 month. Thermal decomposition occurred at 240 °C or higher in both samples.

[0136] The Camp2 samples incubated at 25°C / 60%RH for 3 months showed the same water content as that recorded after 1 month, which means that there was no significant water absorption under the said conditions. However, a solid-state conversion from Camp2 to Camp4 was observed for Camp2 at 40°C / 75%RH. As a result, the TGA analysis results showed a water content of 7.1% (corresponding to 3.3 water molecules per camcillate molecule). The thermal changes recorded by the heat flow signal showed a broad endothermic reaction due to the elution of water, followed by an endothermic reaction at 150°C caused by the melting of the anhydride (200°C for Camp2).

[0137] The Camp2 samples incubated at 25°C / 60%RH for 24 months showed a 1.9% mass loss, which is the same as the mass loss measured by the said formula (Figure 62A). This result means that no significant water absorption occurred under the said conditions. The TGA analysis results after exposure to 40°C / 75%RH for 24 months showed a moisture content of 6.7% (corresponding to 3.2 water molecules per camcillate molecule). The thermal changes recorded by the heat flow signal showed a broad endothermic reaction due to the elution of water, followed by an endothermic reaction at 150°C caused by the melting of the anhydride (200°C for Camp2).

[0138] After exposing the said Camp2 to 25°C / 60%RH (Experiment ID: GEN50) and 40°C / 75%RH (Experiment ID: GEN54) for 1 month, the chemical purity measured by LCMS was 99.8% for both samples (Figure 63, Figure 64). This means that the API is present in the solid phase of Camp2. The chemical purity after 3 months can be compared with that of the 1-month samples (Figure 65, Figure 66).

[0139] The chemical purity of Camp2 after exposure to 25°C / 60%RH for 15 months was 99.5% (Figure 67), which means that Camp2 is stable for at least 15 months under the said conditions. The chemical purity of the Camp4 samples after 15 months at 40°C / 75%RH was 93.7% (Figure 68). This means that Camp2 is converted to Camp4 at 40°C / 75%RH.

[0140] After exposing Camp2 to 25°C / 60%RH for 24 months, the chemical purity was 100% (Figure 69), which confirms that Camp2 is stable for at least 24 months under the said conditions. After exposing the camsylate to 40°C / 75%RH for 24 months, the chemical purity was 88.6%, which means that when Camp2 is converted into a different form, chemical deterioration occurs at 40°C / 75%RH.

[0141] Although the present invention has been described by citing the special embodiments and the like, it is obvious that various modifications can be made without departing from the technical idea of the present invention. Therefore, it is a well-known fact that the present invention is not limited to the described embodiments, but also includes those based on the claims and equivalent substances.

Claims

1. A mesylate salt made of methanesulfonic acid and a triazolopyrazine derivative represented by the following Chemical Formula 1. 【Chemical 1】

2. The mesylate salt according to Claim 1, wherein the mesylate salt is an S optical isomer of the triazolopyrazine derivative of Chemical Formula 1.

3. The mesylate salt according to Claim 2, wherein the mesylate salt is characterized in that the triazolopyrazine derivative of Chemical Formula 1 is optically pure.

4. The mesylate salt according to any one of Claims 1 to 3, wherein the mesylate salt is any one of Mes1, Mes2, and Mes3.

5. The mesylate salt according to Claim 4, wherein the mesylate salt is Mes2.

6. The mesylate salt according to Claim 1, wherein the mesylate salt is physically stable at 20 - 50 °C and a relative humidity of 35 - 80% for at least 2 days.

7. The mesylate salt according to Claim 6, wherein the API purity is at least 95%.

8. The mesylate salt according to Claim 7, wherein the purity of the API is 100%.

9. The mesylate salt according to Claim 1, wherein the high-efficiency XRPD (HT-XRPD) pattern shows characteristic peaks at 15.5 to 16.0 °2θ, 17.5 to 18.0 °2θ, and 21.5 to 22.0 °2θ.

10. The mesylate salt according to Claim 1, wherein the HT-XRPD pattern of the mesylate salt has substantially the following characteristic peaks. Or,

11. It relates to a method for producing the mesylate salt of Claim 1, and the production method includes: a) A step of adding a compound of the following Chemical Formula 1 to a reactor containing a solvent; 【Chemical 1】 b) A step of stirring the compound and the solvent in the reactor; c) A step of adding methanesulfonic acid to the solution in step b); d) A step of cooling the solution in step c) to obtain a precipitate, and the method for producing a mesylate salt is characterized by including these steps.

12. The production method according to Claim 11, wherein the solvent contains any one selected from acetonitrile, acetone, 1,2-dimethoxyethane, n-heptane, isopropyl alcohol, water, or tetrahydrofuran (THF).

13. The manufacturing method according to claim 12, wherein the methanesulfonic acid added in the step b) is added in an equivalent amount of 1:1.5 to 1:2.5 with respect to the compound of the chemical formula 1.

14. The manufacturing method according to claim 13, wherein the equivalent amount is 1:1.9 to 1:2.3, and the solvent is acetonitrile.

15. The manufacturing method according to any one of claims 11 to 14, wherein the step b) is carried out at 45 to 55 ° C for at least 1 hour.

16. A pharmaceutical composition comprising the mesylate of claim 1 and a pharmaceutically acceptable carrier.

17. The pharmaceutical composition according to claim 16, wherein the pharmaceutically acceptable carrier is one or more selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, green powder, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, amorphous cellulose, polyvinylpyrrolidone, cellulose, water, shellac, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

18. The pharmaceutical composition according to claim 17, wherein the pharmaceutical composition further comprises one or more selected from the group consisting of a lubricant, a wetting agent, a sweetening agent, a spice, an emulsifier, a suspending agent and a preservative.

19. As a method for suppressing the activity of c-Met kinase, the method is characterized by injecting an effective amount of the camsylate of claim 1 into a subject.

20. The method according to claim 19, wherein the subject exhibits a proliferative disorder.

21. The method according to claim 20, wherein the proliferative disorder is any one selected from lung cancer, colon cancer, pancreatic cancer, rectal cancer, ovarian cancer, renal cell carcinoma, prostate cancer and brain tumor.

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