Salt forms of organic compounds

Abstract

Salt compounds of 4-(2-((4-bromophenyl)thio)acetamido)-1-phenethyl-1H-pyrazole-3-carboxylic acid are disclosed. Also disclosed are methods for making the salt compounds and methods for formulating the salt compounds into dosage forms for clinical use. Also disclosed are methods for formulating the salt compounds and salt compound formulations into dosage forms for clinical use.

Description

Technical Field

[0001] The present disclosure relates to various formulations and compositions comprising salt compounds useful as inhibitors of FAF-1. Also disclosed is a method for preparing the salt compounds.

Background Art

[0002] Ischemia means a decrease in blood flow to an organ, tissue or region thereof caused by constriction or occlusion of one or more blood vessels. Once ischemia occurs, various sequelae that develop due to damage to nerve cells continue, even if reperfusion is rapid. Such ischemia frequently occurs in coronary artery disease, cardiovascular disease, angina, headache, or other symptoms associated with vascular occlusion or constriction, ultimately resulting in irreversible damage, i.e., necrosis of cells or tissues.

[0003] Ischemic diseases such as myocardial infarction, arrhythmia or heart failure caused by cell damage and dysfunction during ischemia-reperfusion have a high morbidity, high mortality and low complete cure rate, so in this field, basic research and clinical studies have been ongoing for 50 years [Wang, Q. D. et al., Cardiovasc. Res. 55:25-37, 2002]. In particular, ischemia-reperfusion injury is accompanied by various physiological mechanisms including changes in metabolism, immune response and ion homeostasis, generation of oxygen free radicals, etc., so studies are being advanced in various fields related to immunomodulation, suppression of cell death, regulation of ion channels, etc. [Hearse, D. J. et al., Mol. Cell. Biochem. 186:177-184, 1998]. Based on such mechanism research, many therapeutics and surgical procedures focusing on new action sites have been developed so far, but the technology for protecting cardiomyocytes from ischemia-reperfusion injury has not yet been commercialized. Therefore, there is a need for a drug for preventing and treating ischemic heart disease or a drug for preventing heart disease that can delay the progression of ischemic damage to cardiomyocytes and reduce reperfusion-induced injury.

[0004] Furthermore, it has been shown that when ischemia is relieved by the restoration of blood flow, the production of reactive oxygen species (ROS) is promoted, leading to a significant decrease in glutathione and causing more serious disease. Similar diseases are observed when blood flow is stopped or restored during various techniques of organ transplantation surgery for various types of organs such as the heart, liver, lungs, pancreas, or blood vessels, and are also problematic in organ dissection and removal. Reactive oxygen species and active free radicals suspected to cause disease are detected in the cytoplasm and organelles of tissue cells, particularly in mitochondria, which produce ATP as the cell's primary energy source. In mitochondria, the above-mentioned active molecules are released mainly via the respiratory chain, and their concentrations are observed to increase significantly during ischemic perfusion.

[0005] In this regard, ischemia leads to cell death or necrosis, and cell death occurring particularly after reperfusion is a major cause of tissue damage. Therefore, ischemic cell death is the cause of various ischemic diseases, such as cerebral ischemia, cardiac ischemia, diabetic cardiovascular disease, heart failure, myocardial hypertrophy, retinal ischemia, ischemic colitis, and ischemic acute renal failure.

[0006] In cerebral ischemia, depletion of energy sources due to reduced blood supply induces ischemic cell death. This ischemic cell death then excessively activates cell membrane receptors, followed by various biochemical changes on the extracellular and intracellular sides, including the accumulation of glutamate and calcium, and damage to lipids, proteins, and nucleic acids, ultimately leading to brain tissue damage (Liu, PK, J. Biomed. Sci. 10:4-13, 2003; Upton, R, Physiol. Rev. 79:1431-1568, 1999; and Renolleau, S. et al., Stroke 29:1454-1460, 1998).

[0007] In ischemic heart diseases such as myocardial infarction, heart failure, and arrhythmias, ischemic cell death has been reported to occur through the activation of lipid enzymes that cause damage to the cell membrane, followed by changes in pH and calcium transport [Ferrari, R. Rev. Port. Cardiol. 5:7-20, 2000; Webster, KA et al., J. Clin. Invest. 104:239-252, 1999; Katz, AM et al., J. Mol Cell. Cardiol.2:11-20, 1985; and Vandeplassche, G. et al., Basic Res. Cardiol. 85:384-391, 1990]. In retinal ischemia, glutamate-mediated retinal cell death is known to be mediated by ischemic cell death [Napper, GA et al., Vis. Neurosci. 16:149-158, 1999]. Insufficient blood supply to the colon causes ischemic cell death, and subsequently, occlusive arterial damage due to cell necrosis and hemodynamic impairment leads to ischemic colitis as an ischemic disease [Saegesser, F. et al., Pathobiol. Annu. 9:303-337, 1979].

[0008] On the other hand, minocycline, a tetracycline antibiotic that inhibits ischemic cell death, is known to be effective against ischemic diseases such as cerebral infarction [Yrjanheikki, J. et al., Proc. Natl. Acad. Sci. USA 96: 13496-1 3500, 1999], myocardial infarction [Scarabelli, TM et al., J.Am. Coll. Cardiol. 43:865-874, 2004], and ischemic acute renal failure [Wang, J. et al., J. Biol. Chem. 279:19948-19954, 2004], suggesting that ischemic cell death is the cause of these diseases.

[0009] Furthermore, ischemia-induced neuronal damage or cell death is known to be a major cause of various neurological diseases such as Alzheimer's disease, Parkinson's disease, glaucoma, and diabetic neuropathy, as well as pathologies resulting from stroke, head injury, and neonatal hypoxia [GJ Zoppo et al., Drugs 54, 9 (1997); I. Sziraki et al., Neurosci. 85, 1101 (1998)]. [Overview of the project] [Means for solving the problem]

[0010] A salt compound having formula (2) (see below) is disclosed.

[0011] [ka]

[0012] Formula (2): [n is 1, 2, or 3, m can be a non-integer from 0 to 3, and is typically 0, 0.5, 1, 2, or 3. "Sol" is a solvent molecule, which may be, for example, water or a C2-C4 alcohol. X + [ is a cation, which may be, for example, a potassium ion, a sodium ion, a calcium ion, a magnesium ion, an ammonium ion, or a substituted ammonium ion.] A salt compound having [

[0013] The salt compounds of formula 2 can be prepared by treating the free base or zwitterion (described later) of compound 1 with, for example, potassium hydroxide, sodium hydroxide, L-arginine, calcium hydroxide, N,N,N-trimethylglycine, ammonium hydroxide, magnesium hydroxide, choline, diethylamine, L-lysine, N,N'-dibenzylethylenediamine, N-ethylglucamine, calcium acetate, 1-(2-hydroxyethyl)pyrrolidine, N-(phenylmethyl)benzeneethanamine, ammonia, magnesium acetate, N-methylglucamine, tromethamine, 4-(2-hydroxyethyl)morpholine, 2-(diethylamino)ethanol, or 2-dimethylaminoethanol. [Brief explanation of the drawing]

[0014] [Figure 1] Figure 1 shows the XRPD diffraction of samples prepared with potassium hydroxide in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0015] [Figure 2] Figure 2 shows the XRPD diffraction of samples prepared with sodium hydroxide in various solvents. 1 is from ethanol, 2 is from diisopropyl ether, 3 is from 4-methylpentan-2-one, and 4 is from free acid.

[0016] [Figure 3] Figure 3 shows the XRPD diffraction of samples prepared with L-arginine in various solvents. 2 is from diisopropyl ether, 3 is from 4-methylpentan-2-one, and 4 is from free acid.

[0017] [Figure 4] Figure 4 shows the XRPD diffraction of samples prepared with calcium hydroxide in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0018] [Figure 5] Figure 5 shows the XRPD diffractograms of samples prepared with N,N,N-trimethylglycine in various solvents. 1 is from 4-methyl-pentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from the free acid.

[0019] [Figure 6] Figure 6 shows the XRPD diffractograms of samples prepared with ammonium hydroxide in various solvents. 1 is from 4-methyl-pentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from the free acid.

[0020] [Figure 7] Figure 7 shows the XRPD diffractograms of samples prepared with magnesium hydroxide in various solvents. 1 is from 4-methyl-pentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from the free acid.

[0021] [Figure 8] Figure 8 shows the XRPD diffractograms of samples prepared with choline in various solvents. 1 is from 4-methyl-pentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from the free acid.

[0022] [Figure 9] Figure 9 shows the XRPD diffractograms of samples prepared with diethylamine in various solvents. 1 is from 4-methyl-pentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from the free acid.

[0023] [Figure 10] Figure 10 shows the XRPD diffractograms of samples prepared with L-lysine in various solvents. 1 is from 4-methyl-pentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from the free acid.

[0024] [Figure 11] Figure 11 shows the XRPD diffraction of samples prepared with N,N'-dibenzylethylenediamine in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0025] [Figure 12] Figure 12 shows XRPD diffractions prepared with N-ethylglucamine in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0026] [Figure 13] Figure 13 shows the XRPD diffraction of calcium acetate-prepared samples in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0027] [Figure 14] Figure 14 shows the XRPD diffraction of samples prepared with N-(phenylmethyl)benzeneethanamine in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0028] [Figure 15] Figure 15 shows the XRPD diffraction of ammonia-prepared samples in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0029] [Figure 16] Figure 16 shows the XRPD diffraction of magnesium acetate-prepared samples in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0030] [Figure 17]Figure 17 shows the XRPD diffraction of samples prepared with N-methylglucamine in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0031] [Figure 18] Figure 18 shows the XRPD diffraction of samples prepared with tromethamine in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0032] [Figure 19] Figure 19 shows the XRPD diffraction of samples prepared with 4-(2-hydroxyethyl)morpholine in various solvents. 1 is from 4-methylpentan-2-one, 2 is from diisopropyl ether, 3 is from ethanol, and 4 is from free acid.

[0033] [Figure 20] Figure 20 shows the PLM analysis of potassium salt formula 2 from the second preparation of KM-819.

[0034] [Figure 21] Figure 21 shows the TGA analysis of potassium salt formula 2 from the second preparation of KM-819.

[0035] [Figure 22] Figure 22 shows differential scanning calorimetry (DSC) analysis of potassium salt formula 2 from the second preparation of KM-819.

[0036] [Figure 23A-E] Figures 23A-23E show the HSM analysis of potassium salt formula 2 from the second preparation of KM-819. 23A: Initial conditions for the potassium salt, 23B: Birefringence loss at 127°C, 23C: Initial melting at 154°C, 23D: Secondary melting at 212°C, 23E: Recrystallization.

[0037] [Figure 24]Figure 24 shows the Fourier transform infrared (FT-IR) analysis of potassium salt formula 2 from the second preparation of KM-819.

[0038] [Figure 25] Figure 25 shows the potassium salt particle size distribution (PSD) of formula 2 from the second preparation of KM-819.

[0039] [Figure 26] Figure 26 shows the DVS change in the mass plot of potassium salt formula 2 from the second preparation of KM-819.

[0040] [Figure 27] Figure 27 shows the DVS isothermal plot of potassium salt formula 2 from the second preparation of KM-819.

[0041] [Figure 28] Figure 28 shows the PLM analysis of the sodium salt from the second preparation of KM-819.

[0042] [Figure 29A-C] Figures 29A-29C show the HSM analysis of the sodium salt from the second preparation of KM-819. 29A: Prepared salt, 29B: Melted at 136°C, 29C: Recrystallized.

[0043] [Figure 30] Figure 30 shows the TGA analysis of the sodium salt from the second preparation of KM-819.

[0044] [Figure 31] Figure 31 shows the DSC analysis of the sodium salt from the second preparation of KM-819.

[0045] [Figure 32] Figure 32 shows the FT-IR analysis of the sodium salt from the second preparation of KM-819.

[0046] [Figure 33]Figure 33 shows the Sympatec particle size distribution analysis of the sodium salt from the second preparation of KM-819.

[0047] [Figure 34] Figure 34 shows the DVS change in the mass plot of the sodium salt from the second preparation of KM-819.

[0048] [Figure 35] Figure 35 shows the DVS isothermal plot of the sodium salt from the second preparation of KM-819.

[0049] [Figure 36] Figure 36 shows the PLM analysis of the diethylamine salt formula 2 from the second preparation of KM-819.

[0050] [Figure 37] Figure 37 shows the TGA analysis of the diethylamine salt formula 2 from the second preparation of KM-819.

[0051] [Figure 38] Figure 38 shows the DSC analysis of the diethylamine salt formula 2 from the second preparation of KM-819.

[0052] [Figure 39A-E] Figures 39A-E show the HSM analysis of the diethylamine salt formula 2 from the second preparation of KM-819. 39A: Prepared diethylamine salt, 39B: Initial melting at 154°C, 39C: Particle motion at 200°C, 39D: Secondary melting at 209°C, 39E: Recrystallization.

[0053] [Figure 40] Figure 40 shows the FT-IR analysis of the diethylamine salt formula 2 from the second preparation of KM-819.

[0054] [Figure 41] Figure 41 shows the particle size distribution of the diethylamine salt formula 2 of KM-819 from the second preparation.

[0055] [Figure 42] Figure 42 shows the DVS change in the mass plot of formula 2 of the diethylamine salt from the second preparation of KM-819.

[0056] [Figure 43] Figure 43 shows the DVS isothermal plot of the diethylamine salt formula 2 from the second preparation of KM-819.

[0057] [Figure 44] Figure 44 shows the alignment of five XRPD diffraction patterns of salts obtained during screening. The spectra are aligned to a 2-theta scale.

[0058] [Figure 45] Figure 45 shows alignments of five different 1H NMR analyses observed in samples from salt screening. The spectra are aligned via the standard peak of DMSO. The pattern numbering corresponds to the XRPD pattern numbering, with salts showing XRPD pattern 1 often showing NMR pattern 1. [Modes for carrying out the invention]

[0059] Aminopyrazole derivatives have been disclosed to inhibit ischemic cell death and can therefore be used as agents for the prevention and treatment of ischemic diseases mediated by ischemic cell death, such as cerebral ischemia, cardiac ischemia, diabetic cardiovascular disease, heart failure, myocardial hypertrophy, retinal ischemia, ischemic colitis, ischemic acute renal failure, stroke, traumatic brain injury, Alzheimer's disease, Parkinson's disease, neonatal hypoxia, glaucoma, and diabetic neuropathy, as well as as agents for protecting organs during transplant surgery.

[0060] Compound 1 (KM-819) is a novel aminopyrazole derivative useful for the treatment of Parkinson's disease. Disclosed Compound 1 (KM-819) can be synthesized as described in W02008 / 051047 (which, for all purposes, is incorporated herein by reference in its entirety) and provides a white crystalline powder. The initial analysis of Compound 1 (KM-819) was performed as a free acid or zwitterion to provide reference data that would allow for a better understanding of the material and enable comparisons between Compound 1 (KM-819) and any prepared salts.

[0061] Those skilled in the art will understand that compound 1 contains both a carboxylate group capable of forming an anion and a nitrogen center capable of forming a cationic quaternary amine. Thus, “compound 1” can refer to either the free acid or zwitterionic form of the compound, depending on the pH of the solution of compound 1.

[0062] [ka]

[0063] The solubility of compound 1 (KM-00819) was evaluated using water and a diverse range of organic solvents. A list of solvents suitable for use during salt screening was determined from this evaluation. Upon completion of solvent screening, 1.1 equivalents of sodium hydroxide (aqueous solution) were added to each sample to test its ability to form a salt. Upon addition of hydroxide, turbid precipitates formed from some samples, indicating that salt formation had likely occurred, and XRPD analysis of the solids isolated from the salt formations revealed five distinct diffraction patterns. (See, for example, Example 8 and Figures 10-19 and 44.) Using 1H NMR analysis of the samples exhibiting diffraction patterns, the most suitable solvents for preparing the salt forms of KM-819 were determined, as described below. Figure 45 shows the alignment of representative 1H NMR analyses of samples from the salt screening.

[0064] <Salt screening> Salt screening was performed for each experiment using three solvents, 22 bases, and approximately 25 mg of compound 1 (KM-819), added in a 1:1.1 (free acid:base) ratio. After preparation, the samples were allowed to stand for 5 days before filtration and analysis by XRPD. If the salts remained completely soluble in the solvent, the solvent was slowly evaporated from the sample.

[0065] The water solubility of salts exhibiting new XRPD patterns was evaluated, and those that showed complete or partial dissolution after overnight shaking at 50°C (concentration of 1.25 mg / ml) were further analyzed by 1H NMR. Some salts showed pleomorphism as indicated by differences in XRPD patterns and 1H NMR chemical shifts.

[0066] Disclosed herein are novel pharmaceutically acceptable solid forms of KM-819, their salts, and methods for preparing them. These forms can be used by preparing salts or bases and preparing their formulations for clinical use.

[0067] Disclosed herein are novel pharmaceutically acceptable salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid (KM-819) in crystalline or amorphous forms and methods for their preparation. These salts can be used to prepare other salts or free base forms of KM-819 and to prepare formulations thereof for clinical use.

[0068] The present invention relates to salt and / or solvate (hydrate) compounds according to the following formula 2.

[0069] [ka]

[0070] In equation 2, n is an integer of 1, 2, or 3. m can be a non-integer from 0 to 3, such as 0.5 or 1.5. "Sol" is a solvent molecule, which may be, for example, water or a C2-C4 alcohol. X + is a cation, and may be, for example, a potassium ion, a sodium ion, a calcium ion, a magnesium ion, an ammonium ion, or a substituted ammonium ion.

[0071] The salt compounds of formula 2 can be prepared by treating the free base or zwitterion of compound 1 with, for example, potassium hydroxide, sodium hydroxide, L-arginine, calcium hydroxide, N,N,N-trimethylglycine, ammonium hydroxide, magnesium hydroxide, choline, diethylamine, L-lysine, N,N'-dibenzylethylenediamine, N-ethylglucamine, calcium acetate, 1-(2-hydroxyethyl)pyrrolidine, N-(phenylmethyl)benzeneethanamine, ammonia, magnesium acetate, N-methylglucamine, tromethamine, 4-(2-hydroxyethyl)morpholine, 2-(diethylamino)ethanol, or 2-dimethylaminoethanol.

[0072] In some examples of the salt compounds of formula 2, m may be 0.5 or 1. In such examples, X + This may be a potassium ion, a sodium ion, or a quaternary methylamine or quaternary ethylamine.

[0073] Salt compound 2 may also be in the form of solvates such as those containing, alternatively or additionally, water, ethanol, or diisopropyl ether, or a mixture of any two or three of these. The solvent molecules may be present in non-integer ratios with respect to either or both water molecules and compound 1 ions, for example, 0.1, 0.2, or 0.5 solvent molecules per compound 1 ion. The solvent molecules may be present in integer ratios with respect to either or both water molecules and compound 1 ions, for example, 1 or 2 solvent molecules per compound 1 ion.

[0074] Also, equation (2):

[0075] [ka]

[0076] [In the formula, n is 1, 2, or 3. m ranges from 0 to 3. X + It is a positive ion, A method for preparing a salt compound having "Sol" as a solvent molecule, i) Dissolve the free acid form of the compound of formula 2 in an organic solvent or an organic solvent mixed with water; ii) Adding an excess amount of base in a quantity exceeding the stoichiometric amount required to titrate the free acid compound of formula 2 to form a precipitate of the salt compound of formula 2; and iii) Collect the precipitate to obtain salt compound 2. Methods including the following are also disclosed.

[0077] In this method, the organic solvent may be 1,1-dimethoxyethane, acetonitrile, ethanol, 1,2-dichloroethane, benzonitrile, ethyl acetate, 1,4-dioxane, anisole, heptane, 2-butanol, cumene, hexane, 2-propanol, cyclohexane, isopropyl acetate, 4-methylpentan-2-one, dichloromethane, methanol, acetone, diisopropyl ether (DIPE), isobutyl acetate, tetralin, toluene, methyl ethyl ketone (MEK), N-methylpyrrolidone, tert-butyl methyl ether (TMBE), nitromethane, pyridine, or tetrahydrofuran, or a mixture of any two or three of these.

[0078] In some embodiments of the method, the organic solvent may be ethanol or diisopropyl ether (DIPE).

[0079] In some embodiments of the method, the base may be sodium hydroxide, potassium hydroxide, magnesium hydroxide, magnesium acetate, ammonia, a salt of a quaternary dimethylamine, or a salt of a quaternary diethylamine.

[0080] A combination of ethanol or DIPE as the organic solvent and sodium hydroxide, potassium hydroxide, or a salt of quaternary dimethylamine or quaternary diethylamine as the base may be used.

[0081] In some cases, ethanol or DIPE is used as the organic solvent.

[0082] Water may be mixed with organic solvents, and in some cases, water may be mixed with ethanol or DIPE.

[0083] In any embodiment in which water is mixed with an organic solvent, for example, water may be mixed with a polar organic solvent, and the ratio of water to the polar organic solvent may range from 5:1 to 10:0.1.

[0084] Free base (zwitterionic) compound 1 may be dissolved in unbuffered water, a range of organic solvents, a mixture of organic solvents, and a mixture of solvent and unbuffered water. The solvents evaluated were 1,1-dimethoxyethane, acetonitrile, ethanol, 1,2-dichloroethane, benzonitrile, ethyl acetate, 1,4-dioxane, anisole, heptane, 2-butanol, cumene, hexane, 2-propanol, cyclohexane, isopropyl acetate, 4-methylpentan-2-one, dichloromethane, methanol, acetone, diisopropyl ether (DIPE), isobutyl acetate, tetralin, toluene, methyl ethyl ketone, N-methylpyrrolidone, tert-butyl methyl ether, nitromethane, pyridine, and tetrahydrofuran. The results of the solubility tests are shown in Table 2 below.

[0085] The organic solvent may be used as a pure solvent or a mixture of two, three, or more organic solvents. Water alone may be used as the solvent for the free base (zwitterionic) compound 1, or water may be mixed with one or more organic solvents. Preferably, a pure polar organic solvent or a solvent mixture, or a mixture of polar organic solvents and water, is used to dissolve the free base or zwitterionic compound 1.

[0086] In the case of a two-component mixture, the ratio of water to the organic solvent (preferably a polar organic solvent) in the solvent mixture may be 1:10 to 1:0.1, or 1:5 to 1:0.1, or 1:2-1:0.1, or 1:2-1:0.5, or about 1:1.

[0087] <Salt formation> A general method for preparing the salt compounds of this disclosure is illustrated in the following scheme.

[0088] Scheme 1 demonstrates the synthesis of the salt form following a general route utilizing well-established chemistry.

[0089] [ka]

[0090] Free acid compound 1 was weighed and added to a container, followed by the addition of a solvent. Then, approximately 1.1 equivalents of base, prepared as a 1 M stock solution in the solvent, were added. Turbidity of the sample upon base addition indicates that salt formation is occurring. After standing (at room temperature for several hours), the sample was filtered, dried under vacuum, and then characterized in various ways.

[0091] The bases used to form the salt may be potassium hydroxide, sodium hydroxide, L-arginine, calcium hydroxide, N,N,N-trimethylglycine, ammonium hydroxide, magnesium hydroxide, choline, diethylamine, L-lysine, N,N'-dibenzylethylenediamine, N-ethylglucamine, calcium acetate, 1-(2-hydroxyethyl)pyrrolidine, N-(phenylmethyl)benzeneethanamine, ammonia, magnesium acetate, N-methylglucamine, tromethamine, 4-(2-hydroxyethyl)morpholine, 2-(diethylamino)ethanol, or 2-dimethylaminoethanol.

[0092] Many organic compounds exist in different solid forms, which may be amorphous or crystalline.

[0093] The ability of a compound to crystallize in different crystalline phases is called polymorphism. The term polymorph may also include amorphous phases (disorder), hydrates (water present in the crystal lattice), and solvates (solvents other than water present in the crystal lattice).

[0094] Because different crystal transformations result in different crystal structures and different free energies, polymorphs exhibit different physicochemical properties such as melting point, density, solubility, chemical stability, and ultimately, bioavailability.

[0095] Examples of preferred salts of salt compound 2 of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid are as follows:

[0096] 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and potassium hydroxide salts,

[0097] 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and salts of sodium hydroxide,

[0098] 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and salts of ammonium hydroxide,

[0099] 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and salts of choline,

[0100] 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and diethylamine salts,

[0101] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and L-lysine,

[0102] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and N,N'-dibenzylethylenediamine,

[0103] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and N-ethylglucamine,

[0104] 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and salts of calcium acetate,

[0105] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and N-(phenylmethyl)benzeneethaneamine,

[0106] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and magnesium acetate,

[0107] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and N-methylglucamine,

[0108] 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and tromethamine salts,

[0109] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and 4-(2-hydroxyethyl)morpholine.

[0110] Many solids isolated from the salt formation stage exhibited diffraction patterns in XRPD analysis (indicating crystalline solids), some showing different diffraction patterns from those of free acid compound 1. In many cases, 1H NMR analysis of the crystalline solids indicated that the -COOH group in compound 1 was ionized.

[0111] A suitable solvent for salt formation is one that completely dissolves free acid compound 1, and a suitable base for preparing salt compound 2 from compound 1 is one that completely replaces the hydrogen atoms of the carboxylate in compound 1.

[0112] [Table 1] [Examples]

[0113] The following embodiments describe the preparation and detailed characterization of representative embodiments.

[0114] 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid (compound 1) was synthesized as described in WO2008 / 051047 to obtain a white crystalline powder.

[0115] The samples for the examples were synthesized by the disclosed methods (Examples 1, 2, 4, and 5) and analyzed by XRPD, 1H NMR, HPLC for chemical purity and solubility.

[0116] <Equipment used> Perkin Elmer PYRIS 1 DSC using a 40 μL aluminum dish (with vent). Data acquisition and analysis were performed using Perkin Elmer control and analysis software, version 11.0.2.0468.

[0117] A Bruker 400 Avance spectrometer equipped with a 5mm QNP probe was used. Analysis was performed using an ACD Laboratories 1D NMR processor, version 12.01, and instrument control and data acquisition were performed using Top Spin version 1.3.

[0118] Jasco 420 FTIR with attenuated total internal reflection (ATR) module. Analysis and data acquisition were performed using Jasco Spectra Manager software, version 1.51.00 (build 1).

[0119] For polarized optical microscopy, an Olympus BX 53 microscope with six objective lenses (2.5x, 4x, 10x, 20x, 40x, and 100x) and a 1 / 10λ waveplate. A Sony ICX252 progressive scan interline 3.3MP CCD camera. The microscope also features a Linkam LTS420 heating / freezing stage.

[0120] PLM: Data analysis and image acquisition using Qcapture-Pro version 7 imaging software.

[0121] HSM: Linksys 32DV temperature control and digital video capture software for data analysis and image acquisition.

[0122] A Bruker-AXS D8 Advance XRPD was used with a 9mm cavity and a flat sample holder. Instrument control and data acquisition were performed using a PC with Diffrac Plus XRD Commander control software version 2.6.1, and the recorded data was analyzed using Eva version 18.0.0.0.

[0123] This is a dynamic water vapor adsorption system specific to SMS DVS, using DVS-specific control software version 1.0.6.0. Data analysis was performed using the DVS analysis suite version 7.0.13.1 macro program, which is built into Microsoft Excel. The analysis was performed as wt% change from 0 to 90% RH, and isothermal plots were also examined.

[0124] A Perkin Elmer TGA PYRIS 1 was used with an aluminum dish (with vents) in a ceramic crucible. Data analysis and acquisition were performed using Perkin Elmer control and analysis software, version 11.0.2.0468.

[0125] Thermo-Fisher iCAP 6500 ICP-OES using iTEVA software.

[0126] The Metrohm 852 Titranto combines KF units for volumetric and coulometric measurements. All samples were analyzed using the volumetric Karl Fischer module.

[0127] A Waters-Alliance 2695 HPLC spectrometer equipped with a PDA2996 probe. System control and processing were performed using Empower 3 software build 3471.

[0128] Heidolph Titramax 1000 with heating module.

[0129] Example 1: Potassium salt of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid 1500 μL solventBefore adding, add approximately 25 mg of compound 1 (KM-00819 )of The sample was weighed into a 2 mL HPLC vial. To the resulting slurry, 1.1 equivalents of potassium hydroxide (at a 1 M concentration) in 60 μl of water were added. The sample was left to stand for 5 days using an 8-hour cycle (4 hours at room temperature, followed by 4 hours at 50°C). After standing, the sample was re-examined, then filtered and dried under vacuum.

[0130] Example 2: 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylate calcium salt.

[0131] As shown in Table 1, approximately 25 mg of compound 1 (KM-00819) was weighed into a 2 mL HPLC vial before adding 1500 μL of solvent. To the resulting slurry, 1.1 equivalents of calcium hydroxide (5.6) were added. μg The solid was added. The sample was left to stand for 5 days using an 8-hour cycle (4 hours at room temperature followed by 4 hours at 50°C). After standing, the sample was re-examined, then filtered and dried under vacuum.

[0132] Example 3: Additional salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid.

[0133] Salts of the following compounds were prepared by the same method as in Example 1 or Example 2.

[0134] [Table 2] JPEG0007834307000008.jpg205152 JPEG0007834307000009.jpg212152 JPEG0007834307000010.jpg130153

[0135] Example 4: 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylate sodium salt.

[0136] Before adding 60 mL of DIPE to each container, approximately 1 g of compound 1 was weighed into a 3 × 100 mL vial. To the resulting suspension, 1.1 equivalents of sodium hydroxide, prepared as a 1 M stock solution in water, were added. Upon addition of the base, the reaction mixture was observed to become cloudy, indicating that salt formation was occurring. After standing (as in Example 1), the sample was filtered and dried under vacuum.

[0137] Example 5: 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylate potassium salt.

[0138] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and potassium hydroxide were prepared using a procedure similar to that described in Example 4.

[0139] Example 6: 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and diethylamine.

[0140] Salts of 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and diethylamine were prepared using a procedure similar to that described in Example 4.

[0141] Example 7: 4-(2-((4-bromophenyl)thio)acetamide)-1-phenethyl-1H-pyrazole-3-carboxylic acid and diethylamine.

[0142] Before adding 480 mL of DIPE, approximately 8 g of free acid of compound 1 was weighed into a 500 mL glass container. To the resulting suspension, 1.1 equivalents of diethylamine, prepared as a 1 M stock solution in DIPE, were added. Upon addition of the base, precipitation of the sample from the reaction mixture was observed, indicating that salt formation had occurred. The sample was then allowed to stand for 4 days (8-hour cycles of 4 hours at 50°C and 4 hours at room temperature). After standing, the sample was filtered and dried under vacuum at 40°C for 2 days.

[0143] Example 8: Solubility in various solvents

[0144] The solvent is 100-250 μL Up to 1750 in increments μL Before increasing the amount to the maximum, approximately 10 mg of free acid of compound 1 was weighed into a 2 mL HPLC vial. After each addition of solvent, the sample was briefly shaken to promote dissolution and visually inspected for any signs of residual solid. Once the maximum amount of solvent was added, 1.1 equivalents of 1 M sodium hydroxide (aqueous solution) were also added to indicate which solvent yielded the appropriate salt form. The samples were visually inspected before standing for 2 days, and then all were inspected again. All samples were then evaporated at room temperature to produce solid material.

[0145] Analysis of the recovered solids by XRPD after standing showed the presence of a diffraction pattern (Pattern 1) frequently observed in sodium salts prepared from most of the solvents tested. However, four additional patterns were also identified, though less common. Salts exhibiting these five patterns were further analyzed by 1H NMR, from which the following conclusions were drawn. Pattern 1: Crystalline sodium salt (from most of the solvents tested) Pattern 2: Solvates of sodium salts (from 2-butanol and 2-propanol) Pattern 3: Possible new polymorphs (from 4-methylpentan-2-one) Pattern 4: Sodium salt containing unknown contaminants (from acetone) Pattern 5: Hemiethanolate of sodium salt (from ethanol).

[0146] The following list of peaks can be used to distinguish one of these patterns from another. The 2-theta values ​​are rounded to two decimal places.

[0147] Pattern 1 is characterized by 2-theta peaks at 4.68, 6.54, 9.24, 10.30, 13.80, 14.54, 16.60, 18.48, 18.96, 20.60, 22.18, 23.04, 23.49, 24.83, 25.76, 26.15, 26.97, 27.72, 28.09, 28.91, 29.63, 30.71, 31.03, 31.41, 32.10, 32.45, 32.75, 33.48, 33.76, and 34.74.

[0148] Pattern 2 is characterized by two-theta peaks at 7.00, 11.61, 15.75, 19.19, 20.30, 20.86, 23.19, 26.08, 26.72, and 29.29.

[0149] Pattern 3 is characterized by 2-theta peaks at 6.71, 9.47, 10.59, 13.13, 14.06, 14.86, 16.30, 16.90, 17.64, 18.81, 19.28, 20.92, 22.49, 23.39, 23.80, 24.73, 25.12, 26.05, 26.45, 27.27, 27.66, 28.35, 28.79, 29.20, 29.92, 31.00, 32.46, 34.01, and 35.09.

[0150] Pattern 4 is characterized by 2-theta peaks at 3.76, 6.47, 7.46, 8.28, 8.63, 11.34, 14.49, 15.78, 18.96, 19.27, 19.97, 21.64, 22.16, 23.24, 25.67, 27.61, 29.77, and 33.27.

[0151] Pattern 5 is characterized by two-theta peaks at 5.30, 5.83, 7.09, 10.57, 10.97, 11.75, 13.02, 13.80, 17.38, 17.97, 18.23, 18.75, 21.26, 22.10, 23.13, 23.50, 25.17, 26.84, 27.39, 28.76, 29.14, 29.57, 30.05, 31.18, 32.15, 33.44, 35.06, 36.29, and 39.26.

[0152] Table 2 shows the results of the solubility screening.

[0153] [Table 3] JPEG0007834307000012.jpg171158 JPEG0007834307000013.jpg119158

[0154] Example 9: Powder X-ray diffraction (XRPD) analysis

[0155] Samples were prepared by coating them onto a sample holder fitted with a 0 background silicon wafer (5 1 0). Analysis was performed using a Cu-Ka X-ray source operating at 40k, V40mA and LynxEye. TM The analysis was performed using a detector. All samples were analyzed over a range of 2 to 40°²θ.

[0156] XRPD analysis showed that the salt produced a crystalline solid with a novel crystalline pattern compared to free acid compound 1, and furthermore, the sample gave a partial crystalline pattern (see Table 3 and Figures 1-19 and 44).

[0157] [Table 4] JPEG0007834307000015.jpg91152

[0158] Example 10: Water-soluble

[0159] Water solubility was evaluated using approximately 5 mg of each solid that showed a unique pattern by XRPD. Before visual inspection, these samples were added to a fixed volume of deionized water up to a maximum of 4000 pL, and the samples were shaken during the addition process to promote dissolution. At room temperature, none of the samples were observed to dissolve, but after shaking overnight at 50°C, five samples were observed to be completely dissolved, and seven samples were partially dissolved.

[0160] Approximately 50 mg of each salt was weighed into a 2 mL HPLC vial before adding 1 mL of deionized water. The samples were then shaken at 25°C for 24 hours, dried overnight under vacuum, and weighed again before filtering into a pre-weighed filter cartridge. Solubility was then calculated. The experiment was repeated at 50°C (see Table 4).

[0161] [Table 5]

[0162] Example 11: Proton nuclear magnetic resonance spectroscopy (NMR)

[0163] Samples for NMR analysis were prepared by weighing 5–7 mg of the sample into a 1.5 mL HPLC vial before dissolving in d-DMSO, and then transferring the sample to a 5 mm NMR tube suitable for analysis. The samples were analyzed using standard instrument settings.

[0164] ¹H NMR data of fully soluble or partially soluble salts prepared during screening showed that they all had different chemical shifts for the proton-related peaks around the carboxylate group compared to those of the free acid. This indicates salt formation (the peaks at 9.91, 8.22, 4.38, and 4.03 of free acid compound 1 show the most significant changes in the shift; see Table 5). Some samples also showed the solvent present in the NMR, which may result from either insufficient drying or solvate formation; water was also observed in all NMR data, which may be related to the preparation. Where counterions were visualized by ¹H NMR, they were also quantified (see Table 5).

[0165] [Table 6]

[0166] Example 12: Differential Scanning Calorimetry (DSC)

[0167] Approximately 1–3 mg of the sample was placed on a pre-weighed aluminum DSC dish using an analytical balance. The sample was heated at 10°C / min under a nitrogen atmosphere from room temperature to a temperature approximately 5°C above the decomposition point. Each dataset was examined for thermal phenomena.

[0168] Example 13: Fourier transform infrared spectroscopy (FT-IR)

[0169] Approximately 1-2 mg of the sample was placed on the crystal of the ATR module and fixed in place. All generated data was corrected by removing background noise in the analysis software.

[0170] Example 14: Polarized Light Microscopy (PLM)

[0171] Samples were prepared on glass microscope slides using 1-2 drops of immersion oil and a glass coverslip. Optical evaluation of the samples was performed using appropriate objective lenses with polarizers in the crossing, partially crossing, and non-crossing positions.

[0172] Example 15: Hot Stage Microscope (HSM)

[0173] The sample was prepared on a glass microscope slide, and the temperature profile used in TGA and DSC was mimicked. The sample was heated to its melting point at 10°C / min, and then cooled to room temperature without forced cooling.

[0174] Example 16: Dynamic water vapor adsorption (DVS)

[0175] Approximately 10–15 mg of sample was weighed into a stainless steel DVS basket before submission for analysis. Samples were analyzed for up to 6 hours at each humidity level in the range of 0–90% RH. Each sample was subjected to two cycles. XRPD analysis was performed on all samples after DVS.

[0176] Example 17: Thermogravimetric Analysis (TGA)

[0177] The samples were heated from room temperature to 400°C at a rate of 10°C / min (unless otherwise specified) under a nitrogen gas stream. The mass loss and decomposition temperature of each sample were determined by examining the data sets.

[0178] Example 18: Inductively coupled plasma (ICP)

[0179] Approximately 0.10 g of the test sample was pulverized with 5 mL of nitric acid and then diluted to the final volume with deionized water. The test sample was then further diluted and analyzed against a series of calibration standards to determine the sodium and potassium content.

[0180] Example 19: Karl Fischer

[0181] Approximately 0.05g of the test sample was returned to the KF container and weighed, then Hydranal (R) -The salt's % water content was determined by titration with Composite 5.

[0182] Example 20: Particle size

[0183] Dispersant: air, Lens: R3 (potassium and diethylamine) and R5 (sodium), Pressure: 4 bar, Feeding rate: 40 mm / s, Optical model: Fraunhofer, Measurement time: 5 seconds. Two parts of the sample were analyzed as dry powder, and the average of the recorded values ​​was reported.

[0184] Example 21: HPLC

[0185] Flow rate: 3.03 mL / min, Method: Isocratic, Column temperature: 25°C, Wavelength range: 190-400 nm, Solvent A: 25 mM ammonium acetate buffer - pH 5.5 (30%), Solvent B: MeOH (70%), Injection volume: 15 μL, Run time: 20 minutes.

[0186] Example 22: Artificial intestinal fluid in a fasted state / Artificial intestinal fluid during feeding / Solubility

[0187] Approximately 25 mg of each salt was weighed into a 2 mL HPLC vial before adding 1 mL of fasting intestinal fluid (FaSSIS) solution. The samples were then shaken at 37°C for 24 hours, dried overnight under vacuum, and weighed again, from which the solubility was calculated, before filtering into a pre-weighed filter cartridge. The experiment was also repeated using intestinal fluid (FeSSIF) solution during feeding.

[0188] Example 23: pH 1 stable

[0189] Approximately 25 mg of each salt was weighed into a 2 mL HPLC vial before adding 1 mL of pH 1 buffer. The samples were then shaken at 37°C for 4 hours and dried overnight before filtering into an SPE cartridge.

[0190] Example 24: pH

[0191] Before analysis, saturated solutions of each salt were prepared in 5 mL of deionized water at room temperature.

[0192] [Table 7] JPEG0007834307000019.jpg222150 JPEG0007834307000020.jpg89150

[0193] [Table 8] JPEG0007834307000022.jpg195151

[0194] [Table 9] JPEG0007834307000024.jpg215150 JPEG0007834307000025.jpg61150

[0195] <Pharmaceutical Products>

[0196] Since the disclosed salt of Formula 2 is not stable under acidic conditions, formulations for clinical use should be prepared with appropriate buffering and / or coatings so that they can survive under gastric conditions (e.g., "enteric-coated" formulations) or be administered via routes other than oral (e.g., by injection or patch).

[0197] Preparation of salts of Formula 2 in dosage forms for oral administration, injection, transdermal patch administration, and those containing excipients such as flavoring agents, buffers, and carriers, and packaging of dosage forms, are considered to be within the scope of the art. For example, see Remington: the Science and Practice of Pharmacy, 22nd Ed., c. 2013 by Pharmaceutical Press, which is incorporated herein by reference in its entirety for all purposes. Formulations should be prepared and administered to provide subjects with doses in the range of 1 to 1000 mg / day, or 1 to 100 mg / day, or 10 to 100 mg / day.

Claims

1. Formula 2: 【Chemistry 1】 [In the formula, n is 1, m is 1.5, X ＋ is a potassium ion, and Sol is a water molecule. A salt compound containing [sol].

2. Formula 2: 【Chemistry 2】 [In the formula, n is 1, m is 2, X+ is a sodium ion, and Sol is a water molecule. A salt compound containing [sol].

3. Formula 2: 【Transformation 3】 [In the formula, n is 1, m is 0, X+ is the cation of diethylamine, and Sol is a water molecule. A salt compound containing [sol].

4. A method for preparing a salt compound according to any one of claims 1 to 3, i) The following compound 1 is added to the organic solvent selected from the group consisting of ethanol, 4-methylpentan-2-one, diisopropyl ether (DIPE), or a mixture of any two or three thereof, or to the organic solvent mixed with water: Dissolving; ii) Adding an excess amount of base in a quantity exceeding the stoichiometric amount required to titrate compound 1 to form a precipitate of the salt compound of formula 2, where the base is a salt of potassium hydroxide, sodium hydroxide, or diethylamine cation; and iii) Collect the precipitate to obtain the salt compound. Methods that include...

5. The method according to claim 4, wherein the organic solvent is ethanol or diisopropyl ether (DIPE).

6. The method according to claim 4 or 5, wherein the organic solvent mixed with water is a polar organic solvent mixed with water.

7. The method according to claim 6, wherein the ratio of water to a polar organic solvent is in the range of 5:1 to 10:0.1.

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