Salt forms of organic compound

Abstract

To provide a novel salt form of 4-(2-((4-bromophenyl)thio)acetamido)-1-phenethyl-1H-pyrazole-3-carboxylic acid and a method for producing the same.SOLUTION: The present invention provides a salt compound having formula 2, wherein X+ is a potassium ion, a sodium ion, or a quaternary dimethylamine or quaternary diethylamine.SELECTED DRAWING: None

Description

[Technical Field]

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

[0002] Ischemia refers to a reduction in blood flow to an organ, tissue, or region caused by the constriction or blockage of one or more blood vessels. Once ischemia occurs, various sequelae develop due to neuronal damage, even if reperfusion is rapid. Such ischemia frequently occurs in coronary artery disease, cardiovascular disease, angina, headaches, and other conditions associated with vascular blockage or constriction, ultimately resulting in irreversible damage, i.e., cell or tissue necrosis.

[0003] Ischemic diseases, such as myocardial infarction, arrhythmias, and heart failure, which result from cellular damage and dysfunction during ischemia-reperfusion, have been the subject of ongoing basic and clinical research for the past 50 years due to their high morbidity, mortality, and low complete cure rates [Wang, QD et al., Cardiovasc. Res. 55:25-37, 2002]. In particular, ischemia-reperfusion injury is associated with various physiological mechanisms, including alterations in metabolism, immune responses, ion homeostasis, and oxygen free radical generation. Therefore, research has been conducted in various fields related to immunomodulation, cell death prevention, ion channel regulation, and other areas [Hearse, DJ et al., Mol. Cell. Biochem. 186:177-184, 1998]. Based on such mechanistic studies, numerous therapeutics and surgical procedures focusing on novel sites of action have been developed, but no commercially available technology has yet been developed to protect cardiomyocytes from ischemia-reperfusion injury. Therefore, there is a need for drugs for preventing and treating ischemic heart disease or drugs for preventing heart disease that can slow the progression of ischemic damage to cardiomyocytes and reduce reperfusion-induced damage.

[0004] Furthermore, it has been shown that the alleviation of ischemia through the restoration of blood flow promotes the production of reactive oxygen species (ROS), causing a significant decrease in glutathione and resulting in more severe disease. Similar disorders are observed when blood flow is stopped or restored during various techniques in transplant surgery for various types of organs, such as the heart, liver, lung, pancreas, or blood vessels, and are also problematic during organ dissection and removal. Reactive oxygen species and reactive free radicals, which are suspected to cause disease, are detected in the cytoplasm and organelles of tissue cells, particularly mitochondria, which produce ATP as the primary energy source for cells. In mitochondria, these reactive molecules are primarily released 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 cell necrosis, and cell death that occurs especially after reperfusion is a major cause of tissue damage, and 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] During cerebral ischemia, the depletion of energy sources due to a reduced blood supply induces ischemic cell death, which then overactivates cell membrane receptors, leading to various biochemical changes inside and outside the cells, including accumulation of glutamate and calcium, and damage to lipids, proteins, and nucleic acids, ultimately resulting in brain tissue damage (Liu, P.K., 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 arrhythmia, ischemic cell death has been reported to occur through the activation of lipid enzymes that cause damage to cell membranes and subsequent 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]. It is known that glutamate-mediated cell death of retinal cells during retinal ischemia is 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 subsequent arterial occlusive damage due to cell necrosis and hemodynamic disturbance 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 in 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 trauma, and neonatal hypoxia [GJ Zoppo et al., Drugs 54, 9 (1997); I. Sziraki et al., Neurosci. 85, 1101 (1998)]. Summary of the Invention [Means for solving the problem]

[0010] Disclosed are salt compounds having formula (2) (below).

[0011] [ka]

[0012] Formula (2): [n is 1, 2, or 3, m may be a non-integer number between 0 and 3, and m is typically 0, 0.5, 1, 2, or 3; "Sol" is a solvent molecule, 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.

[0013] Salt compounds of Formula 2 can be prepared by treating the free base or zwitterion of Compound 1 (described below) 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 drawings]

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

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

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

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

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

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

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

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

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

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

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

[0025] [Figure 12] Figure 12 shows XRPD diffraction patterns prepared with N-ethylglucamine in various solvents: 1 from 4-methyl-pentan-2-one, 2 from diisopropyl ether, 3 from ethanol, and 4 from the free acid.

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

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

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

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

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

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

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

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

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

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

[0036] [Figures 23A-E] Figures 23A-23E show HSM analysis of potassium salt Formula 2 from a second preparation of KM-819. 23A: Initial condition of potassium salt, 23B: Loss of birefringence at 127°C, 23C: Initial melt at 154°C, 23D: Secondary melt at 212°C, 23E: Recrystallization.

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

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

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

[0040] [Figure 27] FIG. 27 shows the DVS isotherm plot of potassium salt Formula 2 from a second preparation of KM-819.

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

[0042] [Figure 29A-C] 29A-29C show HSM analysis of the sodium salt from a second preparation of KM-819. 29A: As-prepared salt, 29B: Melting at 136° C., 29C: Recrystallization.

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

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

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

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

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

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

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

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

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

[0052] [Figure 39A-E] Figures 39A-E show HSM analysis of the diethylamine salt Formula 2 from a second preparation of KM-819. 39A: As-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] FIG. 40 shows an FT-IR analysis of the diethylamine salt Formula 2 from a second preparation of KM-819.

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

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

[0056] [Figure 43] FIG. 43 shows the DVS isotherm plot of the diethylamine salt Formula 2 from a second preparation of KM-819.

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

[0058] [Figure 45] Figure 45 shows an alignment of five different H NMR analyses observed for samples from the salt screen. The spectra are aligned through the DMSO standard peak. The pattern numbering corresponds to the XRPD pattern numbering, and salts that show XRPD pattern 1 often show NMR pattern 1. DETAILED DESCRIPTION OF THE INVENTION

[0059] Aminopyrazole derivatives have been disclosed to inhibit ischemic cell death and can therefore be used as agents for preventing and treating 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, head trauma, Alzheimer's disease, Parkinson's disease, neonatal hypoxia, glaucoma, and diabetic neuropathy, and for protecting organs during transplant surgery.

[0060] Compound 1 (KM-819) is a novel aminopyrazole derivative useful for the treatment of Parkinson's disease. The disclosed compound 1 (KM-819) can be synthesized as described in WO2008 / 051047 (incorporated herein by reference in its entirety for all purposes) to provide a white crystalline powder. Initial analysis of compound 1 (KM-819) as the free acid or zwitterion was performed to better understand the material and to provide baseline data so that comparisons could be made between compound 1 (KM-819) and any salts prepared.

[0061] Those skilled in the art will appreciate 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 the zwitterionic form of the compound, depending on the pH of a solution of Compound 1.

[0062] [ka]

[0063] A solubility evaluation of compound 1 (KM-00819) was performed using water and a range of organic solvents. A list of solvents suitable for use during salt screening was determined from this evaluation. Upon completion of the solvent screening, 1.1 equivalents of sodium hydroxide (aqueous) was added to each sample to test the sample's ability to generate a salt form. Upon hydroxide addition, a cloudy precipitate formed in some samples, indicating that salt formation had likely occurred, and five distinct diffraction patterns were observed in XRPD analysis of solids isolated from salt formation. (See, e.g., Example 8 and Figures 10-19 and 44.) H NMR analysis of samples exhibiting diffraction patterns was used to determine the most suitable solvents for use in preparing salt forms of KM-819, as described below. Figure 45 shows an alignment of representative H NMR analyses of samples from the salt screening.

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

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

[0066] Disclosed herein are novel pharmaceutically acceptable solid forms of KM-819 and salts thereof and methods for their preparation. These forms can be used to prepare salts or bases and to prepare formulations thereof for clinical use.

[0067] Disclosed herein are novel crystalline or amorphous forms of 4-(2-((4-bromophenyl)thio)acetamido)-1-phenethyl-1H-pyrazole-3-carboxylic acid (KM-819) pharmaceutically acceptable salts and methods for their preparation. These salts can be used to prepare other salts or the free base form 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 formula 2 below:

[0069] [ka]

[0070] In Formula 2, n is an integer of 1, 2, or 3; m is between 0 and 3, and may be a non-integer, such as 0.5 or 1.5, "Sol" is a solvent molecule, 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.

[0071] 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 salt compounds of Formula 2, m may be 0.5 or 1. In such examples, X + may be a potassium ion, a sodium ion or a quaternary methylamine or a quaternary ethylamine.

[0073] Salt Compound 2 may alternatively or additionally be in the form of a solvate containing water, ethanol, or diisopropyl ether, or a mixture of any two or three thereof. The solvent molecules may be present in a non-integer ratio relative to either or both of the water molecules and Compound 1 ion, for example, 0.1, 0.2, or 0.5 solvent molecules per molecule of Compound 1 ion. The solvent molecules may be present in an integer ratio relative to either or both of the water molecules and Compound 1 ion, for example, 1 or 2 solvent molecules per molecule of Compound 1 ion.

[0074] Also, equation (2):

[0075] [ka]

[0076] [In the formula, n is 1, 2, or 3; m is 0 to 3; X + is a cation, "Sol" is a solvent molecule. i) dissolving 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 of base in excess of 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) Collecting the precipitate to obtain salt compound 2 A method is also disclosed, including:

[0077] In such methods, 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-methyl-pentan-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 quaternary dimethylamine, or a salt of quaternary diethylamine.

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

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

[0082] The water may be mixed with an organic solvent, and in some cases, the water may be mixed with ethanol or DIPE.

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

[0084] The free base (zwitterion) Compound 1 may be dissolved in unbuffered water, a range of organic solvents, mixtures of organic solvents, and mixtures of solvents with 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-methyl-pentan-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. Example solubility results are shown in Table 2 below.

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

[0086] For binary mixtures, the ratio of water to organic solvent (preferably a polar organic solvent) in the solvent mixture may be from 1:10 to 1:0.1, or from 1:5 to 1:0.1, or from 1:2-1:0.1, or from 1:2-1:0.5, or about 1:1.

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

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

[0089] [ka]

[0090] The free acid Compound 1 is weighed and added to a vessel, followed by the addition of solvent. Approximately 1.1 equivalents of base, prepared as a 1 M stock solution in the solvent, is then added. Cloudiness of the sample upon addition of the base indicates salt formation. After standing (left at room temperature for several hours), the sample is filtered, dried under vacuum, and then characterized by various methods.

[0091] The base 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 include amorphous phases (disordered), hydrates (water present in the crystal lattice), and solvates (solvents other than water present in the crystal lattice).

[0094] Since different crystalline modifications have different crystalline structures and different free energies, polymorphs exhibit different physico-chemical properties such as melting point, density, solubility, chemical stability and ultimately bioavailability.

[0095] An example of a preferred salt of compound 2, 4-(2-((4-bromophenyl)thio)acetamido)-1-phenethyl-1H-pyrazole-3-carboxylic acid, is as follows:

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

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

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

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

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

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

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

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

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

[0105] Salt of 4-(2-((4-bromophenyl)thio)acetamido)-1-phenethyl-1H-pyrazole-3-carboxylic acid and N-(phenylmethyl)benzeneethanamine,

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

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

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

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

[0110] Many of the solids isolated from the salt formation step exhibited diffraction patterns in XRPD analysis that indicated the solids were crystalline, and some exhibited diffraction patterns that differed from that of the 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 use in forming the salt is one that completely dissolves the free acid Compound 1, and a suitable base for use in preparing the salt Compound 2 from Compound 1 is one that completely replaces the carboxylic acid hydrogen of Compound 1.

[0112] [Table 1] [Example]

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

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

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

[0116] <Equipment used> Perkin Elmer PYRIS 1 DSC using 40 μL aluminum pans (vented). Data collection and analysis were performed using Perkin Elmer control and analysis software, version 11.0.2.0468.

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

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

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

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

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

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

[0123] SMS DVS-specific dynamic vapor sorption apparatus 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 implemented in Microsoft Excel. Analysis was performed as a wt% change from 0 to 90% RH, and isothermal plots were also examined.

[0124] Perkin Elmer TGA PYRIS 1 using an aluminum dish (vented) in a ceramic crucible. Data analysis and collection were performed using Perkin Elmer control and analysis software, version 11.0.2.0468.

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

[0126] The Metrohm 852 Titranto is a combined volumetric and coulometric KF unit. All samples were analyzed using the volumetric Karl Fischer module.

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

[0128] Heidolph Titramax 1000 with heating module.

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

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

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

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

[0133] In the same manner as in Example 1 or Example 2, salts of the following compounds were prepared.

[0134] [Table 2] JPEG2025131638000008.jpg205152 JPEG2025131638000009.jpg212152 JPEG2025131638000010.jpg130153

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

[0136] Approximately 1 g of Compound 1 was weighed into 3 x 100 mL vials before 60 mL of DIPE was added to each container. To the resulting suspension was added 1.1 equivalents of sodium hydroxide, prepared as a 1 M stock solution in water. Upon addition of the base, the reaction mixture was observed to become cloudy, indicating salt formation had occurred. After settling (as in Example 1), the samples were filtered and dried under vacuum.

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

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

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

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

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

[0142] Approximately 8 g of the free acid of Compound 1 was weighed into a 500 mL glass container before adding 480 mL of DIPE. To the resulting suspension, 1.1 equivalents of diethylamine, prepared as a 1 M stock solution in DIPE, was added. Upon addition of the base, the sample was observed to precipitate from the reaction mixture, indicating salt formation. 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] Approximately 10 mg of the free acid of compound 1 was weighed into a 2 mL HPLC vial before adding solvent in 100-250 μL increments up to 1750 μL. After each solvent addition, the sample was briefly shaken to facilitate dissolution and visually inspected for signs of residual solids. Once the maximum amount of solvent had been added, 1.1 equivalents of 1 M sodium hydroxide (aqueous) solution was also added, providing an indication as to which solvent would yield the appropriate salt form. The samples were visually inspected before standing for two days, and then all were inspected again. All samples were then allowed to evaporate at room temperature to produce solid material.

[0145] Analysis of the recovered solid by XRPD after standing showed a frequently observed diffraction pattern (pattern 1) observed for the sodium salt prepared from most solvents tested. However, four additional patterns, although less common, were also identified. 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 polymorph (from 4-methylpentan-2-one) Pattern 4: Sodium salt (from acetone) containing unknown contaminants Pattern 5: Sodium salt hemiethanoate (from ethanol).

[0146] The following list of peaks can be used to distinguish one of these patterns from another, with 2-theta values ​​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 peaks at 2-theta of 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 peaks at 2-theta of 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 2-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] JPEG2025131638000012.jpg171158 JPEG2025131638000013.jpg119158

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

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

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

[0157] [Table 4] JPEG2025131638000015.jpg91152

[0158] Example 10: Water Soluble

[0159] Water solubility assessments were performed using approximately 5 mg of each solid that displayed a unique pattern by XRPD. Prior to visual inspection, these samples were added to aliquots of deionized water up to 4000 μL, with the samples being shaken between additions to promote dissolution. None of the samples were observed to dissolve at room temperature, but after overnight shaking at 50°C, five samples were observed to dissolve completely, and an additional 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 before being filtered onto a pre-weighed filter cartridge, dried overnight under vacuum, and reweighed, from which the solubility was calculated. The experiment was also 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 sample into a 1.5 mL HPLC vial before dissolving in d-DMSO. The sample was then transferred to a 5 mm NMR tube prepared for analysis. Sample analysis was performed using standard instrument settings.

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

[0165] [Table 6]

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

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

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

[0169] Approximately 1–2 mg of sample was placed on the crystal of the ATR module and fixed in place. All generated data were corrected by background subtraction 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 objectives with polarizers in crossed, partially crossed, and uncrossed positions.

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

[0173] Samples were prepared on glass microscope slides and heated at 10°C / min to the melting point of the sample, mimicking the temperature profile used in TGA and DSC, and then cooled to room temperature without forced cooling.

[0174] Example 16: Dynamic Vapor Sorption (DVS)

[0175] Approximately 10-15 mg of sample was weighed into a stainless steel DVS basket before being submitted for analysis. Samples were analyzed at 0-90% RH for up to 6 hours per humidity step. Each sample was cycled twice. XRPD analysis of all samples was performed after DVS.

[0176] Example 17: Thermogravimetric Analysis (TGA)

[0177] The samples were heated under a stream of nitrogen gas at 10°C / min (unless otherwise indicated) from room temperature to 400°C. Each data set was examined to determine the mass loss and decomposition temperature of the sample.

[0178] Example 18: Inductively Coupled Plasma (ICP)

[0179] Approximately 0.10 g of test sample was digested with 5 mL of nitric acid and made to 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 0.05 g of test sample was weighed back into the KF container and (R) -Titrated with Composite 5 to determine the % water content of the salt.

[0182] Example 20: Particle size

[0183] Dispersant: air, Lenses: R3 (potassium and diethylamine) and R5 (sodium), Pressure: 4 bar, Feed rate: 40 mm / s, Optical model: Fraunhofer, Measurement time: 5 seconds, Samples were analyzed in duplicate as dry powders 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 min.

[0186] Example 22: Simulated intestinal fluid in fasting state / simulated intestinal fluid in fed state / solubility

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

[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 allowed to dry overnight before filtering onto an SPE cartridge.

[0190] Example 24: pH

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

[0192] [Table 7] JPEG2025131638000019.jpg222150 JPEG2025131638000020.jpg89150

[0193] [Table 8] JPEG2025131638000022.jpg195151

[0194] [Table 9] JPEG2025131638000024.jpg215150 JPEG2025131638000025.jpg61150

[0195] <Formulation>

[0196] Because the disclosed salts of Formula 2 are not stable under acidic conditions, formulations for clinical use should be prepared with appropriate buffers and / or coatings to survive gastric conditions (e.g., "enteric" formulations) or to be administered by other than oral routes (e.g., by injection or patch).

[0197] The preparation of salts of Formula 2 in dosage forms for oral administration, injection, transdermal patch administration, and the like, and packaging the dosage forms with excipients such as flavorings, buffers, carriers, etc., is believed to be within the skill of one of ordinary skill in the art. See, e.g., Remington: The Science and Practice of Pharmacy, 22nd Ed., c. 2013 by Pharmaceutical Press, incorporated herein by reference in its entirety for all purposes. Formulations should be prepared and administered to provide a subject with a dose ranging from 1 to 1000 mg / day, or to provide a subject with a dose ranging from 1 to 100 mg / day, or to provide a subject with a dose ranging from 10 to 100 mg / day.

Claims

1. Formula (2): 【Chemical 1】 [In the formula, n is 1, 2 or 3; m is 0, 0.5, 1, 2, or 3; X ＋ is a cation, and Sol is a solvent molecule. A salt compound having the formula:

2. X ＋ 2. The salt compound of claim 1, wherein is a potassium ion, a sodium ion, a calcium ion, or a substituted ammonia molecule.

3. X ＋ is a quaternary amine selected from the group consisting of choline, diethylamine, dimethylamine, L-lysine, N,N'-dibenzylethylenediamine, N-ethylglucamine, 1-(2-hydroxyethyl)pyrrolidine, N-(phenylmethyl)benzeneethanamine, ammonia, magnesium acetate, N-methylglucamine, tromethamine, 4-(2-hydroxyethyl)morpholine, 2-(diethylamino)ethanol, and 2-dimethylaminoethanol.

4. X ＋ 2. The salt compound of claim 1, wherein is a potassium ion, a sodium ion, or a quaternary dimethylamine or a quaternary diethylamine.

5. The salt compound of claim 1, wherein m is 0.5 or 1.

6. The salt compound according to claim 4, wherein m is 0.5 or 1.

7. The salt compound of claim 1, which is a solvate further containing 4-methyl-pentan-2-one, ethanol, or diisopropyl ether.

8. The salt compound of claim 4, which is a solvate further containing ethanol or diisopropyl ether.

9. Formula (2): 【Chemistry 2】 [In the formula, n is 1, 2 or 3; m is 0, 0.5, 1, 2, or 3; X ＋ is a cation, and wherein Sol is a solvent molecule. i) dissolving 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 of base in excess of 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) Collecting the precipitate to obtain salt compound 2 A method comprising:

10. 10. The method of claim 9, wherein the organic solvent is 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 thereof.

11. 10. The method of claim 9, wherein the organic solvent is ethanol or diisopropyl ether (DIPE).

12. 10. The method of claim 9, wherein the base is sodium hydroxide, potassium hydroxide, or a salt of a quaternary dimethylamine or a salt of a quaternary diethylamine.

13. 12. The method of claim 11, wherein the base is sodium hydroxide, potassium hydroxide, or a salt of a quaternary dimethylamine or a salt of a quaternary dimethylamine.

14. 10. The method of claim 9, wherein the solvent is water mixed with a polar organic solvent.

15. 15. The method of claim 14, wherein the ratio of water to polar organic solvent ranges from 5:1 to 10:0.

1.

16. 12. The method of claim 11, wherein the solvent is water mixed with a polar organic solvent.

17. 17. The method of claim 16, wherein the ratio of water to polar organic solvent ranges from 5:1 to 10:0.

1.

18. 13. The method of claim 12, wherein the solvent is water mixed with a polar organic solvent.

19. 20. The method of claim 18, wherein the ratio of water to polar organic solvent ranges from 5:1 to 10:0.1.

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