KR20210026308A - A salt and crystalline form of a benzopyranone compound, a preparation method thereof, and a pharmaceutical composition comprising the same - Google Patents

Abstract

A salt and a crystal form of a compound in chemical formula 1, namely 2-(2,5-difluorophenyl)-3- carboxy-7-chloro-(4H)-4-benzopyranone, according to the present invention are excellent in terms of various physicochemical properties, which are hygroscopicity, physicochemical stability, solubility and the like, and thus can be utilized for manufacturing a pharmaceutical composition comprising the same as an active component.

Description

Salt and crystal form of a benzopyranone compound, a method for preparing the same, and a pharmaceutical composition comprising the same TECHNICAL FIELD [A SALT AND CRYSTALLINE FORM OF A BENZOPYRANONE COMPOUND, A PREPARATION METHOD THEREOF, AND A PHARMACEUTICAL COMPOSITION COMPRISING THE SAME}

Salts and crystal forms of 2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone according to the present invention, a method for preparing the same, and a pharmaceutical composition comprising the same About.

TNF is a type of cytokine, and normal TNF plays an important role in the inflammatory response. However, overproduced TNF stimulates macrophages to induce excessive inflammatory reactions in the cells, resulting in rheumatoid arthritis, inflammatory bowel disease, psoriasis, and ankylosing spondylitis. Causes TNF-related diseases. As TNF inhibitors for treating TNF-related diseases, etanercept (Enbrel®), adalimumab (Humira®), infliximab (Remicade®), and the like are commercially available. Etanercept is a receptor-based preparation, which is a fusion protein preparation of an IgG fixed site (Fc) and a human TNF receptor, and adalimumab and infliximab are antibody-based preparations, respectively, which are human recombinant IgG monoclonal antibodies and chimeric antibodies. Since etanercept contains the TNF receptor, it first binds only with TNF floating in the blood before it reaches the receptor on the cell membrane and acts to prevent it from sending inflammatory signals.Adalimumab and infliximab act as well as free-type TNF floating in the blood. , It is characterized by binding both to the adherent TNF attached to the TNF receptor on the cell membrane.

The biopharmaceuticals of the TNF inhibitor show excellent efficacy in a short period of time, but are expensive and require repetitive injections, so patients' reluctance is high, and about 1/3 of the patients have no therapeutic effect, and patients who respond to the drug are also Resistance develops within several years due to immunogenic side effects, and it has distinct disadvantages such as difficult storage because low temperature storage is essential, and it has unmet medical demand.

Korean Patent Registration No. 10-1934651 discloses a 4-benzopyranone derivative as a novel compound that directly binds to TNF and inhibits TNF activity. In particular, according to Korean Patent Registration No. 10-1934651, 2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzo, which is a compound having the structure of Formula 1 below. Pyranone (hereinafter, "the compound of Formula 1") has excellent cytotoxic inhibitory effect of TNF and cell binding inhibitory effect of TNF, inhibits cell signaling by TNF, and induces sepsis, rheumatoid arthritis, inflammatory bowel disease, sepsis. It shows therapeutic effects such as acute kidney injury, and shows comparable effects even when compared to commercially available adalimumab and etanercept.

[Formula 1]

In order for a drug to exhibit high pharmacological activity in vivo, the drug must be rapidly eluted from the digestive tract. This is closely related to the solubility of the drug. As the solubility of the drug increases, the dissolution rate of the drug and the absorption rate of the digestive tract increase, and the faster the absorption rate in the digestive tract, the higher the blood concentration in the blood even at a low dose. You can expect a high effect. Therefore, it is required to select the optimal salt form, and physical and chemical properties such as solubility, stability, hygroscopicity, processability into tablet formulations, purity, or density must be considered in order to prepare a salt with excellent physical properties. .

Korean Patent Registration No. 10-1934651 Korean Patent Registration No. 10-1982667

It is an object of the present invention to provide a novel salt of a compound of Formula 1 and a crystalline form thereof of high purity, which is excellent in hygroscopicity, physicochemical stability, solubility, etc., can be mass-produced, and has a homogeneous quality.

Another object of the present invention is to provide a method for preparing a salt of the compound of Formula 1.

Another object of the present invention is to provide a pharmaceutical composition comprising the salt of the compound of Formula 1 as an active ingredient.

The present invention provides a compound of Formula 1 in the form of a salt selected from the group consisting of potassium salt, sodium salt, L-arginine salt, L-lysine salt, ammonia salt, meglumine salt and tromethamine salt.

[Formula 1]

In a preferred embodiment, the compound of Formula 1 may be in the form of a potassium salt, meglumine salt, or tromethamine salt. Most preferably, the present invention provides a potassium salt compound of formula 1.

In one embodiment, the potassium salt compound is X containing characteristic peaks at 4 or more diffraction angles 2θ ± 0.2° selected from the group consisting of 4.6°, 9.0°, 9.3°, 16.0°, 19.8°, and 31.7°. -May be a crystalline form showing a ray powder diffraction (XRPD) spectrum. In another embodiment, the potassium salt compound may have an endothermic peak in the range of 80°C to 120°C, preferably 87°C to 113°C when analyzed by a differential scanning calorimeter (DSC) under a temperature rising condition of 10°C/min. .

The present invention also comprises the steps of (a) adding a compound of formula 1 to ethyl acetate (EtOAc) to obtain a solution; (b) preparing a compound of Formula 1 in the form of a potassium salt by adding potassium hydroxide to the solution; And/or (c) crystallizing the compound of Formula 1 in the form of the potassium salt, providing a method for preparing the compound of Formula 1 in the form of a potassium salt.

The present invention also contains as an active ingredient of the compound of formula 1 in the form of potassium salt, rheumatoid arthritis, juvenile rheumatoid arthritis, inflammatory growth disease, Crohn's disease, ulcerative colitis, psoriasis, plaque psoriasis, juvenile plaque psoriasis, psoriatic arthritis, Polyarticular Pediatric Idiopathic Arthritis, Behcet's Enteritis, Ankylosing Spondylitis, Axial Spine Arthritis, Pediatric Bone Attachment Arthritis, Osteoarthritis, Multiple Rheumatoid Myalgia, Multiple Sclerosis, Systemic Lupus Erythematosus, Asthma, Sjogren's Syndrome, Pneumonia, Chronic Obstructive Pulmonary Disease, Sarcoidosis , Gori granuloma, Wegener's granulomatosis, arteriosclerosis, vasculitis, heart failure, myocardial infarction, kidney damage, nephritis, graft versus host disease, dementia, Alzheimer's disease, Parkinson's disease, pain, uveitis, Behcet's disease, purulent sweating, follicular red It provides a pharmaceutical composition for preventing or treating a TNF overexpression disease selected from the group consisting of nasal inflammation, diabetic milk fat necrosis production, gangrene pyoderma, sweet syndrome, subkeratinous pustular dermatosis, scleroderma, dermatomyositis, sepsis, and septic shock.

The salt and crystalline form of the compound of formula 1 according to the present invention, i.e. 2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H)-4-benzopyranone, has various physicochemical properties, namely It is excellent in terms of hygroscopicity, physicochemical stability, solubility, etc., and can be easily used in the manufacture of a pharmaceutical composition containing it as an active ingredient.

^{1 shows 1} HNMR results of various salts of the compound of Formula 1 prepared according to Example 1. FIG.
Figure 2 shows the XRPD pattern of various salts of the compound of Formula 1 prepared according to Example 1.
3 is a graph showing an example of an XRPD pattern of a potassium salt of a compound of Formula 1;
4 shows a PLM picture of the potassium salt of the compound of Formula 1.
5 is a graph showing an example of an endothermic peak and a TGA curve of the potassium salt of the compound of Formula 1 during DSC analysis.
6 is a graph showing DVS results of the potassium salt of the compound of Formula 1;
7 is a graph comparing the XRPD pattern of the potassium salt of the compound of Formula 1 with the XRPD pattern after DVS analysis thereof.
8 is a graph showing an example of the XRPD pattern of the sodium salt of the compound of Formula 1.
9 shows a PLM picture of the sodium salt of the compound of Formula 1.
10 and 11 are graphs showing examples of endothermic peaks and TGA curves appearing during DSC analysis of crystals of Form I and Form II of the sodium salt of the compound of Formula 1;
12 is a graph showing an example of the XRPD pattern of the L-arginine salt of the compound of Formula 1;
13 shows a PLM picture of the L-arginine salt of the compound of Formula 1.
14 is a graph showing an example of an endothermic peak and a TGA curve appearing during DSC analysis of the L-arginine salt of the compound of Formula 1;
15 is a graph showing an example of the XRPD pattern of the L-lysine salt of the compound of Formula 1;
16 shows a PLM picture of the L-lysine salt of the compound of Formula 1.
17 and 18 are graphs showing examples of endothermic peaks and TGA curves appearing during DSC analysis of I- and II-form crystals of the L-lysine salt of the compound of Formula 1;
19 is a graph showing an example of the XRPD pattern of the ammonia salt of the compound of Formula 1;
20 shows a PLM picture of the ammonia salt of the compound of Formula 1.
21 is a graph showing an example of an endothermic peak and a TGA curve appearing during DSC analysis of an ammonia salt of a compound of Formula 1;
22 is a graph showing an example of the XRPD pattern of the meglumine salt of the compound of Formula 1;
23 shows a PLM picture of the meglumine salt of the compound of Formula 1.
24 and 25 are graphs showing examples of endothermic peaks and TGA curves appearing during DSC analysis of crystals of Form I and Form II of meglumine salt of the compound of Formula 1;
26 shows an HSPLM picture of the meglumine salt of the compound of Formula 1.
FIG. 27 is a graph showing DVS results of Form I crystals of meglumine salt of the compound of Formula 1. FIG.
28 is a graph comparing the XRPD pattern of the Form I crystal of the meglumine salt of the compound of Formula 1 with the XRPD pattern after DVS analysis thereof.
29 is a graph showing DVS results of Form II crystals of meglumine salt of the compound of Formula 1;
30 is a graph comparing the XRPD pattern of the Form II crystal of the meglumine salt of the compound of Formula 1 with the XRPD pattern after DVS analysis thereof.
31 is a graph showing an example of the XRPD pattern of the tromethamine salt of the compound of Formula 1;
32 shows a PLM picture of the tromethamine salt of the compound of Formula 1.
33 is a graph showing an example of an endothermic peak and a TGA curve appearing during DSC analysis of Form I crystals of tromethamine salt of the compound of Formula 1;
34 is a graph comparing the XRPD pattern of the tromethamine salt of Form I crystal after heating to 180° C. with the XRPD pattern of Form I crystal and Form II crystal.
35 is a graph showing an example of an endothermic peak and a TGA curve appearing during DSC analysis of Form II crystal of the tromethamine salt of the compound of Formula 1;
36 is a graph showing DVS results of Form II crystals of tromethamine salt of the compound of Formula 1;
37 is a graph comparing the XRPD pattern of Form II crystal of the tromethamine salt of the compound of Formula 1 with the XRPD pattern after DVS analysis thereof.
38 is a graph comparing the XRPD pattern of the potassium salt of the compound of Formula 1 with the XRPD pattern after storage according to each storage condition in the stability test according to Experimental Example 1. FIG.
39 is a graph comparing the XRPD pattern of the Form I crystal of the meglumine salt of the compound of Formula 1 and the XRPD pattern after storage according to each storage condition in the stability test according to Experimental Example 1. FIG.
40 is a graph comparing the XRPD pattern of the Form II crystal of the tromethamine salt of the compound of Formula 1 and the XRPD pattern after storage according to each storage condition in the stability test according to Experimental Example 1. FIG.
41 is a graph comparing the XRPD pattern of the free acid of the compound of Formula 1 with the XRPD pattern after storage according to each storage condition in the stability test according to Experimental Example 1. FIG.
FIG. 42 is a result of measuring the solubility of the free acid of the compound of Formula 1, the potassium salt of the compound of Formula 1, the Form I crystal of the meglumine salt, and the Form II crystal of the tromethamine salt in a buffer at pH 6.8.

Hereinafter, embodiments and examples of the present disclosure will be described in detail with reference to the accompanying drawings so that those of ordinary skill in the art may easily implement the present disclosure. However, the present application may be implemented in various forms and is not limited to the embodiments and examples described herein.

In the entire specification of the present application, when a certain part "includes" a certain constituent element, it means that other constituent elements may be further included rather than excluding other constituent elements unless otherwise specified.

The present invention provides a salt selected from the group consisting of potassium salt, sodium salt, L-arginine salt, L-lysine salt, ammonia salt, meglumine salt, and tromethamine salt as a salt of the compound of Formula 1 below. . Preferably, the salt is a potassium salt, meglumine salt, or tromethamine salt. Especially preferably, it is a potassium salt.

[Formula 1]

The compound of Formula 1 may be prepared according to the method disclosed in Korean Patent Nos. 1934651 and 1982667, and these documents are incorporated herein by reference in their entirety.

The salt of the compound of Formula 1 may be crystalline or amorphous, or may be prepared as a mixture thereof, but is preferably in a crystalline form.

In general, salts must have various physicochemical properties such as reproducibility of preparation of a specific crystal form, high crystallinity, stability of crystal form, chemical stability, and non-hygroscopicity in order to be applied to the pharmaceutical field.

From this point of view, the crystalline form of the salt of the compound of Formula 1 is excellent in stability and has physicochemical properties that are easy to formulate, and is particularly preferable in terms of hygroscopicity or solubility.

Preferably, the present invention provides a crystalline form of a potassium salt of the compound of Formula 1, a crystalline form of meglumine salt, or a crystalline form II of tromethamine salt. Particularly preferably, the compound of Formula 1 is provided in the form of a crystalline form of a potassium salt. The crystalline form of the potassium salt is X-ray powder diffraction containing characteristic peaks at 4 or more diffraction angles 2θ ± 0.2° selected from the group consisting of 4.6°, 9.0°, 9.3°, 16.0°, 19.8°, and 31.7° ( XRPD), characterized in that it exhibits a spectrum, it may be a Form I crystal. Preferably, the crystalline form exhibits an XRPD spectrum including characteristic peaks at diffraction angles 2θ ± 0.2° of 4.6°, 9.0°, 9.3°, 16.0°, 19.8°, and 31.7°, and more preferably, FIG. It has an XRPD spectrum as shown in 3.

The crystalline form of the potassium salt of the compound of Formula 1 may have an endothermic peak in the range of 80°C to 120°C, preferably 87°C to 113°C when analyzed by a differential scanning calorimeter (DSC) under a temperature rising condition of 10°C/min.

In one embodiment, the meglumine salt compound is X- containing characteristic peaks at 4 or more diffraction angles 2θ ± 0.2° selected from the group consisting of 10.2°, 16.6°, 17.0°, 25.0°, and 25.4°. It may be a Form I crystal showing a line powder diffraction (XRPD) spectrum. Preferably, the Form I crystal exhibits an XRPD spectrum including characteristic peaks at diffraction angles 2θ±0.2° of 10.2°, 16.6°, 17.0°, 25.0°, and 25.4°, and more preferably, FIG. 22 It has an XRPD spectrum as shown in.

The Form I crystal of the meglumine salt compound may have an endothermic peak in the range of 145° C. to 160° C., preferably 152° C. to 154° C. when analyzed by a differential scanning calorimeter (DSC) under a temperature rising condition of 10° C./min. .

In one embodiment, the tromethamine salt compound is characterized at 4 or more diffraction angles 2θ ± 0.2° selected from the group consisting of 3.9°, 7.9°, 13.7°, 16.3°, 17.3°, 17.7°, and 19.6°. It may be a Form II crystal showing an X-ray powder diffraction (XRPD) spectrum including peaks. Preferably, the Form II crystal exhibits an XRPD spectrum including characteristic peaks at diffraction angles 2θ ± 0.2° of 3.9°, 7.9°, 13.7°, 16.3°, 17.3°, 17.7°, and 19.6°, and further Preferably, it has an XRPD spectrum as shown in FIG. 31.

The Form II crystal of the tromethamine salt compound may have an endothermic peak in the range of 175°C to 190°C, preferably 182°C to 184°C when analyzed with a differential scanning calorimeter (DSC) under a temperature rising condition of 10°C/min. .

The present invention also provides a method for preparing the compound of formula 1 in salt form. Specifically, after dissolving a certain amount of the compound of Formula 1 in each solution, the same amount of base is added to the transparent solution. For example, the preparation method includes the steps of (a) adding a compound of Formula 1 in the form of a free acid to a solvent to obtain a solution; And (b) adding a base to the solution to prepare the compound of Formula 1 in the form of a salt. The solvent and base may be appropriately selected according to the salt type to be prepared. For example, the solvent includes, but is not limited to, methanol, ethanol, isopropanol, butanone, isobutanol, tetrahydrofuran, acetonitrile, methyl tert-butyl ether, acetone, water, ethyl acetate, or isopropyl acetate. Does not. The base includes, but is not limited to, NaOH, KOH, L-lysine, L-arginine, ammonia, meglumine, tromethamine, or nicotinamide.

In one embodiment, when preparing the compound of Formula 1 in the form of a potassium salt, ethyl acetate (EtOAc) may be used as the solvent, and potassium hydroxide may be used as the base.

The preparation method may further include the step of crystallizing the compound of Formula 1 in the salt form. The solvent used for the crystallization may be selected from the group consisting of ethyl acetate, acetone, acetonitrile, and a mixed solvent thereof. Alternatively, if the crystals do not precipitate, the solution may be concentrated or an anti-solution may be added.

In addition, the present invention provides a pharmaceutical composition comprising the salt or crystalline form, and a pharmaceutically acceptable additive.

In one embodiment, as disclosed in Korean Patent Nos. 1934651 and 1982667, the compound of Formula 1 exhibits an excellent inhibitory effect on TNF activity, and has been proven to be useful in the prevention or treatment of TNF overexpression diseases.

Accordingly, a pharmaceutical composition comprising a salt or a crystalline form of the compound of Formula 1 may be used for the prevention or treatment of a TNF overexpression disease by directly binding to the tumor necrosis factor (TNF) and inhibiting the activity of TNF.

For example, TNF overexpression diseases include rheumatoid arthritis, juvenile rheumatoid arthritis, inflammatory growth disease, Crohn's disease, ulcerative colitis, psoriasis, plaque psoriasis, juvenile plaque psoriasis, psoriatic arthritis, polyarticular pediatric idiopathic arthritis, Behcet's enteritis, ankylosing. Spondylitis, axial spondyloarthritis, pediatric bone-attached arthritis, osteoarthritis, multiple rheumatoid myalgia, multiple sclerosis, systemic lupus erythematosus, asthma, Sjogren's syndrome, pneumonia, chronic obstructive pulmonary disease, sarcoidosis, ring granulomatous, Wegener's granulomatosis, arteriosclerosis, Vasculitis, heart failure, myocardial infarction, kidney damage, nephritis, graft versus host disease, dementia, Alzheimer's disease, Parkinson's disease, pain, uveitis, Behcet's disease, purulent hypothyroidism, follicular erythema nasal inflammation, diabetic milk fat necrosis, necrotizing pyoderma , Sweet syndrome, subkeratinous pustular dermatosis, scleroderma, dermatomyositis, sepsis, and septic shock.

The present invention is to be described in more detail through the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[Preparation Example 1] Preparation of the compound of Formula 1

According to the method of Korean Patent No. 1934651 cited herein or a method similar thereto, a compound of Formula 1, that is, 2-(2,5-difluorophenyl)-3-carboxy-7-chloro-(4H) The free acid of -4-benzopyranone was prepared.

mp 147-150°C. ¹ H NMR (300 MHz, CDCl3): δ 7.26-7.43 (m, 3H), 7.69 (dd, J = 8.6, 1.8 Hz, 1H), 7.75 (d, J = 1.8 Hz, 1H), 8.41 (d, J = 8.6 Hz, 1H), 13.94 (s, 1H).

[Example 1]

96-well plate salt screening

An appropriate amount of 8 bases shown in Table 1 was dissolved in MeOH and diluted to 10 mL to prepare a 0.1 mol/L base solution.

Separately, a 0.1 mol/L solution was prepared by adding 362.0 mg of the compound of formula 1 in the form of a free acid to 12 mL of MeOH: DCM (5:1), and a 96-well plate having a volume of 100 μL in each well Was put into.

100 μL of the above base solution was added to the well and dried at room temperature. Then, 100 μL of the solvents of Table 1 were added to a 96-well plate, covered with parafilm having small pores, and evaporated under room temperature conditions.

menstruum base Methanol (MeOH) NaOH Ethanol (EtOH) KOH Isopropanol (IPA) L-lysine Butanone (MEK) L-arginine Isobutanol Ammonia Tetrahydrofuran (THF) Meglumine Acetonitrile (ACN) Tromethamine Methyl tert-butyl ether (MTBE) Nicotinamide Acetone (Acetone) - Water (H2O) - Ethyl acetate (EtOAc) - Isopropyl acetate (IPAC) -

After drying, the sample ^{was analyzed by 1} H NMR, XRPD, and PLM. The results are shown in Figs. 1 and 2, and Table 2 (however, in Figs. 1 and 2, only some of Table 2 are shown).

base
menstruum A B C D E F G H NaOH KOH L-Arg L-Lys ammonia Meglumine Nicotinamide Tromethamine One CR CR CR WCR CR CR CR CR MeOH 2 CR CR CR CR CR CR N/A CR EtOH 3 CR CR CR CR CR CR N/A CR IPA 4 CR CR CR CR CR CR N/A CR MEK 5 CR CR CR CR CR CR N/A CR Isobutanol 6 CR CR CR CR CR GL N/A CR THF 7 CR CR CR CR CR CR N/A CR ACN 8 CR CR CR CR CR CR N/A CR MTBE 9 CR CR CR CR CR WCR N/A CR Acetone 10 CR CR CR CR CR CR N/A CR H2O 11 CR CR CR CR CR WCR N/A CR EtOAc 12 CR CR CR CR CR CR N/A CR IPAC Note: CR = crystal; WCR = poor crystallinity; GL: glass state

White crystals were obtained in almost all wells except meglumine-THF (F6) (F6 is GL in Table 2; denoted as Glass state).

As shown in FIG. 1, when compared with the free acid form of the compound of Formula 1, chemical shifts in the compound prepared with NaOH, KOH, ammonia, L-arginine, L-lysine, meglumine and tromethamine are clearly chemical shifts (chemical shift) was found, which means that the bases formed a salt with the compound of formula 1.

As shown in FIG. 2, a characteristic peak of the free acid was found in the compound made of nicotinamide, indicating that the compound of Formula 1 did not form a salt with nicotinamide. In addition, from the XRPD of FIG. 2, it was found that a crystalline salt of NaOH salt, KOH salt, ammonia salt, L-arginine salt, L-lysine salt, meglumine salt and tromethamine salt having good crystallinity was obtained.

In the following, potassium salt, sodium salt, L-arginine salt, L-lysine salt, ammonia salt, meglumine salt and tromethamine salt were prepared based on the results of the 96-well plate salt screening.

After dissolving a certain amount of the compound of Formula 1 in each solution, the same amount of base was added to the transparent solution. If crystals did not precipitate, the solution was concentrated or an anti-solution was added.

[Example 2]

Preparation of potassium salt (K salt) of the compound of formula 1

According to the contents shown in Table 3, KOH (2 mol/L MeOH solution) as a base was added to an EtOAc solution of the compound of Formula 1 (free acid) to prepare a potassium salt of the compound of Formula 1. The obtained solid was dried at 60° C. and ^{analyzed by 1} HNMR, XRPD, PLM, DVS, TGA and DSC. The results are shown in Table 3 and FIGS. 3 to 7.

Lot # menstruum Volume (μL) Compound of formula 1
(mg) base
(Μl) result 33779-011A1 EtOAc 700 30.1 45 Form I crystal 33779-019G1 1000 44.7 68 Form I crystal

According to the XRPD pattern shown in FIG. 3, it was confirmed that the potassium salt of the compound of Formula 1 was obtained in a crystalline form having good crystallinity, and this crystal was defined as a Form I crystal (Table 3). The diffraction angle 2θ value representing a characteristic peak in the XRPD pattern is as follows.

4.6°, 9.0°, 9.3°, 16.0°, 19.8°, 31.7°

According to the PLM picture of the potassium salt of the compound of Formula 1 shown in FIG. 4, the potassium salt exhibited a small rod shape.

As a result of DSC analysis, as shown in Fig. 5, the DSC curve showed an endothermic peak with a starting point of about 87°C and a peak of about 113°C (melting with dehydration).

In addition, as a result of TGA analysis, as shown in FIG. 5, about 9% weight loss was confirmed at room temperature to about 120° C., which is due to the loss of water or residual solvent.

As a result of performing NMR, EtOAc/MeOH was not detected, and it was found that the potassium salt obtained therefrom was dihydrate (theoretical value 8.8%).

As a result of performing DVS, as shown in FIGS. 6 and 7, after drying at 40° C. under N2 flow, the I-form crystal changed to an amorphous anhydride, and absorbed 9.2% of moisture while forming a dihydrate at 30% RH ( Fig. 6). In addition, the I-form crystal of the potassium salt showed the same XRPD pattern even after the DVS test (FIG. 7). From this, it was confirmed that the potassium salt is stable at 10%RH or more.

[Example 3]

Preparation of sodium salt (Na salt) of the compound of formula 1

According to the contents shown in Table 4, sodium ethoxide or NaOH (2 mol/L MeOH solution) as a base was added to the EtOAc solution of the compound of Formula 1 to prepare a sodium salt of the compound of Formula 1. The obtained solid was dried at 60° C. and ^{analyzed by 1} HNMR, XRPD, PLM, TGA and DSC. The results are shown in Table 4 and FIGS. 8 to 11.

Lot # menstruum Volume (μL) Compound of formula 1
(mg) base result 33779-011B1 EtOAc 700 30.2 NaOH 450 μL Form I crystal 33779-019H1 1000 45.2 Sodium ethoxide 8.6 mg Form II crystal

According to the XRPD pattern shown in FIG. 8, it was confirmed that the sodium salt of the compound of Formula 1 was obtained in different crystalline forms depending on the reagent used in the preparation, and was defined as Form I crystal and Form II crystal, respectively (Table 4).

According to the PLM photograph of the sodium salt of the compound of Formula 1 shown in FIG. 9, both I-form and II-form crystals showed a needle shape.

As a result of TGA analysis, as shown in Figs. 10 and 11, weight loss of about 5% and about 4.4% was confirmed for Form I and Form II crystals at room temperature to about 130°C, respectively, which is due to the loss of residual solvent/water. will be.

In addition, as a result of DSC analysis, as shown in Figs. 10 and 11, it was confirmed that a wide endothermic peak appeared in both the I-form and II-form crystals according to the weight loss section of the TGA curve (melting with dehydration).

Since no solvent was detected in the NMR results, it was confirmed from the above results that both the I-form crystal and the II-form crystal are hydrates, and the moisture content of the sodium salt hydrate varies.

From the above experimental results, it was found that sodium salts, which may exist in various hydrate forms, are not suitable for future development.

[Example 4]

Preparation of L-arginine salt of the compound of formula 1

According to the contents shown in Table 5, L-arginine as a base was added to an EtOAc solution of the compound of Formula 1 to prepare an L-arginine salt of the compound of Formula 1. The obtained solid was dried at 60° C. and ^{analyzed by 1} HNMR, XRPD, PLM, TGA and DSC. The results are shown in Table 5 and FIGS. 12 to 14.

Lot # menstruum Volume (μL) Compound of formula 1
(mg) Base (mg) result 33779-011E1 EtOAc 700 30.1 15 Form I crystal

According to the XRPD pattern shown in FIG. 12, the L-arginine salt of the compound of Formula 1 was found to have poor crystallinity, and this was defined as Form I crystal (Table 5).

As a result of DSC analysis, as shown in Fig. 14, the DSC curve showed a broad endothermic peak with a starting point of 43°C and a peak point of 74°C (melting with dehydration).

In addition, as a result of TGA analysis, as shown in FIG. 14, about 6.8% weight loss was confirmed at room temperature to about 120° C., which is due to the loss of water or residual solvent.

As a result of performing NMR, EtOAc was not detected, and it was found that the L-arginine salt obtained therefrom was dihydrate (theoretical value was 6.7%).

From the above experimental results, it was found that the L-arginine salt has poor crystallinity, so it is not suitable for future development.

[Example 5]

Preparation of L-lysine salt of the compound of formula 1

According to the contents shown in Table 6, the L-lysine salt of the compound of Formula 1 was prepared.

Specifically, in Lot# 33779-031C1, 30.2 mg of the compound of Formula 1 was _{added to 1000 μL (0.1 mol/L) of an L-lysine MeOH/H 2} O solution, and then 2000 μL of MTBE was added to the solution. . The solution was stirred to obtain the L-lysine salt of the compound of formula 1 as a solid.

In Lot# 33779-031D1, 50 μL of an H2O solution of L-lysine was added to 700 μL of an acetone solution of the compound of formula (1). The solution was stirred to obtain the L-lysine salt of the compound of formula 1 as a solid.

The obtained solid was dried at 60° C. and ^{analyzed by 1} HNMR, XRPD, PLM, TGA and DSC. The results are shown in Table 6 and FIGS. 15 to 18.

Lot# Compound of formula 1 (mg) result 33779-031C1 30.2 Form I crystal 33779-031D1 45.2 Form II crystal

According to the XRPD pattern shown in FIG. 15, it was confirmed that the L-lysine salt of the compound of Formula 1 was obtained in different crystalline forms depending on the reagent used in preparation, and was defined as Form I crystal and Form II crystal, respectively (Table 6). . Both Form I and Form II crystals showed poor crystallinity.

In addition, according to the PLM photograph of the L-lysine salt of the compound of Formula 1 shown in FIG. 16, both I-form and II-form crystals showed irregular shapes.

As a result of TGA analysis, as shown in Figs. 17 and 18, weight loss of about 5.5% and about 3% was confirmed at room temperature to about 130°C, respectively, for Form I and Form II crystals, which is due to the loss of solvent/water. (Melting with dehydration).

In addition, as a result of DSC analysis, as shown in FIGS. 17 and 18, it was confirmed that both the I- and II-form crystals showed wide endothermic peaks according to the weight loss section of the TGA curve.

Since no solvent was detected in the NMR results, it was confirmed from the above results that both the I-form crystal and the II-form crystal are hydrates, and the water content of the L-lysine salt hydrate varies.

From the above experimental results, it was found that the L-lysine salt has poor crystallinity, so it is not suitable for future development.

[Example 6]

Preparation of ammonia salt of the compound of formula 1

According to the contents shown in Table 7, ammonia as a base was added to an EtOAc solution of the compound of Formula 1 to prepare an ammonia salt of the compound of Formula 1. The obtained solid was dried at 60° C. and ^{analyzed by 1} HNMR, XRPD, PLM, TGA and DSC. The results are shown in Table 7 and FIGS. 19 to 21.

Lot# menstruum Volume (μL) Compound of formula 1 (mg) Base (μL) result 33779-011C1 EtOAc 700 30.7 6.7 20% NH3 in water Form I crystal

According to the XRPD pattern shown in FIG. 19, the ammonia salt of the compound of Formula 1 was found to have poor crystallinity, and this was defined as Form I crystal (Table 7).

In addition, according to the PLM photograph of the ammonia salt of the compound of Formula 1 shown in FIG. 20, the ammonia salt of the compound of Formula 1 was in the form of agglomerated small crystals.

As a result of the DSC analysis, as shown in Fig. 21, three endothermic peaks (start point/peak point) of 87/109°C, 147/152°C, and 160/164°C were observed on the DSC curve.

As a result of TGA analysis, as shown in FIG. 21, a weight loss of about 4.9% at room temperature to 95°C and a weight loss of about 4.0% between 95°C and 145°C were observed, resulting in a total weight loss of 8.9% over two steps. Confirmed. This is due to the loss of solvent/water.

As a result of performing NMR, EtOAc was not detected, and it was found that the ammonia salt obtained therefrom was dihydrate (theoretical value was 9.2%).

From the above experimental results, it was found that the ammonia salt has poor crystallinity, and thus is not suitable for future development.

[Example 7]

Preparation of meglumine salt of the compound of formula 1

According to the contents shown in Table 8, meglumine as a base was added to a solution of EtOAc or Acetone of the compound of Formula 1 to prepare a meglumine salt of the compound of Formula 1. The obtained solid was dried at 60° C. and then ^{analyzed by 1} HNMR, XRPD, PLM, HSPLM, DVS, TGA and DSC. The results are shown in Table 8 and FIGS. 22 to 30.

Lot# menstruum Volume (μL) Compound of formula 1 (mg) base result 33779-011D1 EtOAc 700 30.8 16.5 mg Form II crystal 33779-019C1 Acetone 700 44.6 24.8 mg Form II crystal 33779-019E1 700 44.5 69 μL meglumine aqueous solution
(2 mol/L) Form I crystal 33779-042A1 700 45.2 69 μL meglumine aqueous solution
(2 mol/L) Form I crystal

According to the XRPD pattern shown in FIG. 22, it was confirmed that the meglumine salt of the compound of Formula 1 was obtained in different crystalline forms depending on the reagent used in preparation, and was defined as Form I crystal and Form II crystal, respectively (Table 8). . Both Form I and Form II crystals showed good crystallinity. The diffraction angle 2θ value representing the characteristic peak in the XRPD pattern of the I-form crystal is as follows.

10.2°, 16.6°, 17.0°, 25.0°, 25.4°

According to the PLM photograph of the meglumine salt of the compound of Formula 1 shown in FIG. 23, both I-form and II-form crystals showed irregular shapes.

As a result of DSC analysis of the Form I crystal of the meglumine salt, as shown in FIG. 24, the DSC curve showed an endothermic peak at about 152°C as a starting point and a peak at about 154°C (melting with decomposition). In addition, as a result of TGA analysis, as shown in FIG. 24, a weight loss of about 0.04% was confirmed at room temperature to about 150°C, indicating that there was no residual solvent. From the results of the thermal analysis, it can be seen that the Form I crystal is an anhydride.

As a result of performing DVS on the Form I crystal of meglumine salt, as shown in FIG. 27, 0.43% of water was absorbed at 80% RH. From this, it can be seen that the Form I crystal exhibits weak hygroscopicity. In addition, the I-form crystal showed the same XRPD pattern even after the DVS test (FIG. 28).

As a result of DSC analysis of the Form II crystal of meglumine salt, as shown in FIG. 25, the DSC curve showed two endothermic peaks (start point/peak point) of about 135/138°C and about 145/152°C. Particularly, as shown in the HSPLM of Fig. 26, the II-form crystal was melted at 135/138°C and converted to the I-form crystal. As a result of TGA analysis, as shown in FIG. 25, a weight loss of about 1.5% was confirmed at room temperature to about 120°C, indicating a loss of water or residual solvent. From the results of the thermal analysis, it can be seen that the Form II crystal is an anhydride.

As a result of performing DVS on the Form II crystal of meglumine salt, as shown in FIG. 29, 2.2% of moisture was absorbed at 80% RH. From this, it can be seen that Form II crystal exhibits hygroscopicity. In addition, the type II crystal showed the same XRPD pattern even after the DVS test (FIG. 30).

From the above experimental results, it was found that the Form II crystal of the meglumine salt is not suitable for future development due to its high hygroscopicity.

[Example 8]

Preparation of tromethamine salt of the compound of formula 1

According to the contents shown in Table 9, tromethamine as a base was added to a solution of EtOAc, acetone or ethanol of the compound of Formula 1 to prepare a tromethamine salt of the compound of Formula 1. The obtained solid was dried at 60° C. and ^{analyzed by 1} HNMR, XRPD, PLM, DVS, TGA and DSC. The results are shown in Table 9 and FIGS. 31 to 37.

Lot# menstruum Volume (μL) Of formula 1
Compound (mg) base result 33779-011F1 EtOAc 700 30.2 10.8 mg Form I crystal 33779-019A1 Acetone 700 44.7 250 μL MeOH/H2O solution
(0.5 mol/L) Form I crystal 33779-019B1 EtOH 3600 45.7 16.2 mg Form II crystal

According to the XRPD pattern shown in FIG. 31, it was confirmed that the tromethamine salt of the compound of Formula 1 was obtained in different crystalline forms depending on the reagent used in preparation, and was defined as Form I crystal and Form II crystal, respectively (Table 9). . Both Form I and Form II crystals showed good crystallinity. The diffraction angle 2θ value representing a characteristic peak in the XRPD pattern of the II type crystal is as follows.

3.9°, 7.9°, 13.7°, 16.3°, 17.3°, 17.7°, 19.6°

According to the PLM photograph of the tromethamine salt of the compound of Formula 1 shown in FIG. 32, the I-form crystal was in a plate shape, and the II-form crystal was in a needle shape.

No solvent was detected in the NMR results.

As a result of TGA analysis of the Form I crystal of the tromethamine salt, as shown in FIG. 33, a weight loss of about 1.4% was confirmed at room temperature to about 170° C., indicating a loss of water. DSC analysis showed two endothermic peaks (start point/peak point) of 145/160° C. and 185/186° C., indicating loss of water and melting of Form II crystals, respectively. As shown in Fig. 34, after heating the I-form crystal to 180°C, a II-form crystal was obtained. In addition, from the above analysis results, it can be seen that Form I crystal is a hydrate.

As a result of TGA analysis of the Form II crystal of the tromethamine salt, as shown in FIG. 35, a weight loss of about 0.6% was confirmed at room temperature to about 150°C, indicating a loss of water or residual solvent. As a result of DSC analysis, one endothermic peak at a starting point of 182°C and a peak of 184°C was shown (melting with decomposition).

As a result of performing DVS on the Form II crystal, as shown in FIG. 36, the Form II crystal absorbed 0.55% of water at 80% RH. From this, it can be seen that Form II crystal exhibits weak hygroscopicity. In addition, the type II crystal showed the same XRPD pattern even after the DVS test (FIG. 37).

From the above experimental results, it was found that the I-form crystal of the tromethamine salt can be transformed into a II-form crystal, and thus its stability is poor, and thus it is not suitable for future development.

From the experiments of Examples 1 to 8, it was confirmed that the compound of Formula 1 forms a salt with KOH, NaOH, L-arginine, L-lysine, ammonia, meglumine, and tromethamine. Potassium salt showed good crystal form as a dihydrate form, and meglumine salt and tromethamine salt showed good crystallinity and high melting point (>150° C.) as anhydride forms. However, it was found that the Form II crystal of the meglumine salt showed high hygroscopicity, and the Form I crystal of the tromethamine salt could be transformed into a Form II crystal, so that it was not suitable for future development. The sodium salt, L-arginine salt, L-lysine salt, and ammonia salt were in hydrate form and showed poor crystallinity. Accordingly, the KOH salt, the meglumine salt of the Form I crystal, and the tromethamine salt of the Form II crystal were selected, and solubility and solid stability tests were performed.

[Experimental Example 1] Stability test

Stability tests were performed on the compound of formula 1 (free acid form), potassium salt, meglumine salt of Form I crystal, and tromethamine salt of Form II crystal. The results are shown in Table 10 and FIGS. 38 to 41. As a result of XRPD and HPLC analysis, the samples were physicochemically stable for 7 days at 40°C/75%RH and 60°C.

sample Purity (Area %) Early 40℃ /75% RH-7day 60℃-7days Compound of formula 1 (free acid) 100 100 99.93 Potassium salt 99.97 99.97 99.90 Meglumine salt 99.94 99.93 99.90 Tromethamine salt 99.97 99.95 99.98

[Experimental Example 2] Solubility Measurement

Solubility tests were performed for the compound of formula 1 (free acid form), potassium salt, meglumine salt of Form I crystal, and tromethamine salt of Form II crystal. The results are shown in FIG. 42.

Specifically, a compound of Formula 1, a potassium salt, a meglumine salt of a type I crystal, and a tromethamine salt of a type II crystal were added to various buffers and observed. All of these samples had very low solubility in pH 1.2 buffer (<0.5 mg/mL). The solubility of the three salts was similar to the solubility of the compound of Formula 1 (free acid), and in the pH 6.8 buffer, the solubility of the meglumine salt and the tromethamine salt did not improve compared to the free acid form, but the potassium salt was solubility. It was confirmed that was improved (FIG. 42).

From the results of Experimental Examples 1 and 2, it was confirmed that the potassium salt is most suitable for future development.

[Reference Example] Equipment and measurement conditions used

(1) X-ray powder diffraction test (XRPD)

Solid samples were analyzed using a powder X-ray diffractometer (Bruker D8 advance) equipped with a LynxEye detector. The instrument parameters used are as follows.

Scan: 3°(2θ) to 40°(2θ)

Amplification (interval): 0.02°(2θ)

Scan speed: 0.3 sec/step

Voltage: 40 KV

Current: 40 mA

Rotation: On

Sample holder: Zero-background sample holder

(2) Polarization microscopy analysis (PLM)

PLM analysis was performed using a polarization microscope ECLIPSE LV100POL (Nikon, JPN).

(3) Thermogravimetric Analysis (TGA)

TGA was performed with Discovery TGA 55 (TA Instruments, US). The sample was placed in an open tarred aluminum pan and weighed automatically, and the sample was inserted into a TGA heater. The sample was heated to the final temperature at a rate of 10° C./min.

(4) Differential Scanning Calorimetry (DSC)

DSC analysis was performed with a Discovery DSC 250 (TA Instruments, US). The sample was placed on a DSC pinhole pan, the weight was accurately recorded, and then heated to the final temperature at a rate of 10°C/min.

(5) ^One H NMR analysis

¹ H NMR was performed using a Bruker Advance 300 equipped with an automated sampler (B-ACS 120).

(6) Dynamic moisture adsorption analysis (DVS)

DVS was determined using DVS Intrinsic (SMS, UK). About 20-50 mg of the compound was transferred to the SMS dynamic moisture adsorption model DVS and the weight change with varying atmospheric humidity at 25°C was recorded. The instrument parameters used are as follows.

Equilibrium time: 60 min

RH (%) measuring point:

Adsorption: 0, 10, 20, 30, 40, 50, 60, 70, 80, 90

Desorption: 90, 80, 70, 60, 50, 40, 30, 20, 10, 0

(7) Hot stage polarized light microscope (HSPLM)

HSPLM analysis was performed on a hot stage (Likam, LTS420) equipped with a polarized light microscope (Nikon, ECLIPSE LV100POL).

(8) High performance liquid chromatography (HPLC)

HPLC was performed using an Agilent 1260 for solubility testing. The instrument parameters used are as follows.

Column: Eclipse plus C18, 3.5 μm, 4.6*100 mm

Column temperature: 30℃

Mobile phase A: 0.1% TFA in H2O; B: 0.1% TFA in ACN

Gradient condition (% of B): 35%(0 min), 100%(8 min), 100%(10 min)

Flow: 1.0 mL/min

Injection volume: 5 μL

UV wavelength: 240 nm (Ref: off)

Post time: 3 min

Claims (7)

The compound of formula 1 in the form of a salt selected from the group consisting of potassium salt, sodium salt, L-arginine salt, L-lysine salt, ammonia salt, meglumine salt, and tromethamine salt.
[Formula 1]

A compound of formula 1 in the form of a potassium salt, meglumine salt, or tromethamine salt.
[Formula 1]

Potassium salt compound of the following formula 1.
[Formula 1]

The X-ray powder diffraction according to claim 3, comprising characteristic peaks at 4 or more diffraction angles 2θ±0.2° selected from the group consisting of 4.6°, 9.0°, 9.3°, 16.0°, 19.8°, and 31.7°. (XRPD) A crystalline form, characterized by showing a spectrum, the compound.
The compound according to claim 4, characterized in that it has an endothermic peak in the range of 80°C to 120°C when analyzed by a differential scanning calorimeter (DSC) under a temperature rising condition of 10°C/min.
(a) obtaining a solution by adding a compound of Formula 1 to ethyl acetate (EtOAc); And
(b) preparing a compound of Formula 1 in the form of a potassium salt by adding potassium hydroxide to the solution;
A method for preparing a potassium salt compound of Formula 1 according to any one of claims 3 to 5, including.
Comprising the compound according to any one of claims 1 to 5 as an active ingredient, rheumatoid arthritis, juvenile rheumatoid arthritis, inflammatory growth disease, Crohn's disease, ulcerative colitis, psoriasis, plaque psoriasis, juvenile plaque psoriasis, psoriasis Arthritis, polyarticular pediatric idiopathic arthritis, Behcet's enteritis, ankylosing spondylitis, axial spondyloarthritis, pediatric bone-attached arthritis, osteoarthritis, multiple rheumatoid myalgia, multiple sclerosis, systemic lupus erythematosus, asthma, Sjogren syndrome, pneumonia, chronic obstructive pulmonary disease , Sarcoidosis, ring granulomatosis, Wegener's granulomatosis, arteriosclerosis, vasculitis, heart failure, myocardial infarction, kidney damage, nephritis, graft-versus-host disease, dementia, Alzheimer's disease, Parkinson's disease, pain, uveitis, Behcet's disease, purulent thyroiditis, pores A pharmaceutical composition for the prevention or treatment of a TNF overexpression disease selected from the group consisting of scarlet nasal inflammation, diabetic milk fat necrosis, gangrene pyoderma, sweet syndrome, subkeratinous pustular dermatosis, scleroderma, dermatomyositis, sepsis and septic shock.

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