COMPOUND, PROCESS FOR PREPARING THE COMPOUND, AND PHARMACEUTICAL COMPOSITION

Abstract

COMPOUND, PROCESS FOR PREPARING THE COMPOUND, AND PHARMACEUTICAL COMPOSITION. The present invention relates to 1,5-hydrate of 1-{(2S)-2-amino-4-[2,4-bis(trifluoromethyl)-5,8-dihydropyrido[3,4-d]pyrimidin-7(6H)-yl]-4-oxobutyl}-5,5-difluoropiperidin-2-one tartrate, a process for preparing the same, and a pharmaceutical composition for inhibiting DPP-IV, which comprises said compound as the active component.

Description

1 / 21 “COMPOUND, PROCESS FOR PREPARING THE COMPOUND, AND PHARMACEUTICAL COMPOSITION” Technical field

[001] The present invention relates to 1,5-hydrate of 1{(2S)-2-amino-4-[2,4-bis(trifluoromethyl)-5,8-dihydropyrido[3,4-d]pyrimidin7(6H)-yl]-4-oxo-butyl}-5,5-difluoropiperidin-2-one tartrate represented by the following formula 1 (hereinafter referred to as “Compound 1”), and a process for preparing the same. nh2(1) Fundamentals of the Technique

[002] Compound 1 is described in Korean Patent Application No. 10-2006-0029138, which is incorporated herein by reference in its entirety, and exhibits superior inhibitory activity against dipeptidyl peptidase-IV (DPP IV) and is thus useful as an agent for treating diabetes. Typical examples of diseases caused by DPP IV may include, but are not limited to, diabetes, obesity, etc. Among diabetes, it is particularly useful for the treatment and prevention of type II diabetes. The term “treatment” as used above means that when the compound is used for individuals who manifest symptoms of a disease, it can stop or delay the progression of the disease; and the term “prevention” as used above means that when the compound is used for individuals who do not manifest symptoms of a disease but are at risk of developing the disease, it can stop or delay the onset of the disease.

[003] Investigating the physical and chemical properties of a new drug is necessary for efficient and successful development. Petition 870210062456, dated 09 / 07 / 2021, page 12 / 43 2 / 21 of a new drug. In particular, through the study of the presence of polymorphic and pseudopolymorphic forms of the drug and differences in physical and chemical properties between respective polymorphic forms, the preferred crystal form of the drug can be selected in view of the pharmaceutical aspect (Remington's Pharmaceutics, Chapter 75 Preformulation); (Byrn, SR, Solid State Chemistry of Drugs, Academic Press, New York, 1982). When the polymorphic forms are present in solution, they are chemically identical, but in the solid state they respectively have definitively different x-ray diffraction patterns and exhibit differences in various physical and chemical properties. In particular, respective polymorphic forms may have differences in bioavailability due to differences in dissolution rates, and exhibit unexpected properties in terms of thermodynamic stability.

[004] When a certain drug is present in the form of polymorphic forms, crystal forms having different structures can be obtained depending on the recrystallization conditions such as recrystallization solvent, drug concentration, heating and cooling rates, temperature, stirring rate, and the like, during the procedures to prepare the drug. Therefore, in order to obtain the same crystal form, special attention is required to the management of manufacturing procedures.

[005] Hydrates, as one of the pseudopolymorphic forms, comprise a water molecule within the drug crystal and possess a different crystal form than an anhydrate. Differences in crystal structure can be distinguished by X-ray diffraction pattern. As in hydrates only physical properties such as crystallinity, hygroscopic properties, melting point, solubility, dissolution rate, etc., are altered without any change in chemical properties providing pharmacological effects, they have a very important significance in the pharmaceutical aspect, as polymorphic forms (Morris, KR et al., Int. J. Pharm., 108, 1994, 15-206). Petition 870210062456, dated 09 / 07 / 2021, page 13 / 43 3 / 21

[006] The understanding that is understood to date from various references relating to the technical field to which the present invention belongs is that there is no general tendency, for example, to prefer the hydrate to the anhydrate or vice versa, for the improvement of pharmaceutical properties including drug stability, hygroscopic properties, etc. Finally, the determination of the forms having the optimum pharmaceutical properties for respective compounds should be made by an expert in the relevant technical field through continuous case-by-case study.

[007] In particular, it can never be anticipated which of the contemplable forms of a certain drug, i.e., free compound, salt, anhydrate, and hydrate, might exhibit stability with hygroscopic properties that are not altered depending on the surrounding moisture. Additionally, among the hydrates, the most stable hydration number cannot be predicted. And even if the hydration number is the same, it is also unpredictable which crystal form might be the most stable. This is a phenomenon that is inconsistently revealed since it cannot be anticipated and belongs to the experimental area that can only be confirmed through coated experiments. Description of the Invention Technical Problem

[008] Then, the present inventors conducted intensive study to provide a stable polymorphic form or pseudopolymorphic form of Compound 1. As a result, surprisingly we discovered that the 1,5 tartrate salt hydrate of Compound 1 exhibits superior stability against changes in relative humidity compared to the anhydrate or other hydrates having a similar hydration number, and thus completed the present invention. Until now, the crystal form of Compound 1 has never been publicly described. Solution to the Problem

[009] Therefore, the present invention provides 1,5 hydrate of Petition 870210062456, dated 09 / 07 / 2021, p. 14 / 43 4 / 21 tartrate salt of Compound 1.

[010] In addition, the present invention provides a process for preparing 1,5 tartrate salt hydrate of Compound 1. Advantageous Effects of the Invention

[011] As the 1,5 tartrate salt hydrate of Compound 1 obtained as a crystal form developed according to the present invention is superior to similar crystal forms in terms of stability, particularly storage stability, it can be very advantageously used in the preparation of pharmaceutical compositions containing Compound 1 as an active component. That is, the 1,5 hydrate according to the present invention neither loses any water molecules in the crystal nor accepts more water molecules across a wide range of relative humidity to maintain its water content, and therefore does not show a substantial change in weight depending on changes in humidity. In the case of unstable crystal forms, the water content thereof may vary with environments or additives during the formulation and storage process.For example, in the quantification of the standard material and the sample for quantification purposes, if the experiment is not conducted in a drying environment, some experimental error may occur, leading to quality control problems. However, since the water content of the 1,5-hydrate according to the present invention does not vary significantly with environments in a wide range of relative humidity, a product with a uniform standard can always be obtained during the formulation and storage process, and additionally, the error in quality control is very small. As above, the 1,5-hydrate according to the present invention shows a great advantage in terms of handling and quality control.

[012] Additionally, 1,5 tartrate salt hydrate of Compound 1 according to the present invention does not show any change in crystal form depending on the change in humidity. On the contrary, the present inventors have identified that 0,5 hydrate and anhydrate of Petition 870210062456, dated 09 / 07 / 2021, page 15 / 43 5 / 21 tartrate salt of Compound 1 absorbs a lot of water when the relative humidity is high, to be converted into a more stable 1,5 hydrate according to the present invention. It was also identified that even in the experiment for acceleration stability (40°C / 75% RH) the 0,5 hydrate is converted into 1,5 hydrate after a few hours.

[013] In addition, it is possible to control the preparation of 1,5 tartrate salt hydrate of Compound 1 according to various methods as provided in the present invention. Brief Description of the Drawings

[014] Figure 1 represents the x-ray powder diffraction (XRD) spectrum of crystal form I as the 1,5 tartrate salt hydrate of Compound 1.

[015] Figure 2 represents the infrared spectroscopy (FT-IR) spectrum of crystal form I as the 1,5 tartrate salt hydrate of Compound 1.

[016] Figure 3 represents the result of Differential Scanning Calorimetry (DSC) or Thermogravimetric (TG) analyses of crystal form I as the 1,5 tartrate salt hydrate of Compound 1.

[017] Figure 4 represents the dynamic vapor adsorption / desorption isotherm of crystal form I as the 1,5 tartrate salt hydrate of Compound 1.

[018] Figure 5 represents the X-ray powder diffraction (XRD) spectrum of crystal form II as the 0.5 tartrate salt hydrate of Compound 1.

[019] Figure 6 represents the infrared spectroscopy (FT-IR) spectrum of crystal form II as the 0.5 tartrate salt hydrate of Compound 1.

[020] Figure 7 represents the result of Differential Scanning Calorimetry (DSC) or Thermogravimetric (TG) analyses of crystal form II as the 0.5 tartrate salt hydrate of Compound 1. Petition 870210062456, dated 09 / 07 / 2021, page 16 / 43 6 / 21

[021] Figure 8 represents the dynamic vapor adsorption / desorption isotherm of crystal form II as the 0.5 tartrate salt hydrate of Compound 1.

[022] Figure 9 represents the X-ray powder diffraction (XRD) spectrum of crystal form II as the 0.5 tartrate salt hydrate of Compound 1, during the course of the moisture adsorption study.

[023] Figure 10 represents the x-ray powder diffraction (XRD) spectrum of crystal form III as the tartrate anhydrate salt of Compound 1.

[024] Figure 11 represents the infrared spectroscopy (FT-IR) spectrum of crystal form III as the tartrate anhydrate salt of Compound 1.

[025] Figure 12 represents the result of Differential Scanning Calorimetry (DSC) or Thermogravimetric (TG) analyses of crystal form III as the tartrate salt anhydrate of Compound 1.

[026] Figure 13 represents the dynamic vapor adsorption / desorption isotherm of crystal form III as the tartrate salt anhydrate of Compound 1.

[027] Figure 14 represents the moisture adsorption curve of crystal form III as the tartrate salt anhydrate of Compound 1, with time passing at normal temperature and normal humidity.

[028] Figure 15 represents the X-ray powder diffraction (XRD) spectrum of crystal form III as the tartrate anhydrate salt of Compound 1, during the moisture adsorption study.

[029] Figure 16 represents the X-ray powder diffraction (XRD) spectrum obtained after conducting the Test Example 8 experiment, which indicates the XRD of crystal form II, the XRD of the crystal form II sample after storage for 8 weeks at 60°C / 5% RH, the XRD of the crystal form II sample after 8 weeks of storage at 40°C / 75% Petition 870210062456, dated 09 / 07 / 2021, page 17 / 43 7 / 21 of UR, and the XRD of crystal form I, in order from the bottom. Method for the Invention

[030] As indicated to respectively possess the characteristic crystal forms according to the present invention, in the present application the 1,5 tartrate salt hydrate of Compound 1 is designated as “crystal form I”; the 0,5 hydrate is designated as “crystal form II”; and the anhydrate is designated as “crystal form III”. Herein, the water content of 1,5 hydrate is 3.0~5.5%, that of 0,5 hydrate is 1.0~2.5%, and that of anhydrate is 0~1.0%.

[031] The crystallinities of crystal forms I, II and III are different from each other as can be identified from x-ray diffraction diagrams shown in Figures 1, 5 and 10.

[032] As a result of Differential Scanning Calorimetry analysis for crystal form I, the endothermic peak that occurs with the release of water contained in the crystal lattice at 90~130°C has a large section in which the melting point is included, and the endothermic peak occurs again in the 130~160°C section with the passage through the chemical dehydration of the tartrate salt of Compound 1. In addition, thermogravimetric analysis shows a weight loss of approximately 4.0%, which is equivalent to the water contained in the lattice in the first endothermic section of Differential Scanning Calorimetry, and a weight loss of approximately 2.5%, which is equivalent to the water removed by dehydration from the tartrate salt structure of Compound 1 in the second endothermic section (Figure 3). Analysis of crystal forms II and III by Differential Scanning Calorimetry shows three endothermic points.Specifically, the first endothermic point occurs with the release of water contained in the lattice, the second endothermic point occurs with the melting point, and the third endothermic point occurs through a chemical dehydration reaction. Additionally, according to thermogravimetric analysis, the first endothermic section shows a weight loss equivalent to the water contained in the lattice (approximately 1.3% in the form of crystal II, e. Petition 870210062456, dated 09 / 07 / 2021, page 18 / 43 8 / 21 approximately 0.6% in crystal form III), and the third endothermic section shows a weight loss of approximately 2.4%, which is equivalent to the water removed by dehydration (Figures 7 and 12). As the water content in the tartrate salt network of Compound 1 as above was consistent with the water content quantified by the Karl-Fischer method (Mettler Toledo DL37 KF Coulometer), it was proven that the endothermic peaks were caused by the evaporation of water molecules.

[033] Hereinafter, we intend to explain that crystal form I according to the present invention has unexpected superior characteristics from any aspect compared with other crystal forms II or III, more specifically by comparing the characteristics of the respective crystal forms. Crystal form I

[034] The present invention relates to the 1,5 tartrate salt hydrate crystal form of Compound 1. As a result of the analysis of its properties, crystal form I exhibits the following characteristics: (a) The water content of crystal form I is in the range of 3.0~5.5%. (b) The characteristic peak values ​​(2θ) of the XRD spectrum measured in CuKa, 40kV, 30mA are 15, 18, 20, 21 and 23°. (c) The infrared (IR) spectrum shows characteristic absorbance at approximately 3591, 3401, 3128, 1712, 1655, 1636, 1229, 1205, 1129 and 1058 cm-1. (d) The Differential Scanning Calorimetry spectrum shows endothermic peaks in two wide temperature ranges of approximately 90-130°C and 130-160°C. (e) The water content measured by the Karl-Fisher method is approximately 4.0%. (f) When the temperature is raised from 25°C to 250°C, the Petition 870210062456, dated 09 / 07 / 2021, page 19 / 43 9 / 21 weight loss of approximately 4.0% and approximately 2.5% occurs in the ranges of 70~110°C and 140~170°C, respectively (This is the result obtained from TG in Figure 3. As the energy change occurs soon after the weight change, there is generally a difference between the DSC and TG results.). (g) The tartrate salt of Compound 1 can be crystallized from water, acetonitrile / water, ethanol / water, ethanol / hexane or ethyl acetate / hexane solvents, and preferably crystallized from water. (h) Crystal form I can be prepared by subjecting crystal form II or III to moisture absorption. (i) Weight change is absent or, if present, 0.8% or less depending on the change in external humidity in the range of 5~95% RH, and the crystal form is not altered by the change in humidity. Crystal form II

[035] In order to find other crystal forms different from crystal form I, the present inventors prepared 0.5 hydrate tartrate salt of Compound 1 (crystal form II). Although the 0.5 hydrate is the same tartrate salt of the same compound and has a similar hydration number, it shows unstable storage stability as follows, compared to the 1.5 hydrate (crystal form I). (a) The water content of crystal form II is in the range of 1.0~2.5%. (b) The characteristic peak values ​​(2θ) of the XRD spectrum measured in CuKa, 40kV, 30mA are 14, 15, 17, 18, 19, 21 and 23°. (c) Infrared (IR) spectrum shows characteristic absorbance at approximately 3455, 2891, 1721, 1655, 1571, 1228, 1209, 1131, 1086 and 1059 cm-1. (d) The Differential Scanning Calorimetry spectrum shows endothermic peaks in three wide temperature ranges of about 80~115°C, 115~135°C and 135~173°C, and the melting point at about 117°C. Petition 870210062456, dated 09 / 07 / 2021, p. 20 / 43 10 / 21 (e) The water content measured by the Karl-Fisher method is approximately 2.0%. (f) When the temperature is raised from 25°C to 250°C, weight loss of approximately 1.3% and approximately 2.4% occurs in the range of 70-104°C and 137-168°C, respectively. (g) A weight change of 4.0% or more occurs upon a change in external humidity in the range of 5-95% RH, and moisture is rapidly absorbed from 45% RH, resulting in a weight increase of 3.7% at 75% RH. If a total humidity of 4.0% or more is present, crystal form II is converted to crystal form I. It has been shown that crystal form II is converted to crystal form I from about 60% RH (see Figures 8 and 9). That is, the crystal form varies with the change in humidity to reach the more stable crystal form I. In addition, it is converted to crystal form I within 8 weeks in the acceleration stability test (40°C / 75% RH). Crystal form III

[036] In order to find other crystal forms different from crystal form I, the present inventors prepared tartrate salt anhydrate of Compound 1 (crystal form III). The anhydrate shows unstable storage stability as follows, compared to the 1,5 hydrate (crystal form I). (a) The water content of crystal form III is in the range of 0~1.0%. (b) The characteristic peak values ​​(2θ) of the XRD spectrum measured in CuKa, 40kV, 30mA are 6, 17, 21, 23, 24, 26 and 30°. (c) The infrared (IR) spectrum shows characteristic absorbance at approximately 3470, 3187, 2940, 1640, 1570, 1229, 1206, 1130 and 1056 cm-1. (d) Differential Scanning Calorimetry Spectrum shows Petition 870210062456, dated 09 / 07 / 2021, page 21 / 43 11 / 21 the endothermic peaks in three broad temperature ranges of approximately 65~100°C, 100~130°C and 132~170°C, and the melting point at approximately 104°C. (e) The water content measured by the Karl-Fisher method is approximately 0.1%. (f) When the temperature is raised from 25°C to 250°C, weight loss of approximately 0.6% and approximately 2.4% occurs in the range of 62~110°C and 120~173°C, respectively. (g) A weight change of 3.5% or more occurs upon a change in external humidity in the range of 5~95% RH, and moisture is rapidly absorbed from 5% RH, so that crystal form III is converted to crystal form I from about 15% RH (see Figures 13, 14 and 15). That is, the crystal form varies with the change in humidity to arrive at the more stable crystal form I.

[037] The above results suggest that in the relative humidity range under which the formulation is conventionally practiced, crystal forms II and III are unstable and thus automatically converted into crystal form I. This tendency is shown most prominently in the case of crystal form III.

[038] The 1,5 tartrate salt hydrate of Compound 1 according to the present invention exhibits strong DPPIV inhibitory activity in the same manner as the corresponding free base described in Korean Patent Application No. 10-2006-0029138. In addition, the 1,5 hydrate of the present invention shows improved physical and chemical properties when compared to crystal forms having other hydration states. Therefore, the 1,5 hydrate according to the present invention is considerably easier to handle, quality control, and formulate compared to crystal forms having other hydration states.

[039] The 1,5 hydrate according to the present invention has the DPP-IV inhibitory activity as mentioned above, and therefore, Petition 870210062456, dated 09 / 07 / 2021, page 22 / 43 12 / 21 can be formulated for convenient administration in the veterinary and pharmaceutical fields. The formulation can be conducted according to techniques and methods known in the art in relation to other formulations having DPP-IV inhibitory activity, particularly with reference to the description in Korean Patent Application No. 10-2006-0029138, which is incorporated herein by reference in its entirety.

[040] Therefore, a pharmaceutical composition for inhibiting DPP-IV, comprising the 1,5 hydrate according to the present invention as the active component together with a pharmaceutically acceptable carrier, is covered by the scope of the present invention, the composition according to the present invention is characterized by the fact that it is used particularly for the treatment and prevention of diabetes or obesity.

[041] The present invention is illustrated in more detail by means of the following Examples and Test Examples. However, the following Examples and Test Examples are provided only to aid understanding of the present invention but are not intended to limit the scope of the present invention in any way by these Examples and Test Examples. Example 1 Preparation of 1.5 hydrate tartrate salt of Compound 1 (crystal form I)

[042] 1.87 kg of compound 2 was dissolved in approximately 9 L of ethanol. 0.94 kg of SOC12 was added at 0-10°C and then stirred while maintaining a low temperature. After concentration under pressure Petition 870210062456, dated 09 / 07 / 2021, p. 23 / 43 13 / 21 reduced, the concentrate was dissolved in 11.2 L of MTBE (methyl tert-butyl ether), and the resulting mixture was adjusted with 10 N NaOH solution to pH 7-8. After separation of the layers, the aqueous layer was extracted with approximately 3.7 L of MTBE and twice with 3.7 L of MTBE, and then concentrated under reduced pressure. The resulting turbid brown solution was dissolved in 12 L of ethanol, 0.47 kg of L-tartaric acid dissolved in approximately 1.5 L of water was added, and then stirred for 1 hour. The resulting crystalline fluid paste was filtered, washed with water and ethanol (1:8), and then dried to obtain 1.13 kg (97.5% yield) of the title compound. 1H NMR (500 MHz, CD3OD) δ 2.38 (m, 2H), 2.59 (m, 2H), 2.82 - 2.99 (m, 2H), 3.11 (bt, 1H), 3.21 (bt, 1H), 3.50 - 3.55 (m, 1H), 3.72 - 3.91 (m, 5H), 3.98 (t, J=5.2 Hz, 1H), 4.38 (s, 2H), 4.97 - 5.00 (m, 2H). Example 2 Recrystallization of 1,5 tartrate salt hydrate of Compound 1 (crystal form I) from water

[043] 50 g of tartrate salt of Compound 1 obtained from Example 1 was added to 250-500 ml of water and dissolved in water while adjusting the solution with 10 N NaOH to pH 6-7. 11.7 g of L-tartaric acid dissolved in 23.5 ml of water was stirred, and crystals were obtained by varying the temperature, stirring rate, and stirring time as shown in the following Table 1. Then, the crystals were filtered and dried to obtain crystal form I. The stirring rate was varied in the range of 50-400 rpm, and the temperature was varied in the range of 5-32°C. The volume of water used for recrystallization, the stirring rate, the temperature, and the stirring time are shown in the following Table 1. [Table 1] Input Volume (mL) RPM Temperature Time (h) Yield (%) 1 250 50 25 0.5 96.0 (48.0g) 2 250 100 25 1 92.8 (46.4g) 3 250 400 25 5 93.6 (46.8g) 4 375 400 25 1 92.4 (46.2g) Petition 870210062456, dated 09 / 07 / 2021, page 24 / 43 14 / 21 5 500 50 25 5 46.6 (23.3g) 6 500 400 25 3 88.2 (44.1g) 7 375 225 31.8 2.75 94.0 (47.0g) 8 375 225 15 2.75 91.0 (45.5g) 9 250 50 5 0.5 81.4 (40.7g) 10 250 400 5 5 101.8 (50.9g) Example 3 Recrystallization of 1,5 tartrate salt hydrate of Compound 1 (crystal form I) from the mixed solvent.

[044] 5 g of tartrate salt of Compound 1 was dissolved in 25-60 ml of solvent mixtures comprising water and acetonitrile in different ratios. Crystals were precipitated by varying the temperature in the presence or absence of agitation, filtered and dried to recrystallize the tartrate salt of Compound 1. The conditions used for recrystallization are listed in the following Table 2. [Table 2] Acetonitrile / water input Agitation Temperature 1 4 / 1 No 25 3 6 / 1 No 25 4 6 / 1 No 0 5 8 / 1 No 25 6 8 / 1 Yes 25 8 1 / 1 No 25 9 1 / 1 Yes 20 Example Test 1 Powder X-ray diffractometry

[045] Approximately 20 mg of the sample was loaded into the sample holder and mounted on the powder X-ray diffractometer to obtain the diffraction pattern in the range of 3~40° / 2θ. The diffraction patterns as obtained are attached to this specification as Figures 1, 5 and 10, respectively. Specific conditions for analysis are as follows. Instrument: Bruker 4D Endeavor Time per step: 0.3 s Step size: 0.03° Petition 870210062456, dated 09 / 07 / 2021, page 25 / 43 15 / 21 Scanning mode: step Voltage / Current: 40 kV / 30 mA Target Cu (Ni Filter) Divergence gap: 0.3 Detector: PSD: LynxEye Instrument: Philips X-ray generator (PW1710) Time per step: 0.5 s Step size: 0.03° Scanning mode: step Voltage / Current: 40 kV / 30 mA Target Cu (Ni Filter) Slot size: 1.0 mm Detector Slots: 0.15 mm, 1.0 mm Example Test 2 Infrared spectroscopy

[046] Infrared spectra for respective crystal forms according to the present invention were obtained using a ZASCO FT-IR 4200 equipped with a DTGS detector. The resolution of the respective spectra was 4 cm⁻¹, and the scan number was 16. In the present Test Example, 1-2 mg of the sample was positioned in the ATR (Attenuated Total Reflectance) accessory, and the equipment was operated to obtain the spectrum. Supporting data were obtained by operating the equipment without any material in the ATR. The spectra thus obtained are attached to the present specification as Figures 2, 6, and 11, respectively. Example Test 3 Differential Scanning Calorimetry (DSC)

[047] Differential Scanning Calorimetry (DSC) was conducted using Mettler Toledo's DSC821e. Approximately 2-3 mg of the sample was loaded into the aluminum pan, and its weight was recorded. Petition 870210062456, dated 09 / 07 / 2021, page 26 / 43 16 / 21 precisely. The pan was covered with a lid through which a hole was drilled. The pan was mounted in the equipment and heated from 25 to 250°C at a rate of 10°C / min under nitrogen purge. Indium metal was used as the standard for calibration. The spectra thus obtained are attached to this specification as Figures 3, 7 and 12, respectively. Test Example 4 Thermogravimetry (TG)

[048] Thermogravimetry (TG) was conducted using a Mettler Toledo TGA850. Approximately 4-5 mg of the sample was loaded into the aluminum pan. The pan was mounted on the equipment and then heated from 25 to 250°C at a rate of 10°C / min under nitrogen purge. Nickel and Aluminum™ were used as the standard for calibration. The TG analysis results as obtained are attached to this specification as Figures 3, 7, and 12, respectively, along with the DSC analysis results according to said Test Example 3. Example Test 5 Dynamic vapor adsorption / desorption analysis

[049] Dynamic vapor adsorption / desorption data were collected on the VTI-SA Vapor Sorption Analyzer. While maintained at 25°C, vapor adsorption and desorption were repeated three times at 5% RH intervals in the relative humidity (RH) range of 5~95%. Samples were not dried prior to analysis. The equilibrium standard used for the analysis was that the weight change within 2 minutes is less than 0.01%.

[050] As a result of the analysis, Figure 4 shows a vapor adsorption / desorption isotherm result of crystal form I, from which it can be seen that crystal form I shows a weight change of 0.8% or less according to the change in external humidity in the range of 5~95% RH. That is, crystal form I according to the present Petition 870210062456, dated 09 / 07 / 2021, page 27 / 43 The invention is very stable against changes in relative humidity. Figure 8 shows a vapor adsorption / desorption isotherm result for crystal form II, from which it can be seen that in crystal form II the weight increases by 4.3% when the relative humidity is raised to 95% in the initial mixture adsorption test. After moisture desorption, crystal form II produces the same moisture behavior graph as crystal form I. From this result, it can be confirmed that crystal form II was converted to crystal form I during the course of initial moisture adsorption. Figure 13 shows a vapor adsorption / desorption isotherm result for crystal form III, from which it can be seen that in crystal form III the weight increases by 3.6% when the relative humidity is raised to 95%.Additionally, it can be observed that decreasing the humidity causes moisture desorption of 0.6% or less, so that crystal form III ends up containing the same moisture content as crystal form I. Example Test 6 X-ray diffraction test on crystal form II powder during moisture adsorption.

[051] Approximately 50 mg of crystal form II was placed in a glass vial, placed under relative humidity of 11%, 32%, 53%, 64%, 78% and 97%, respectively, for two or more days to induce moisture adsorption, and then subjected to powder x-ray diffraction testing according to the conditions as presented in said Test Example 1 to identify any change in crystal form at the time of moisture adsorption (see Figure 9).

[052] The respective relative humidity was provided by making the aqueous salt solution saturated as shown in the following Table 3 and placing the solution in a desiccator, which was then sealed. [Table 3] Relative humidity of 11% Saturated aqueous solution of LiCl Petition 870210062456, dated 09 / 07 / 2021, page 28 / 43 18 / 21 Relative humidity of 32% Saturated aqueous solution of MgCU Relative humidity of 53% Saturated aqueous solution of Mg(NO3)2.6H2O Relative humidity of 64% Saturated aqueous solution of NaNO2 Relative humidity of 78% Saturated aqueous solution of NaCl Relative humidity of 97% Saturated aqueous solution of KNO3 Petition 870210062456, dated 09 / 07 / 2021, page 29 / 43 19 / 21 Example Test 7 X-ray diffraction test on powder of crystal form III during moisture adsorption.

[053] Crystal form III was placed in an XRD holder and subjected to the XRD test over time (after 30 minutes, 1 hour, and 5 hours) while allowing moisture adsorption at room temperature under atmosphere (see Figure 15). In addition, the change in weight at room temperature under atmosphere was recorded over time to obtain a graph (see Figure 14). It can be seen that crystal form III rapidly absorbs water so that it is converted into crystal form I within about one hour. Example Test 8 Thermal stability of crystal form I and crystal form II

[054] Approximately 50 mg each of crystal form I and crystal form II were placed in a Duma bottle and then kept at 40±2°C, 75±5% RH or 60±2°C, 5±5% RH. After 2 weeks, 4 weeks, and 8 weeks, each sample was removed from the Duma bottle and analyzed by XRD to identify any change in crystal form and HPLC for stability identification. For HPLC analysis, the sample was dissolved in a mixture of acetonitrile / water / trifluoroacetic acid = 30 / 70 / 0.1 (v / v / v) and then subjected to analysis. The conditions for HPLC analysis are as follows: Conditions for HPLC analysis Loudspeaker: Atlantis dC18 (4.6 mm ID x 250 mm L, Particle size 5 µm, Water) Temperature column: 10°C Mobile phase: Mobile phase A: MeCN / TFA = 100 / 0.1 (v / v) Mobile phase B: H2O / TFA = 100 / 0.1 (v / v) Petition 870210062456, dated 09 / 07 / 2021, pp. 30 / 43 20 / 21 Gradient condition: Time (min) A (%) B (%) Initial 38 62 25 38 62 35 80 20 40 38 62 55 38 62 Flow rate: 0.7 ml / min. Detection: 256 nm, UV Injection volume: 10 μL Total analysis time: 55 min.

[055] The stability results for crystal form I and crystal form II are shown in the following Table 4. [Table 4] Time (weeks) 40°C / 75% RH 60°C / 5% RH Crystal form I Crystal form II Crystal form I Crystal form II 0 99.4 98.4 99.4 98.4 2 99.3 98.4 98.8 98.1 4 99.2 98.3 98.6 97.8 8 99.2 98.2 98.4 97.5

[056] As shown in Table 4, it can be confirmed that maintaining crystal form I and crystal form II at 40±2°C, 75±5% RH or 60±2°C, 5±5% RH exhibits stability greater than 8 weeks. However, according to the XRD analysis result, crystal form I did not show any change up to 8 weeks, but crystal form II was converted to crystal form I at 8 weeks under the condition of 40°C / 75% RH (see Figure 16). Example Test 9 Determination of the density of crystal form I and crystal form II.

[057] Approximately 20-30 ml each of crystal form I and crystal form II were introduced into a cylinder measuring 50 ml. Here, the samples were introduced slowly so that they were not in close contact with each other. After reading a scale on the cylinder of Petition 870210062456, dated 09 / 07 / 2021, pages 31 / 43 21 / 21 measurement, the weight of the sample was measured to calculate the volumetric density. After measuring the volumetric density, the measuring cylinder containing the sample was placed in the ERWEKA densimeter and then subjected to 250 pulses to measure the density (Table 5). The measurement result shows that both the volumetric density and the density of crystal form I were higher than that of crystal form II. The Carr index was calculated from the volumetric density and the density. As a result, it was lower in crystal form I than in crystal form II. As it is generally understood that the lower the Carr index, the better the flowability, it can be identified by comparison to the Carr indices that crystal form I can be easily handled in processing procedures that include tablet preparation compared to crystal form II. [Table 5] Determination of the density and particle size of crystal form I and crystal form II. Form Density Volumetric density Carr Index I 0.45 0.54 16.22 II 0.34 0.44 23.47 Petition 870210062456, dated 09 / 07 / 2021, pages 32 / 43

Claims

1 / 2 CLAIMS 1. Compound, characterized by being a 1,5-hydrated salt of tartrate of 1-{(2S)-2-amino-4-[2,4-bis(trifluoromethyl)-5,8-dihydropyrido[3,4-d]pyrimidin-7(6H)-yl]-4-oxobutyl}-5,5-difluoropiperidin-2-one, and having a water content in the range of 3.5 to 5.5%, wherein the characteristic peak values ​​(2θ) in XRD diffractograms are 14.77±0.1, 15.49±0.1, 17.83±0.1, 20.03±0.1, 21.32±0.1, 22.69±0.1, characteristic absorbance in the infrared (IR) spectrum are 3591, 3401, 3128, 1712, 1655, 1636, 1229, 1205, 1129 and 1059 cm-1; and the differential scanning calorimetry spectrum has endothermic peaks in two temperature ranges of 90-130 °C and 130-160 °C.

2. Process for preparing the 1,5 hydrate as defined in claim 1, characterized in that: tartaric acid is added to 1-{(2S)-2-amino-4-[2,4-bis(trifluoromethyl)-5,8-dihydropyrido[3,4-d]-pyrimidin-7(6H)-yl]-4-oxobutyl}5,5-difluoropiperidin-2-one to obtain the tartrate salt of 1-{(2S)-2-amino-4-[2,4-bis(trifluoromethyl)-5,8-dihydropyrido[3,4-d]-pyrimidin-7(6H)-yl]-4-oxobutyl}-5,5-difluoropiperidin-2-one; and the tartrate salt of 1-{(2S)-2-amino-4-[2,4bis(trifluoromethyl)-5,8-dihydropyrido[3,4-d]-pyrimidin-7(6H)-yl]-4-oxobutyl}5,5-difluoropiperidin-2-one is recrystallized from water, acetonitrile / water, ethanol / water, ethanol / hexane or ethyl acetate / hexane solvents.

3. Process according to claim 2, characterized in that the recrystallization solvent is water.

4. Pharmaceutical composition for inhibiting dipeptidyl peptidase IV, characterized in that it comprises the 1,5 hydrate as defined in claim 1 as the active component together with a pharmaceutically acceptable carrier. Petition 870260057480, dated 12 / 06 / 2026, page 12 / 20 2 / 2 5. Composition according to claim 4, characterized in that it is used for the treatment or prevention of diabetes or obesity. Petition 870260057480, dated 12 / 06 / 2026, page 13 / 20

Images