Gemigliptin oxalate as well as crystal form, preparation method and application thereof

Abstract

The invention discloses a gemigliptin oxalate and a crystal form, a preparation method and application thereof, relates to the technical field of medicinal chemistry, and provides the crystal form of the gemigliptin oxalate, the crystal form is not reported in the prior art, and animal experiments show that compared with gemigliptin tartrate and gemigliptin hydrochloride, the crystal form of the gemigliptin oxalate has the advantages that the crystal form of the gemigliptin oxalate is more stable. The gemigliptin oxalate prepared by the invention has better pharmacokinetic performance, and lays a foundation for development and application of a new dosage form of gemigliptin; the invention further provides a simple and efficient gemigliptin salt forming process, compared with a method for removing an amino protecting group through acid treatment, the salt forming process provided by the invention does not need to add alkali to dissociate the amino group, impurity generation caused by strong acid or strong alkali is avoided, and the prepared gemigliptin oxalate is high in purity.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical chemistry, and in particular to gemagliptin oxalate and its crystal form, preparation method and application. Background Art

[0002] Diabetes is a chronic metabolic disease caused by the pancreas' inability to produce enough insulin or the body's inability to effectively use insulin, which in turn leads to elevated blood sugar levels. Long-term high blood sugar levels can seriously damage organs such as the heart, blood vessels, kidneys, nerves, and eyes. The latest statistics from the World Health Organization show that approximately 422 million people worldwide suffer from diabetes, of which approximately 90% suffer from type 2 diabetes mellitus (T2DM). Type 2 diabetes is characterized by the gradual deterioration of pancreatic β-cell function, and drug intervention has become an important part of the treatment of type 2 diabetes.

[0003] Gelagliptin is a potent and highly selective dipeptidyl peptidase 4 (DPP-4) inhibitor, launched by LG Life Sciences Co., Ltd. of South Korea in 2012 under the brand name "Zemiglo". Compared with other oral antidiabetic drugs, whether it is monotherapy or combined with other oral hypoglycemic drugs, Gelagliptin has obvious advantages in the treatment of patients with type 2 diabetes. For example, in clinical practice, Gelagliptin has a better protective tendency against cardiovascular disease and kidneys, and in terms of drug combination, Gelagliptin requires a lower dose when used in combination with metformin. In addition, Gelagliptin is more effective than other oral antidiabetic drugs in reducing glycated hemoglobin (HbA1c) and improving pancreatic β-cell function (HOMA-β).

[0004] However, the active ingredients of a drug can exist in the form of different salts. Different crystal forms of the same drug molecule can have significant differences in color, solubility, dissolution performance, melting point, mechanical properties, stability, etc., which directly affect the efficacy and developability of the drug. A Korean patent document with publication number KR1020220048952A discloses an oral compound for the treatment of type 2 diabetes, involving a tartrate salt of gemagliptin, but the invention does not mention the crystal form of the gemagliptin salt, nor does it analyze and study the crystal form of the pharmaceutically acceptable salt of gemagliptin. An Australian patent document with publication number AU2013330679A8 discloses a compound drug composed of gemagliptin tartrate and metformin as active ingredients and a method for preparing the same. Similarly, the patent does not analyze and study any salt crystal form of gemagliptin. At present, the research reports on gemagliptin mainly involve the tartrate and hydrochloride of gemagliptin, and there are few research and reports on gemagliptin oxalate, and there is no research and report on the crystal form of gemagliptin oxalate. Therefore, the present invention provides a new and universal gemagliptin salt-forming process, prepares gemagliptin oxalate, and studies this new crystal form, which lays a foundation for the development and application of a new dosage form of a hypoglycemic drug - gemagliptin. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a method for preparing a pharmaceutically acceptable gemagliptin oxalate, by which the gemagliptin oxalate and its crystal forms which have not been reported so far can be obtained, and a gemagliptin salt-forming process can also be provided based on the preparation method, so as to be suitable for the preparation of other salt forms of gemagliptin.

[0006] The technical problem to be solved by the present invention is achieved by adopting the following technical solutions:

[0007] The first object of the present invention is to provide a gemagliptin oxalate, the structural formula of which is shown below:

[0008]

[0009] The second object of the present invention is to provide a method for preparing gemagliptin oxalate, comprising the following steps:

[0010] (1) using ammonium cerium nitrate to deprotect the amino group of the gemagliptin intermediate to obtain gemagliptin;

[0011] (2) Gemtagliptin reacts with oxalic acid to form a salt to obtain Gemtagliptin oxalate.

[0012] The structural formula of the gemagliptin intermediate is shown below:

[0013]

[0014] Among them, R is selected from Any one of .

[0015] The third object of the present invention is to provide a crystalline form of gemagliptin oxalate, whose X-ray powder diffraction spectrum has characteristic peaks at 2θ values ​​of 24.71±0.2°, 15.63±0.2°, and 19.69±0.2°.

[0016] The fourth object of the present invention is to provide the use of the gemagliptin oxalate in the preparation of dipeptidyl peptidase-44 (DPP-4) inhibitors, hypoglycemic drugs, angiogenesis and regeneration drugs.

[0017] A fifth object of the present invention is to provide a process for forming a gemagliptin salt, comprising the following steps:

[0018] (1) using ammonium cerium nitrate to deprotect the amino group of the gemagliptin intermediate to obtain gemagliptin;

[0019] (2) Gemigliptin reacts with an acid to form a salt to obtain a gelatin salt.

[0020] The structural formula of the gemagliptin intermediate is shown below:

[0021]

[0022] Among them, R is selected from Any one of .

[0023] The beneficial effects of the present invention are:

[0024] 1. The present invention provides a crystalline form of gemagliptin oxalate, which has not been reported in the prior art and lays a foundation for the development and application of new dosage forms of gemagliptin.

[0025] 2. The present invention provides a simple and efficient gemagliptin oxalate formation process. Compared with the method of removing the amino protecting group by acid treatment, the salt formation process provided by the present invention does not need to add alkali to liberate the amino group, thus avoiding the generation of impurities caused by strong acid or strong base, and the prepared gemagliptin oxalate has high purity. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The H-NMR spectrum of the gemagliptin oxalate crystal prepared by the present invention;

[0027] Figure 2 The X-ray powder diffraction (XRPD) pattern of the gemagliptin oxalate crystalline form prepared by the present invention;

[0028] Figure 3 The thermogravimetric analysis (TGA) diagram of the gemagliptin oxalate crystal prepared by the present invention;

[0029] Figure 4 The high performance liquid chromatography (HPLC) diagram of the gemagliptin oxalate crystal prepared by the present invention;

[0030] Figure 5 The high performance liquid chromatography (HPLC) chart of gemagliptin hydrochloride prepared by the present invention;

[0031] Figure 6 The high performance liquid chromatography (HPLC) chart of gemagliptin tartrate prepared by the present invention;

[0032] Figure 7 The present invention provides the in vivo pharmacokinetic curves of gemagliptin oxalate, gemagliptin hydrochloride and gemagliptin tartrate in mice. DETAILED DESCRIPTION

[0033] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further described below in conjunction with specific embodiments and diagrams.

[0034] The present invention provides a gemagliptin oxalate, the structural formula of which is shown below:

[0035]

[0036] The present invention provides a method for preparing gemagliptin oxalate, comprising the following steps:

[0037] (1) using ammonium cerium nitrate to deprotect the amino group of the gemagliptin intermediate to obtain gemagliptin;

[0038] (2) Gemtagliptin reacts with oxalic acid to form a salt to obtain Gemtagliptin oxalate.

[0039] The structural formula of the gemagliptin intermediate is shown below:

[0040]

[0041] Among them, R is selected from Any one of .

[0042] In some specific embodiments, the HPLC purity of the gemagliptin oxalate is greater than 99.4%, and the maximum single impurity content is less than 0.2%.

[0043] The invention provides a gemagliptin oxalate crystal form, and an X-ray powder diffraction spectrum of the crystal form has characteristic peaks at 2θ values ​​of 24.71±0.2°, 15.63±0.2° and 19.69±0.2°.

[0044] In some specific embodiments, the X-ray powder diffraction pattern of the gemagliptin oxalate crystalline form has characteristic peaks at 2θ values ​​of 10.38±0.2°, 15.63±0.2°, 17.28±0.2°, 19.69±0.2°, 22.20±0.2°, 23.13±0.2°, and 24.71±0.2°.

[0045] In some specific embodiments, the X-ray powder diffraction pattern of the gemagliptin oxalate crystalline form has characteristic peaks at 2θ values ​​of 10.38±0.2°, 15.63±0.2°, 16.61±0.2°, 17.28±0.2°, 19.69±0.2°, 20.54±0.2°, 22.20±0.2°, 23.13±0.2°, 24.71±0.2°, 29.88±0.2°, and 35.15±0.2°.

[0046] Furthermore, the X-ray powder diffraction pattern of the gemagliptin oxalate crystalline form is measured using Cu-Kα radiation.

[0047] Furthermore, when the gemagliptin oxalate crystal is heated to 90°C, the weight loss is 0.77%; when heated from 90°C to 136°C, the weight loss is 3.01%; when heated from 136°C to 173°C, the weight loss is 3.50%; and when heated from 173°C to 503°C, the weight loss is 62.57%.

[0048] The present invention provides the use of the gemagliptin oxalate in the preparation of a dipeptidyl peptidase-44 (DPP-4) inhibitor, a hypoglycemic drug, and angiogenesis and regeneration drugs.

[0049] The present invention provides a gemagliptin salt-forming process, comprising the following steps:

[0050] (1) using ammonium cerium nitrate to deprotect the amino group of the gemagliptin intermediate to obtain gemagliptin;

[0051] (2) Gemigliptin reacts with an acid to form a salt to obtain a gelatin salt.

[0052] The structural formula of the gemagliptin intermediate is shown below:

[0053]

[0054] Among them, R is selected from Any one of .

[0055] Furthermore, the acid is an inorganic acid or an organic acid. The inorganic acid includes but is not limited to one of hydrochloric acid and sulfuric acid. The organic acid includes but is not limited to one of tartaric acid, oxalic acid, malic acid, fumaric acid, benzenesulfonic acid, benzoic acid, hippuric acid, adipic acid, succinic acid, and maleic acid.

[0056] Furthermore, the gemagliptin salt includes but is not limited to one of gemagliptin hydrochloride, gemagliptin sulfate, gemagliptin tartrate, gemagliptin oxalate, gemagliptin malate, gemagliptin fumarate, gemagliptin besylate, gemagliptin benzoate, gemagliptin hippurate, gemagliptin adipate, gemagliptin succinate, and gemagliptin maleate.

[0057] Example 1

[0058] Synthesis of Gemagliptin Oxalate:

[0059]

[0060] Weigh 100g (Compound II, CAS: 911637-18-8, 0.170mol) in a reaction bottle, add 1kg ethanol to fully dissolve, add 95g (0.179mol) cerium ammonium nitrate at room temperature, and heat to 70°C for reaction. Monitor the reaction progress by HPLC, cool to room temperature after the raw material disappears, add 1kg methyl tert-butyl ether and stir thoroughly, precipitate orange-red cerium ammonium nitrate solid, filter and spin-dry the filtrate to obtain the crude product of Compound I. Dissolve the crude product of Compound I in 300mL ethanol, slowly drop 50mL of an aqueous solution containing 15g (0.170mol) oxalic acid, heat to 70°C for reaction for 3h, cool to room temperature and continue stirring overnight to react, precipitate a large amount of white solid, filter, collect the filter cake, dry, and obtain a white solid, i.e., Compound III, with a yield of 60%.

[0061] H-NMR data of compound III: 1 H NMR (400MHz, D2O) δ5.01-4.89 (m, 2H), 4.12-3.68 (m, 6H), 3.63-3.52 (m, 1H), 3.34-3.13 (m, 2H), 3.10-2.86 (m, 2H), 2.73-2.52 (m, 2H), 2.46-2.23 (m, 2H); see for specific spectra Figure 1 .

[0062] The XRPD data of the compound III sample prepared in Example 1 was measured by a Bruker D8 ADVANCE analyzer. The detection parameters are shown in Table 1. The XRPD spectrum is shown in Figure 2 .

[0063] Table 1 XRPD test parameters of gemagliptin oxalate

[0064]

[0065] from Figure 2It can be seen that the XRPD spectrum of the gemigliptin oxalate crystalline form has characteristic peaks at 2θ values ​​of 24.71±0.2°, 15.63±0.2°, 19.69±0.2°, 10.38±0.2°, 23.13±0.2°, 17.28±0.2°, 22.20±0.2°, 29.88±0.2°, 20.54±0.2°, 35.15±0.2°, and 16.61±0.2°, and the height % of these diffraction peaks are all greater than 10.

[0066] Thermogravimetric analysis (TGA) data of the compound III sample prepared in Example 1 was collected by TGA / DSC 3+. The specific experimental parameters are shown in Table 2. The TGA graph is shown in Figure 3 .

[0067] Table 2 TGA test parameters of gemagliptin oxalate

[0068] project parameter Sample tray Alumina ceramics Temperature range 30-500℃ Heating rate 10K / min Protective gas Nitrogen Shielding gas flow 20mL / min

[0069] from Figure 3 It can be seen that when the gemagliptin oxalate crystal is heated to 90°C, the weight loss is 0.77%; when heated from 90°C to 136°C, the weight loss is 3.01%; when heated from 136°C to 173°C, the weight loss is 3.50%; when heated from 173°C to 503°C, the weight loss is 62.57%.

[0070] The high performance liquid chromatography (HPLC) data of the compound III sample prepared in Example 1 was collected on an Agilent 1260HPLC. The specific instrument and experimental parameters are shown in Table 3. The HPLC spectrum is shown in Figure 4 .

[0071] Table 3 HPLC test parameters of gemagliptin oxalate

[0072] project parameter Chromatographic columns Hypeisil ODS2 4.6*150mm,5μm Detection wavelength 210nm Injection volume 2μL Flow rate 1ml / min Elution method Gradient elution Column temperature 35℃ Mobile phase Acetonitrile / water=50:50 (V / V) time 30min

[0073] from Figure 4 It can be seen that the purity of gemagliptin oxalate is 99.49% and the maximum single impurity is 0.19%.

[0074] Example 2

[0075] Synthesis of Gemagliptin Hydrochloride:

[0076]

[0077] Weigh 100g (Compound II, CAS: 911637-18-8, 0.170mol) in a reaction bottle, add 1kg ethanol to fully dissolve, add 95g (0.179mol) cerium ammonium nitrate at room temperature, and heat to 70°C for reaction. Monitor the reaction progress by HPLC, cool to room temperature after the raw material disappears, add 1kg methyl tert-butyl ether and stir thoroughly, precipitate orange-red cerium ammonium nitrate solid, filter and spin-dry the filtrate to obtain the crude product of Compound I. Dissolve the crude product of Compound I in 300mL ethanol, slowly add 170ml (1M, 0.170mol) hydrochloric acid aqueous solution at room temperature, continue stirring and reacting at room temperature for 1h, precipitate a large amount of white solid, filter, collect the filter cake, dry, and obtain a white solid, i.e., Compound IV, with a yield of 50%.

[0078] The high performance liquid chromatography (HPLC) data of the compound IV sample prepared in Example 2 was collected on an Agilent 1260 HPLC. The specific instrument and experimental parameters are shown in Table 3. The HPLC spectrum is shown in Figure 5 .

[0079] from Figure 5 It can be seen that the purity of gemagliptin hydrochloride is 98.33% and the maximum single impurity is 0.73%.

[0080] Example 3

[0081] Synthesis of Gemtagliptin Tartrate:

[0082]

[0083] Weigh 100g (Compound II, CAS: 911637-18-8, 0.170mol) in a reaction bottle, add 1kg ethanol to fully dissolve, add 95g (0.179mol) cerium ammonium nitrate at room temperature, and heat to 70°C for reaction. Monitor the reaction progress by HPLC, cool to room temperature after the raw material disappears, add 1kg methyl tert-butyl ether and stir thoroughly, precipitate orange-red cerium ammonium nitrate solid, filter and spin-dry the filtrate to obtain the crude product of Compound I. Dissolve the crude product of Compound I in 300mL ethanol, slowly drop 50mL of an aqueous solution containing 26g (0.170mol) oxalic acid, heat to 70°C for reaction for 3h, cool to room temperature and continue stirring overnight to react, precipitate a large amount of white solid, filter, collect the filter cake, dry, and obtain a white solid, i.e., Compound V, with a yield of 57%.

[0084] The high performance liquid chromatography (HPLC) data of the compound V sample prepared in Example 3 were collected on an Agilent 1260HPLC. The specific instrument and experimental parameters are shown in 3. The HPLC spectrum is shown in Figure 6 .

[0085] from Figure 6It can be seen that the purity of gemagliptin tartrate is 99.65% and the maximum single impurity is 0.09%.

[0086] Pharmacokinetic studies

[0087] BALB / c mice (purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd., 5-6 weeks old, weighing about 20g) were randomly divided into three groups, 5 mice in each group, and the three groups of mice were injected with PBS solutions of gemigliptin tartrate, gemigliptin oxalate, and gemigliptin hydrochloride through the tail vein, respectively, with a concentration of 10μM. Blood samples were collected from the submandibular vein at predetermined time points (0, 1, 2, 3, 4, 5, 6, 7, 8, and 9h after injection), and the samples were centrifuged at 16000r / min for 15min to obtain plasma. The concentration change of gemigliptin salt in plasma was determined by HPLC, and the relationship curve of blood drug concentration over time was drawn according to the ratio of the peak area of ​​each sample to the peak area at 0.

[0088] Figure 7 The pharmacokinetic curves of gemagliptin oxalate, gemagliptin tartrate and gemagliptin hydrochloride prepared by the present invention in mice (tail vein injection) are shown. Figure 7 It can be seen that the metabolic clearance rate of the gemigliptin oxalate prepared by the present invention in mice is slower than that of gemigliptin tartrate and gemigliptin hydrochloride, which indicates that gemigliptin oxalate can exert its blood sugar lowering function more sustainably. Therefore, the gemigliptin oxalate prepared by the present invention has great clinical transformation value.

[0089] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A gemagliptin oxalate, characterized in that: The structural formula of the gemagliptin oxalate is shown below:

2. The method for preparing gemagliptin oxalate according to claim 1, characterized in that: The steps include: (1) using ammonium cerium nitrate to deprotect the amino group of the gemagliptin intermediate to obtain gemagliptin; (2) gemagliptin reacts with oxalic acid to form a salt to obtain gemagliptin oxalate; The structural formula of the gemagliptin intermediate is shown below: Among them, R is selected from Any one of .

3. Use of the gemagliptin oxalate according to claim 1 or the gemagliptin oxalate prepared by the preparation method according to claim 2 in the preparation of dipeptidyl peptidase-44 inhibitors, hypoglycemic drugs, angiogenesis and regeneration drugs.

4. A crystalline form of gemagliptin oxalate, characterized in that: The X-ray powder diffraction spectrum of the gemagliptin oxalate crystal has characteristic peaks at 2θ values ​​of 24.71±0.2°, 15.63±0.2°, and 19.69±0.2°.

5. The gemagliptin oxalate crystalline form according to claim 4, characterized in that: The X-ray powder diffraction spectrum of the gemagliptin oxalate crystalline form has characteristic peaks at 2θ values ​​of 10.38±0.2°, 15.63±0.2°, 17.28±0.2°, 19.69±0.2°, 22.20±0.2°, 23.13±0.2°, and 24.71±0.2°.

6. The gemagliptin oxalate crystalline form according to claim 4, characterized in that: The X-ray powder diffraction pattern of the gemagliptin oxalate crystalline form has characteristic peaks at 2θ values ​​of 10.38±0.2°, 15.63±0.2°, 16.61±0.2°, 17.28±0.2°, 19.69±0.2°, 20.54±0.2°, 22.20±0.2°, 23.13±0.2°, 24.71±0.2°, 29.88±0.2°, and 35.15±0.2°.

7. The gemagliptin oxalate crystalline form according to any one of claims 4 to 6, characterized in that: The X-ray powder diffraction pattern of the gemagliptin oxalate crystalline form is measured using Cu-Kα radiation.

8. The gemagliptin oxalate crystalline form according to any one of claims 4 to 6, characterized in that: The gemagliptin oxalate crystal loses 0.77% of its weight when heated to 90°C; 3.01% when heated from 90°C to 136°C; 3.50% when heated from 136°C to 173°C; and 62.57% when heated from 173°C to 503°C.

9. A process for forming a salt of gemagliptin, characterized in that: The steps include: (1) using ammonium cerium nitrate to deprotect the amino group of the gemagliptin intermediate to obtain gemagliptin; (2) gemagliptin reacts with an acid to form a salt to obtain a gemagliptin salt; The structural formula of the gemagliptin intermediate is shown below: Among them, R is selected from Any one of .

10. The process for forming the gemagliptin salt according to claim 9, characterized in that: The acid is an inorganic acid or an organic acid; the inorganic acid is one of hydrochloric acid and sulfuric acid; the organic acid is one of tartaric acid, oxalic acid, malic acid, fumaric acid, benzenesulfonic acid, benzoic acid, hippuric acid, adipic acid, succinic acid, and maleic acid; The gemagliptin salt is one of gemagliptin hydrochloride, gemagliptin sulfate, gemagliptin tartrate, gemagliptin oxalate, gemagliptin malate, gemagliptin fumarate, gemagliptin besylate, gemagliptin benzoate, gemagliptin hippurate, gemagliptin adipate, gemagliptin succinate, and gemagliptin maleate.

Images