A crystalline hydrochloride salt of ertugliflozin and a preparation method and application thereof

Abstract

The application discloses a crystalline gemigliptin hydrochloride and a preparation method and application thereof, relates to the technical field of medicinal chemistry, and uses Cu-K alpha radiation; the obtained X-ray powder diffraction pattern has characteristic peaks at 2theta values of 11.43+ / -0.2, 19.10+ / -0.2, 22.77+ / -0.2 and 22.90+ / -0.2; the crystalline gemigliptin hydrochloride is not reported in the prior art, has a rod-like structure, has good bioavailability, and can be applied to the preparation of DPP-IV inhibitors, blood sugar reducing drugs, angiogenesis and regeneration drugs.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical chemistry, and in particular to gemagliptin hydrochloride microcrystals, a preparation method and application thereof. Background Art

[0002] Diabetes and its various complications pose a serious threat to people's health and life. The global incidence and prevalence of diabetes are both showing a significant upward trend. Diabetes is primarily categorized into two types: type 1 and type 2. Type 1 diabetes is caused by the destruction of pancreatic cells, resulting in minimal or no insulin secretion; type 2 diabetes is caused by other factors leading to insulin deficiency and insulin resistance. The prevalence of type 2 diabetes accounts for over 90% of all diabetic patients. Typical complications associated with diabetes include hyperlipidemia, hypertension, retinopathy, and renal insufficiency.

[0003] Thiazolidinediones (stimulating insulin secretion from pancreatic cells), biguanides (inhibiting hepatic glucose production), and α-glucosidase inhibitors (inhibiting intestinal glucose absorption) have been used to treat diabetes. In recent years, peroxisome proliferator-activated receptor γ (PPARγ) agonists (thiazolidinediones, which improve insulin sensitivity) have attracted attention as diabetes treatments. However, these drugs have side effects such as hypoglycemia and weight gain. Therefore, there is an urgent need to develop diabetes treatments with fewer side effects, particularly those that do not cause hypoglycemia and weight gain.

[0004] Numerous studies have found that mice lacking dipeptidyl peptidase-IV (DPP-IV) maintain glucagon-like protein 1 (GLP-1) activity and high insulin levels, leading to lower blood sugar levels. DPP-IV inhibitors can control blood sugar and lipid levels in animal experiments, suggesting that DPP-IV inhibitors could be considered as potential effective drugs for treating diabetes.

[0005] Gemtagliptin is a potent and highly selective DPP-IV inhibitor that has demonstrated excellent therapeutic efficacy for type 2 diabetes. However, existing public information on different salt forms and crystal forms of gemagliptin is scarce. Patents US20060074087A1 and US20060040963A1 only cover derivatives of DPP-IV inhibitors and their synthesis methods, without mentioning any crystal forms or salt forms. Patents US20100274013A1 and US7879848B2, while mentioning gemagliptin hydrochloride and its preparation methods, do not investigate the crystal forms of gemagliptin hydrochloride. Patent US20130203787A1 covers different crystal forms of gemagliptin tartrate and its preparation methods, but does not mention the crystal forms of gemagliptin hydrochloride.

[0006] As is well known, drug polymorphism refers to the formation of different crystal structures and different solid forms of drugs with the same molecular formula under the influence of various factors. Numerous studies have shown that different crystal forms of the same drug can have significant differences in color, hygroscopicity, solubility, dissolution properties, melting point, mechanical properties, stability, etc., which directly affect the efficacy and developability of the drug. In addition, the pulverization, drying, tableting, coating and addition of excipients involved in the preparation production process may cause the drug to undergo crystal transformation. Therefore, the development of a crystal form with stable physical and chemical properties and good bioavailability is of great significance to drug research and development, and can improve the market value and competitiveness of the product.

[0007] In view of the lack of research on gemagliptin hydrochloride microcrystals at home and abroad, the present invention will provide a new type of gemagliptin hydrochloride microcrystals and a preparation process thereof, aiming to lay the foundation for the development and application of a new dosage form of the hypoglycemic drug gemagliptin. Summary of the Invention

[0008] The technical problem to be solved by the present invention is to provide a gemagliptin hydrochloride microcrystal and a preparation method thereof. The gemagliptin hydrochloride microcrystal has a rod-shaped structure and good bioavailability, and can be used to prepare DPP-IV inhibitors, hypoglycemic drugs, and angiogenesis and regeneration drugs.

[0009] The technical problem to be solved by the present invention is achieved by adopting the following technical solutions:

[0010] The first object of the present invention is to provide a gemagliptin hydrochloride microcrystal, which uses Cu-Kα radiation to obtain an X-ray powder diffraction pattern with characteristic peaks at 2θ values ​​of 11.43±0.2°, 19.10±0.2°, 22.77±0.2°, and 22.90±0.2°.

[0011] A second object of the present invention is to provide a method for preparing gemagliptin hydrochloride microcrystals, comprising the following steps:

[0012] (1) using a hydrochloric acid isopropanol solution to carry out amino deprotection and salt formation reaction on the gemagliptin intermediate to obtain a crude gemagliptin hydrochloride;

[0013] (2) Recrystallizing the crude gemagliptin hydrochloride product using ethanol to obtain gemagliptin hydrochloride microcrystals.

[0014] The structural formula of the gemagliptin intermediate is shown below:

[0015]

[0016] Wherein, R is selected from Any one of .

[0017] The third object of the present invention is to provide the use of the gemagliptin hydrochloride microcrystals in the preparation of DPP-IV inhibitors, hypoglycemic drugs, angiogenesis and regeneration drugs.

[0018] The beneficial effects of the present invention are:

[0019] 1. The present invention provides a rod-shaped, bioavailable, and pharmaceutically acceptable gemagliptin hydrochloride microcrystal. Such microcrystals have not been reported in the prior art and can lay an important foundation for the development and application of new dosage forms of gemagliptin.

[0020] 2. The present invention provides a method for preparing gemagliptin hydrochloride microcrystals, which is not only simple to operate, but also has high purity of the obtained gemagliptin hydrochloride, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a hydrogen nuclear magnetic resonance spectrum (H-NMR) of the gemagliptin hydrochloride prepared in the present invention;

[0022] Figure 2 The X-ray powder diffraction (XRPD) pattern of the gemagliptin hydrochloride microcrystals prepared in the present invention;

[0023] Figure 3 This is a thermogravimetric analysis (TGA) diagram of the gemagliptin hydrochloride microcrystals prepared in the present invention;

[0024] Figure 4 This is a high performance liquid chromatography (HPLC) chart of the gemagliptin hydrochloride microcrystals prepared in the present invention;

[0025] Figure 5 This is a confocal microscopic image of the gemagliptin hydrochloride microcrystals prepared in the present invention;

[0026] Figure 6 This is a confocal microscopy image of gemagliptin hydrochloride microcrystals prepared in the comparative example;

[0027] Figure 7 The present invention provides a method for the pharmacokinetic analysis of the gemigliptin hydrochloride microcrystals prepared in the present invention and the gemigliptin hydrochloride microcrystals prepared in the comparative example in mice. DETAILED DESCRIPTION

[0028] In order to make the technical means, creative features, objectives and effects achieved by the present invention easier to understand, the present invention is further described below with reference to specific embodiments and illustrations.

[0029] The invention provides gemagliptin hydrochloride microcrystals. Using Cu-Kα radiation, an X-ray powder diffraction spectrum obtained has characteristic peaks at 2θ values ​​of 11.43±0.2°, 19.10±0.2°, 22.77±0.2°, and 22.90±0.2°.

[0030] Furthermore, the X-ray powder diffraction pattern of the gemagliptin hydrochloride microcrystals also has characteristic peaks at 2θ values ​​of 10.68±0.2°, 17.81±0.2°, 18.07±0.2°, 18.93±0.2°, and 22.16±0.2°.

[0031] Furthermore, the X-ray powder diffraction pattern of the gemigliptin hydrochloride microcrystals also has characteristic peaks at 2θ values ​​of 5.02±0.2°, 5.71±0.2°, 15.71±0.2°, 16.10±0.2°, 21.11±0.2°, 24.26±0.2°, 24.56±0.2°, and 28.65±0.2°.

[0032] Furthermore, the gemagliptin hydrochloride microcrystals are rod-shaped, and their crystal structure is observed using a confocal microscope.

[0033] Furthermore, when the gemagliptin hydrochloride microcrystals are heated to 110°C, the weight loss is 0.71±0.05%, when heated from 110°C to 230°C, the weight loss is 4.46±0.05%; and when heated from 230°C to 503°C, the weight loss is 61.88±0.05%.

[0034] The present invention provides a method for preparing gemagliptin hydrochloride microcrystals, comprising the following steps:

[0035] (1) using a hydrochloric acid isopropanol solution to carry out amino deprotection and salt formation reaction on the gemagliptin intermediate to obtain a crude gemagliptin hydrochloride;

[0036] (2) Recrystallizing the crude gemagliptin hydrochloride product using ethanol to obtain gemagliptin hydrochloride microcrystals.

[0037] The structural formula of the gemagliptin intermediate is shown below:

[0038]

[0039] Wherein, R is selected from Any one of .

[0040] Furthermore, the concentration of hydrogen chloride in the hydrochloric acid isopropanol solution is 1 to 6 mol / L.

[0041] Furthermore, the specific operation of the recrystallization is: heating and dissolving the crude gemagliptin hydrochloride in ethanol, cooling to crystallize, filtering, and drying to obtain gemagliptin hydrochloride microcrystals, wherein the heating and dissolving temperature is 60-80° C.; and the drying is freeze-drying.

[0042] Furthermore, the HPLC purity of the gemagliptin hydrochloride microcrystals is greater than 99.8%, and the maximum single impurity content is less than 0.1%.

[0043] The present invention also provides the use of the gemagliptin hydrochloride microcrystals in preparing DPP-IV inhibitors, hypoglycemic drugs, and angiogenesis and regeneration drugs.

[0044] Example 1

[0045]

[0046] Weigh 20 kg of the gemagliptin intermediate (CAS: 911637-18-8) and dissolve it thoroughly in 100 L of isopropanol. Then, add 20 L of a hydrochloric acid isopropanol solution (hydrogen chloride concentration: 1 mol / L) in portions at 0°C. The reaction is allowed to react at room temperature for 4 hours. Monitor the reaction progress by HPLC. After the complete disappearance of the gemagliptin intermediate, filter the mixture by suction. Rinse the filter cake with isopropanol, collect the solid, and freeze-dry to obtain 16.8 kg of crude gemagliptin hydrochloride.

[0047] 16.8 kg of crude gemigliptin hydrochloride was weighed, and 30 L of ethanol was added. The temperature was raised to 70°C to fully dissolve it, and the mixture was stirred for 1 hour. Then, heating and stirring were stopped, and the mixture was naturally cooled to crystallize. The mixture was filtered and freeze-dried to obtain 15.5 kg of gemigliptin hydrochloride with a yield of 87%. 1 H NMR (400 MHz, CD3OD) δ 5.05-4.92 (m, 2H), 4.04-3.71 (m, 6H), 3.60-3.46 (m, 1H), 3.28-3.10 (m, 2H), 3.08-2.82 (m, 2H), 2.70-2.52 (m, 2H), 2.46-2.29 (m, 2H); see the specific spectrum. Figure 1 .

[0048] The XRPD data of the gemagliptin hydrochloride prepared in Example 1 were measured using an X-ray diffractometer (Bruker D8 ADVANCE). The specific detection parameters are shown in Table 1. The XRPD pattern is shown in Figure 2 .

[0049] Table 1

[0050]

[0051] from Figure 2It can be seen that the XRPD pattern of gemagliptin hydrochloride has characteristic peaks at 2θ values ​​of 19.10±0.2°, 22.90±0.2°, 11.43±0.2°, 22.77±0.2°, 18.93±0.2°, 18.07±0.2°, 17.81±0.2°, 22.16±0.2°, 10.68±0.2°, 16.10±0.2°, 5.02±0.2°, 15.71±0.2°, 21.11±0.2°, 28.65±0.2°, 24.26±0.2°, 5.71±0.2°, and 24.56±0.2°, and the heights of these diffraction peaks are all greater than 10%.

[0052] Thermogravimetric analysis of the gelatin hydrochloride prepared in Example 1 was performed using a TGA / DSC 3+ simultaneous thermal analyzer. The specific test parameters are shown in Table 2, and the TGA diagram is shown in Table 2. Figure 3 .

[0053] Table 2

[0054] project parameter Sample tray 70μl alumina clamp pot Temperature range 25～500℃ Heating rate 10℃ / min Shielding gas Nitrogen Shielding gas flow 20mL / min

[0055] from Figure 3 It can be seen that when gemagliptin hydrochloride is heated to 110°C, the weight loss is 0.71%; when heated from 110°C to 230°C, the weight loss is 4.46%; and when heated from 230°C to 503°C, the weight loss is 61.88%.

[0056] The gelatin hydrochloride prepared in Example 1 was analyzed by HPLC using an Agilent 1260 high performance liquid chromatograph (HPLC). The specific detection parameters are shown in Table 3, and the HPLC spectrum is shown in Figure 4 .

[0057] Table 3

[0058]

[0059]

[0060] from Figure 4 It can be seen that the purity of gemagliptin hydrochloride is 99.83% and the maximum single impurity is 0.05%.

[0061] The microstructure of the gemigliptin hydrochloride prepared in Example 1 was observed using a Nikon A1 HD25 single-photon confocal microscope at a magnification of 40. The confocal microscope image is shown in FIG. Figure 5 .from Figure 5 It can be seen that the gemigliptin hydrochloride prepared in Example 1 has a rod-shaped microcrystalline structure.

[0062] Example 2

[0063]

[0064] 2 kg of the gemagliptin intermediate was weighed and fully dissolved in 10 L of isopropanol. Then, 2 L of a hydrochloric acid isopropanol solution (hydrogen chloride concentration of 1 mol / L) was added portionwise at 0°C and allowed to react at room temperature for 4 h. The reaction progress was monitored by HPLC. After the complete disappearance of the gemagliptin intermediate, the filter cake was filtered and rinsed with isopropanol. The solid was collected and freeze-dried to obtain 1.64 kg of crude gemagliptin hydrochloride.

[0065] 1.64 kg of crude gemigliptin hydrochloride was weighed, and 3 L of ethanol was added. The temperature was raised to 70°C to fully dissolve it, and the mixture was stirred for 1 hour. Then, heating and stirring were stopped, and the mixture was naturally cooled to crystallize. The mixture was filtered and freeze-dried to obtain 1.53 kg of gemigliptin hydrochloride with a yield of 85%.

[0066] Example 3

[0067]

[0068] 2 kg of the gemagliptin intermediate was weighed and fully dissolved in 10 L of isopropanol. Then, 2 L of a hydrochloric acid isopropanol solution (hydrogen chloride concentration of 1 mol / L) was added portionwise at 0°C and allowed to react at room temperature for 4 h. The reaction progress was monitored by HPLC. After the complete disappearance of the gemagliptin intermediate, the filter cake was filtered and rinsed with isopropanol. The solid was collected and freeze-dried to obtain 1.33 kg of crude gemagliptin hydrochloride.

[0069] 1.33 kg of crude gemigliptin hydrochloride was weighed, and 3 L of ethanol was added. The temperature was raised to 70°C to fully dissolve it, and the mixture was stirred for 1 hour. Then, heating and stirring were stopped, and the mixture was naturally cooled to crystallize. The mixture was filtered and freeze-dried to obtain 1.19 kg of gemigliptin hydrochloride with a yield of 83%.

[0070] Example 4

[0071]

[0072] 2 kg of the gemagliptin intermediate was weighed and fully dissolved in 100 L of isopropanol. Then, 2 L of a hydrochloric acid isopropanol solution (hydrogen chloride concentration of 1 mol / L) was added portionwise at 0°C and allowed to react at room temperature for 4 h. The reaction progress was monitored by HPLC. After the complete disappearance of the gemagliptin intermediate, the mixture was filtered, the filter cake was rinsed with isopropanol, and the solid was collected and freeze-dried to obtain 1.50 kg of crude gemagliptin hydrochloride.

[0073] 1.50 kg of crude gemigliptin hydrochloride was weighed, and 30 L of ethanol was added. The temperature was raised to 70°C to fully dissolve it, and the mixture was stirred for 1 hour. Then, heating and stirring were stopped, and the mixture was naturally cooled to crystallize. The mixture was filtered and freeze-dried to obtain 1.37 kg of gemigliptin hydrochloride with a yield of 81%.

[0074] Example 5

[0075]

[0076] 2 kg of the gemagliptin intermediate was weighed and fully dissolved in 100 L of isopropanol. Then, 2 L of a hydrochloric acid isopropanol solution (hydrogen chloride concentration of 1 mol / L) was added portionwise at 0°C and allowed to react at room temperature for 4 h. The reaction progress was monitored by HPLC. After the complete disappearance of the gemagliptin intermediate, the mixture was filtered, the filter cake was rinsed with isopropanol, and the solid was collected and freeze-dried to obtain 1.59 kg of crude gemagliptin hydrochloride.

[0077] 1.59 kg of crude gemigliptin hydrochloride was weighed, and 3 L of ethanol was added. The temperature was raised to 70°C to fully dissolve it, and the mixture was stirred for 1 hour. Then, heating and stirring were stopped, and the mixture was naturally cooled to crystallize. The mixture was filtered and freeze-dried to obtain 1.34 kg of gemigliptin hydrochloride with a yield of 78%.

[0078] Comparative Example

[0079] According to the preparation method of Example 1 of patent CN119285630A.

[0080] Weigh the intermediate of gemagliptin (200g, 0.340mol, CAS:911637-18-8), add 2kg of ethanol to fully dissolve it, and add ceric ammonium nitrate (190g, 0.358mol), and heat to 70°C for reaction. Monitor the reaction progress by HPLC. After the gemagliptin intermediate disappears completely, cool to room temperature, add 2kg of methyl tert-butyl ether and stir to precipitate an orange-red ceric ammonium nitrate solid. Filter, spin-dry the filtrate, and then dissolve it in 600mL of ethanol. Slowly add 340mL of hydrochloric acid aqueous solution (1M), stir at room temperature for 1h, and precipitate a large amount of white solid. Filter, collect the filter cake, and dry to obtain gemagliptin hydrochloride with a yield of 50%.

[0081] The microstructure of the gemigliptin hydrochloride prepared in the comparative example was observed using a Nikon A1 HD25 single-photon confocal microscope at a magnification of 40. The confocal microscope image is shown in Figure 6 .from Figure 6 It can be seen that the gemigliptin hydrochloride prepared in the comparative example has an amorphous microcrystalline structure.

[0082] Pharmacokinetic studies

[0083] BALB / c mice are randomly divided into three groups, including blank group, test group and control group, 5 in each group. Test group and control group are respectively injected with the gemigliptin hydrochloride (concentration is 10 μM, dissolved in 300 μL PBS solution) prepared by the present invention and comparative example through tail vein. At predetermined time point (0, 1, 2, 3, 4, 5, 6, 7, 8, 9h after injection), blood sample is collected from submandibular vein, centrifuged for 15min at 16000rpm rotating speed, obtains plasma. The concentration change of gemigliptin hydrochloride in plasma is measured by HPLC, and according to the ratio of peak area to peak area at 0 o'clock for each sample, the relationship curve of blood drug concentration change over time is drawn.

[0084] from Figure 7 It can be seen that the metabolic clearance rate of the gemigliptin hydrochloride prepared by the present invention in mice is slower than that of the gemigliptin hydrochloride prepared in the comparative example, which indicates that the rod-shaped microcrystalline structure of gemigliptin hydrochloride can exert its blood sugar lowering function more sustainably. Therefore, the gemigliptin hydrochloride prepared by the present invention has great clinical translation value.

[0085] The basic principles, main features, and advantages of the present invention are shown and described above. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The above embodiments and descriptions are merely illustrative of the principles of the present invention. Various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention. Such changes and modifications are intended to fall within the scope of the present invention. The scope of protection claimed in the present invention is defined by the appended claims and their equivalents.

Claims

1. A gemigliptin hydrochloride microcrystal, using Cu-Kα radiation, the obtained X-ray powder diffraction pattern is shown in Figure 2; the gemigliptin hydrochloride microcrystal is rod-shaped.

2. The gemagliptin hydrochloride microcrystals according to claim 1, characterized in that: When the gemagliptin hydrochloride microcrystals are heated to 110° C., the weight loss is 0.71±0.05%. When heated from 110° C. to 230° C., the weight loss is 4.46±0.05%. When heated from 230° C. to 503° C., the weight loss is 61.88±0.05%.

3. The method for preparing gemagliptin hydrochloride microcrystals according to claim 1 or 2, characterized in that: The following steps are involved: (1) using a hydrochloric acid isopropanol solution to carry out amino deprotection and salt formation reaction on the gemagliptin intermediate to obtain a crude gemagliptin hydrochloride; (2) recrystallizing the crude gemagliptin hydrochloride product using ethanol to obtain gemagliptin hydrochloride microcrystals; The structural formula of the gemagliptin intermediate is shown below: Wherein, R is selected from Any one of .

4. The preparation method according to claim 3, wherein: The concentration of hydrogen chloride in the hydrochloric acid isopropanol solution is 1-6 mol / L.

5. The preparation method according to claim 3, wherein: The specific operation of the recrystallization is: heating and dissolving the crude product of gemagliptin hydrochloride in ethanol, cooling and crystallizing, filtering, and drying to obtain gemagliptin hydrochloride microcrystals; the temperature of the heating and dissolving is 60-80° C.; and the drying is freeze-drying.

6. The preparation method according to claim 5, characterized in that: The HPLC purity of the gemagliptin hydrochloride microcrystals is greater than 99.8%, and the maximum single impurity content is less than 0.1%.

7. Use of the gemigliptin hydrochloride microcrystals according to claim 1 or 2 in the preparation of dipeptidyl peptidase-4 inhibitors, hypoglycemic drugs, and angiogenesis and regeneration drugs.

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