Amino acid induced new crystalline form of linagliptin based on protein-drug interaction and method of preparation thereof

Abstract

The application discloses a key amino acid induced new crystal form of linagliptin based on protein-drug interaction structure mining and a preparation method thereof; and the technical scheme is that key amino acids are introduced into a solvent crystallization system of linagliptin to induce five new crystal forms of linagliptin; the preparation method does not need to use a large amount of organic solvents, has the characteristics of being simple, precise and efficient, has strong crystal form regulation capacity, is energy-saving, environment-friendly and safe, and is easy to realize green industrial production.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparation technology, and in particular relates to a method for regulating the crystal form of trelagliptin using key amino acids. Background Technology

[0002] The discovery and regulation of novel drug crystal forms are central to pharmaceutical crystallography research. They provide the foundation for screening advantageous crystal forms with superior physicochemical properties, significant efficacy, and high safety, and are of great value and practical significance to drug crystal form research. Differences in drug crystal forms can affect the efficacy, safety, stability, and formulation process of drugs.

[0003] Crystallization is essentially a molecular assembly process. Intermolecular interactions (such as hydrophobic interactions, hydrogen bonds, van der Waals forces, and electrostatic forces) directly affect molecular assembly, thus influencing the crystallization and crystal structure of substances. Existing studies have reported interactions between trolagliptin and human serum albumin (HSA) and pepsin (PEP), which are intermolecular interactions between drugs and proteins. Based on this interaction, key amino acids with strong interactions with trolagliptin have been identified. Differences in the interactions between these key amino acids and drug molecules alter the assembly of drug molecules, thereby affecting drug crystallization and its crystal structure. Currently, no studies have been reported on drug crystallization induced by key amino acids identified through protein-drug interactions. Furthermore, proteins are essentially formed by the dehydration condensation of amino acids into peptides, and the peptide chains fold and coil to form the three-dimensional structure of proteins. Therefore, delving into the protein interior to identify key amino acids and utilizing their strong interactions with drugs can help study the effects of these key amino acids on the crystallization and crystal form of trolagliptin, precisely inducing new drug crystal forms.

[0004] This study selected key amino acids that play a strong role in the interaction between trolagliptin and human serum albumin (HSA) and trolagliptin and pepsin (PEP) as research subjects. Key amino acids, represented by glutamic acid (GLU), arginine (ARG), alanine (ALA), threonine (THR), and tyrosine (TYR), were studied to explore the formation of new trolagliptin crystal forms induced by key amino acids in protein-drug interactions and their preparation methods.

[0005] Trelagliptin (TRE), chemically named 2-[[6-[(3R)-3-amino-1-piperidinyl]-3,4-dihydro-3-methyl-2,4-dioxo-1(2H)-pyrimidinyl]methyl]-4-fluorobenzonitrile, has the chemical formula C2. 18 H 20FN5O2. Trelagliptin, developed by Takeda Pharmaceutical Company of Japan, is a long-acting dipeptidyl peptidase-4 (DPP-4) inhibitor and a very good oral hypoglycemic agent for the treatment of type 2 diabetes. Its structural formula is shown below.

[0006]

[0007] According to known reports, TRE has various crystalline forms. For example, World Patent WO2014127735-A1 (Invention Title: Crystal form A of trelagliptin useful for eg preparing medicine for treating disease mediated by dipeptidyl peptidase IV, exhibits powder X-ray diffraction pattern, and has diffraction peak at specified angle) discloses a trelagliptin crystal A. The 2θ values ​​of the X-ray powder diffraction pattern of this trelagliptin crystal A are 5.7±0.2°, 11.4±0.2°, 12.5±0.2°, 16.8±0.2°, 17.1±0.2°, 19.4±0.2°, 19.9±0.2°, 20.5±0.2°, 22.5±0.2°, 22.9±0.2°, and 29.1±0.2°. Secondly, Chinese patent CN105384724 (Invention title: A crystalline form of a fluorinated compound and its preparation method) discloses crystalline forms II and IV of trelagliptin and their preparation methods; wherein, the 2θ values ​​of the X-ray powder diffraction patterns of crystalline form II are 20.04°, 20.85°, and 21.86°, and the 2θ values ​​of the X-ray powder diffraction patterns of crystalline form IV are 16.28°, 20.03°, 22.30°, and 27.55°. For example, Chinese patent CN105693691 (Invention title: A new crystalline form of high-purity trelagliptin and its preparation) discloses a method for preparing a new crystalline form of high-purity trelagliptin, the 2θ values ​​of which are 4.8±0.2°, 9.6±0.2°, 18.4±0.2°, and 18.9±0.2° of the X-ray powder diffraction patterns of this new trelagliptin crystalline form. For example, Chinese invention patent CN104003975 (Invention title: New solid form of trelagliptin and its preparation method and uses) discloses five crystalline forms of trelagliptin and their preparation methods, including crystal form A, crystal form B, crystal form C, crystal form D, crystal form E, and an amorphous form. For example, Chinese invention patent (application publication number) CN115785066 (Invention title: New crystal form F of trelagliptin and its preparation method) discloses a method for preparing the new crystal form F of trelagliptin, which uses protein grinding induction to prepare the new crystal form of trelagliptin; the preparation method is green and environmentally friendly.

[0008] However, most of the above methods require stringent conditions and do not utilize safe and environmentally friendly amino acids as regulators for controlling trolagliptin crystallization. This patent, based on the three-dimensional structure of the interactions between trolagliptin and human serum albumin (HSA) and trolagliptin and pepsin (PEP), identifies key amino acids that strongly interact with the drug. It innovatively introduces these key amino acids into the trolagliptin solvent system, inducing the formation of various novel trolagliptin crystal forms and designing their preparation methods. The relevant experiments were conducted using glutamic acid (GLU), phenylalanine (PHE), arginine (ARG), threonine (THR), and tyrosine (TYR) as representative amino acids. Summary of the Invention

[0009] The purpose of this invention is to provide a method for preparing various new crystal forms of trelagliptin by introducing key amino acids into the trelagliptin solvent crystallization system and obtaining them through the interaction between the key amino acids and trelagliptin.

[0010] The second objective of this invention is to provide five new crystal forms of treglitazone.

[0011] Therefore, the first technical solution provided by this invention is as follows:

[0012] A method for preparing novel morphologies of trelagliptin based on protein-drug interactions involves introducing key amino acids into the solvent crystallization system of trelagliptin to induce the formation of various novel morphologies.

[0013] Furthermore, the above-mentioned method for preparing the amino acid-induced new crystal form of trolagliptin based on protein-drug interaction includes the following steps in sequence:

[0014] 1) Add solvent to trelagliptin slowly until trelagliptin is just saturated and dissolved to obtain a trelagliptin solution for later use;

[0015] 2) Dissolve the amino acids in deionized water until the solution is just saturated, thus obtaining an amino acid solution;

[0016] 3) Use a dropper to slowly add the amino acid solution prepared in 2) to the trelagliptin solution prepared in 1), and carry out the whole process in a constant temperature water bath at 25℃-50℃;

[0017] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate, thus inducing trelagliptin to produce a variety of new crystal forms.

[0018] The molar ratio of the amino acid and troglitazone is 1:2 to 1:30.

[0019] Furthermore, in the above-mentioned method for preparing a new polymorph of trelagliptin based on protein-drug interaction, the amino acid is one or any combination of glutamic acid, arginine, threonine, and tyrosine; and the amino acid is one of ethyl acetate, isopropanol, acetone, methanol, ethanol, and acetonitrile.

[0020] The second technical solution provided by the present invention is the above-mentioned amino acid-induced new crystal forms of trelagliptin based on protein-drug interaction, wherein the new crystal forms of trelagliptin are TRE-Ⅰ, TRE-Ⅱ, TRE-Ⅲ, TRE-Ⅳ, TRE-Ⅴ, and TRE-Ⅵ.

[0021] Furthermore, the X-ray powder diffraction patterns of the aforementioned amino acid-induced new crystal form of trelagliptin based on protein-drug interactions, specifically TRE-Ⅰ, have 2θ values ​​of 5.66°±0.2°, 11.35°±0.2°, 12.47°±0.2°, 17.08°±0.2°, 19.88°±0.2°, 22.48°±0.2°, 22.87°±0.2°, and 27.54°±0.2°.

[0022] Furthermore, the 2θ values ​​of the aforementioned amino acid-induced new crystal form of trelagliptin based on protein-drug interaction, TRE-Ⅱ, are 8.07°±0.2°, 13.31°±0.2°, 13.64°±0.2°, 16.29°±0.2°, 16.79°±0.2°, 17.40°±0.2°, 19.40°±0.2°, 21.33°±0.2°, 22.83°±0.2°, 26.26°±0.2°, 26.57°±0.2°, 26.79°±0.2°, 27.49°±0.2°, 27.90°±0.2°, and 33.33°±0.2°.

[0023] Furthermore, the 2θ values ​​of the aforementioned amino acid-induced new crystal form of trelagliptin based on protein-drug interaction, TRE-Ⅲ, are 9.99°±0.2°, 13.29°±0.2°, 15.27°±0.2°, 22.62°±0.2°, 26.79°±0.2°, 27.90°±0.2°, 30.36°±0.2°, and 40.73°±0.2°.

[0024] Furthermore, the 2θ values ​​of the aforementioned amino acid-induced new crystal form of trelagliptin based on protein-drug interaction are 5.47°±0.2°, 11.02°±0.2°, 12.88°±0.2°, 16.59°±0.2°, 20.08°±0.2°, 20.91°±0.2°, 22.17°±0.2°, 23.21°±0.2°, 27.84°±0.2°, and 33.55°±0.2°.

[0025] Furthermore, the 2θ values ​​of the aforementioned amino acid-induced new crystal form of trelagliptin based on protein-drug interaction, TRE-V, are 5.49°±0.2°, 11.04°±0.2°, 12.47°±0.2°, 12.86°±0.2°, 16.59°±0.2°, 20.06°±0.2°, 20.88°±0.2°, 22.19°±0.2°, 28.08°±0.2°, 28.98°±0.2°, 30.47°±0.2°, and 33.54±0.2°.

[0026] Compared with the prior art, the technical solution provided by the present invention has the following technical advantages:

[0027] 1. The technical solution provided by this invention is to innovatively introduce key amino acids into the trelagliptin solvent crystallization system, and combine them with only trace amounts of organic solvents. Through the interaction of different solvent systems, various new crystal forms of trelagliptin and preparation methods are obtained.

[0028] 2. The amino acids in the technical solution provided by this invention are essential components of the human body and are necessary for the synthesis of all proteins in the human body. When combined with drugs, they are not only non-toxic but also meet the requirements for human growth. At the same time, it avoids the use of large amounts of toxic and harmful organic solvents, and not only prepares a new crystal form of trelagliptin, but the preparation process is also precise, environmentally friendly, and safe, in line with the national concept of green production.

[0029] 3. The technical solution provided by this invention is based on the interaction between the key amino acid and trelagliptin, and has obtained five new crystal forms, which has broadened the research on drug crystal forms and provided some guidance for the production of subsequent drug formulations. Attached Figure Description

[0030] Figure 1 The images show the X-ray powder diffraction patterns of five new crystal forms of trelagliptin induced by key amino acids based on protein-drug interaction structure mining, compared with the original crystal form of trelagliptin.

[0031] Figure 2 This is the X-ray powder diffraction pattern of the new crystal form I prepared in Example 1;

[0032] Figure 3 This is the X-ray powder diffraction pattern of the new crystal form II prepared in Example 2;

[0033] Figure 4 This is the X-ray powder diffraction pattern of the new crystal form III prepared in Example 3;

[0034] Figure 5 This is the X-ray powder diffraction pattern of the new crystal form IV prepared in Example 4;

[0035] Figure 6 This is the X-ray powder diffraction pattern of the new crystal form V prepared in Example 5;

[0036] Figure 7 This is a microscopic image of the preparation of the new crystal form I in Example 1;

[0037] Figure 8 These are microscope images of the new crystal form II prepared in Example 2;

[0038] Figure 9 This is a microscopic image of the new crystal form III prepared in Example 3;

[0039] Figure 10 This is an observation diagram of the preparation of the new crystal form IV in Example 4;

[0040] Figure 11 This is an observation image of the preparation of the new crystal form V in Example 5; Detailed Implementation

[0041] The following describes various novel crystal forms of trelagliptin induced by key amino acids in a solvent system and their preparation methods, in conjunction with specific implementation methods and the content of the invention. However, the invention is not limited to these types, and all those related to this patent are protected.

[0042] Example 1

[0043] 1) Accurately weigh 100 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 4 mL of methanol solvent to the Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0044] 2) Take another 6.6 mg of threonine (Thr) and add it to the conical flask. Add 2 mL of deionized water to dissolve it until it is saturated.

[0045] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 37°C.

[0046] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0047] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0048] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scan range was 4–50°, the step size was 0.01313°, and the counting time was 30ms / step. The detection results are as follows: Figure 2 As shown, the 2θ values ​​of the new crystal form TRE-Ⅰ are 5.66°±0.2°, 11.35°±0.2°, 12.47°±0.2°, 17.08°±0.2°, 19.88°±0.2°, 22.48°±0.2°, 22.87°±0.2°, and 27.54°±0.2°. Compared with the existing crystal forms, the differences confirm that it is a new crystal form.

[0049] Example 2

[0050] 1) Accurately weigh 150 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 6 mL of methanol solvent to the Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0051] 2) Take another 10 mg of tyrosine (Tyr) into an Erlenmeyer flask, add 3 mL of 20% hydrochloric acid solution and dissolve until just saturated.

[0052] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 50°C.

[0053] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0054] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0055] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 3 As shown, the 2θ values ​​of the new crystal form TRE-Ⅱ are 8.07°±0.2°, 13.31°±0.2°, 13.64°±0.2°, 16.29°±0.2°, 16.79°±0.2°, 17.40°±0.2°, 19.40°±0.2°, 21.33°±0.2°, 22.83°±0.2°, 26.26°±0.2°, 26.57°±0.2°, 26.79°±0.2°, 27.49°±0.2°, 27.90°±0.2°, and 33.33°±0.2°. Compared with the existing crystal forms, there are differences, confirming it as a new crystal form.

[0056] Example 3

[0057] 1) Accurately weigh 250 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 8 mL of acetonitrile or 6 mL of acetone solvent to the Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0058] 2) Take another 10 mg of tyrosine (Tyr) into an Erlenmeyer flask, add 3 mL of 20% hydrochloric acid solution and dissolve until just saturated.

[0059] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 37°C.

[0060] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0061] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0062] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 4As shown, the 2θ values ​​of the new crystal form TRE-Ⅲ are 9.99°±0.2°, 13.29°±0.2°, 15.27°±0.2°, 22.62°±0.2°, 26.79°±0.2°, 27.90°±0.2°, 30.36°±0.2°, and 40.73°±0.2°. The differences compared to existing crystal forms confirm its status as a new crystal form.

[0063] Example 4

[0064] 1) Accurately weigh 100 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 4 mL of ethyl acetate solvent to the Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0065] 2) Take 3.3 mg of arginine or 2.7 mg of glutamic acid or a mixture of 3.3 mg of arginine and 2.7 mg of glutamic acid, add one of them to an Erlenmeyer flask, and add 2 mL of deionized water to dissolve until just saturated.

[0066] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 25°C.

[0067] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0068] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0069] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 5 As shown, the 2θ values ​​of the new crystal form TRE-Ⅳ are 5.47°±0.2°, 11.02°±0.2°, 12.88°±0.2°, 16.59°±0.2°, 20.08°±0.2°, 20.91°±0.2°, 22.17°±0.2°, 23.21°±0.2°, 27.84°±0.2°, and 33.55°±0.2°. The differences compared to existing crystal forms confirm its status as a new crystal form.

[0070] Example 5

[0071] 1) Accurately weigh 100 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 4.5 mL of isopropanol solvent to the Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0072] 2) Take 3.3 mg of arginine or 2.7 mg of glutamic acid or a mixture of 3.3 mg of arginine and 2.7 mg of glutamic acid, add one of them to an Erlenmeyer flask, and add 2 mL of deionized water to dissolve until just saturated.

[0073] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 40°C.

[0074] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0075] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0076] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 6 As shown, the 2θ values ​​of the new crystal form TRE-V are 5.49°±0.2°, 11.04°±0.2°, 12.47°±0.2°, 12.86°±0.2°, 16.59°±0.2°, 20.06°±0.2°, 20.88°±0.2°, 22.19°±0.2°, 28.08°±0.2°, 28.98°±0.2°, 30.47°±0.2°, and 33.54±0.2°. Compared with the existing crystal forms, there are differences, thus confirming it as a new crystal form.

[0077] Example 6

[0078] 1) Accurately weigh 200 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 6 mL of ethanol solvent to each Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0079] 2) Take another 10 mg of tyrosine (Tyr) into an Erlenmeyer flask, add 3 mL of 20% hydrochloric acid solution and dissolve until just saturated.

[0080] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 40°C.

[0081] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0082] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0083] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 3 As shown, the 2θ values ​​of the new crystal form TRE-Ⅱ are 8.07°±0.2°, 13.31°±0.2°, 13.64°±0.2°, 16.29°±0.2°, 16.79°±0.2°, 17.40°±0.2°, 19.40°±0.2°, 21.33°±0.2°, 22.83°±0.2°, 26.26°±0.2°, 26.57°±0.2°, 26.79°±0.2°, 27.49°±0.2°, 27.90°±0.2°, and 33.33°±0.2°. Compared with the existing crystal forms, there are differences, confirming it as a new crystal form.

[0084] Example 7

[0085] 1) Accurately weigh 100mg of trelagliptin and add it to a 100mL Erlenmeyer flask. Then, add 4mL of acetone to the Erlenmeyer flask at 25℃-50℃ until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0086] 2) Take another 10 mg of tyrosine (TYR) and add it to the conical flask. Add 3 mL of 20% hydrochloric acid to dissolve it until it is saturated.

[0087] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 50°C.

[0088] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0089] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0090] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 4 As shown, the 2θ values ​​of the new crystal form TRE-Ⅲ are 9.99°±0.2°, 13.29°±0.2°, 15.27°±0.2°, 22.62°±0.2°, 26.79°±0.2°, 27.90°±0.2°, 30.36°±0.2°, and 40.73°±0.2°. The differences compared to existing crystal forms confirm its status as a new crystal form.

[0091] Example 8

[0092] 1) Accurately weigh 100 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 5.5 mL of isopropanol solvent to the Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0093] 2) Take 4.9 mg of arginine, or 4.1 mg of glutamic acid, or a mixture of 4.9 mg of arginine and 4.1 mg of glutamic acid, add one of these to an Erlenmeyer flask, and add 3 mL of deionized water to dissolve until just saturated.

[0094] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 40°C.

[0095] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0096] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0097] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 6 As shown, the 2θ values ​​of the new crystal form TRE-V are 5.49°±0.2°, 11.04°±0.2°, 12.47°±0.2°, 12.86°±0.2°, 16.59°±0.2°, 20.06°±0.2°, 20.88°±0.2°, 22.19°±0.2°, 28.08°±0.2°, 28.98°±0.2°, 30.47°±0.2°, and 33.54±0.2°. Compared with the existing crystal forms, there are differences, thus confirming it as a new crystal form.

[0098] Example 9

[0099] 1) Accurately weigh 200 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 5 mL of ethanol solvent to each Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0100] 2) Take another 20 mg of tyrosine (Tyr) into an Erlenmeyer flask, add 6 mL of 20% hydrochloric acid solution and dissolve until just saturated.

[0101] 3) Use a dropper to slowly add the amino acid solution prepared in 2) into the solution prepared in 1), and keep the whole process in a constant temperature water bath at 37°C.

[0102] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0103] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0104] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 3 As shown, the 2θ values ​​of the new crystal form TRE-Ⅱ are 8.07°±0.2°, 13.31°±0.2°, 13.64°±0.2°, 16.29°±0.2°, 16.79°±0.2°, 17.40°±0.2°, 19.40°±0.2°, 21.33°±0.2°, 22.83°±0.2°, 26.26°±0.2°, 26.57°±0.2°, 26.79°±0.2°, 27.49°±0.2°, 27.90°±0.2°, and 33.33°±0.2°. The differences compared to existing crystal forms confirm its status as a new crystal form.

[0105] Example 10

[0106] 1) Accurately weigh 100 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 4.5 mL of isopropanol solvent to the Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0107] 2) Take 10 mg of arginine or 8 mg of glutamic acid or a mixture of 10 mg of arginine and 8 mg of glutamic acid, add one of them to an Erlenmeyer flask, and add 2 mL of deionized water to dissolve until just saturated.

[0108] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 45°C.

[0109] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0110] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0111] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands); Cu Kα radiation (λ = 1.540).

[0112] The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 6As shown, the 2θ values ​​of the new crystal form TRE-V are 5.49°±0.2°, 11.04°±0.2°, 12.47°±0.2°, 12.86°±0.2°, 16.59°±0.2°, 20.06°±0.2°, 20.88°±0.2°, 22.19°±0.2°, 28.08°±0.2°, 28.98°±0.2°, 30.47°±0.2°, and 33.54±0.2°. Compared with the existing crystal forms, there are differences, thus confirming it as a new crystal form.

[0113] Example 11

[0114] 1) Accurately weigh 100 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then add 5.5 mL of ethyl acetate solvent to the Erlenmeyer flask and slowly add it dropwise until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0115] 2) Take 10 mg of arginine or 8 mg of glutamic acid or a mixture of 10 mg of arginine and 8 mg of glutamic acid, add one of them to an Erlenmeyer flask, and add 2 mL of deionized water to dissolve until just saturated.

[0116] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 50°C.

[0117] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0118] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0119] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 5As shown, the 2θ values ​​of the new crystal form TRE-Ⅳ are 5.47°±0.2°, 11.02°±0.2°, 12.88°±0.2°, 16.59°±0.2°, 20.08°±0.2°, 20.91°±0.2°, 22.17°±0.2°, 23.21°±0.2°, 27.84°±0.2°, and 33.55°±0.2°. The differences compared to existing crystal forms confirm its status as a new crystal form.

[0120] Example 12

[0121] 1) Accurately weigh 250 mg of trelagliptin and add it to a 100 mL Erlenmeyer flask. Then, add 6 mL of acetonitrile to the Erlenmeyer flask at 25℃-50℃ until the trelagliptin is just dissolved and saturated. Set aside for later use.

[0122] 2) Take another 12.8 mg of tyrosine (TYR) and add it to the conical flask. Add 6.5 mL of 20% hydrochloric acid to dissolve it until it is saturated.

[0123] 3) Use a dropper to slowly add the amino acid solution prepared in step 2) into the solution prepared in step 1), and keep the whole process in a constant temperature water bath at 50°C.

[0124] 4) After adding the ingredients, seal the container with plastic wrap and make 30-35 holes in the seal. Place it in a fume hood and wait for the crystals to precipitate.

[0125] 5) After crystal precipitation, the crystal growth state was observed and recorded using an inverted microscope. The crystals were separated from the solution by filtration, and the collected crystals were dried in a dry environment at T=30℃. A portion of the obtained crystals was sent for XPRD analysis.

[0126] Record the X-ray powder diffraction pattern of crystalline treglitazone under the following conditions: indoor temperature 25℃, relative humidity <60%; XPert PRO polycrystalline X-ray diffractometer (PANalytical, Netherlands), Cu Kα radiation. The photodiode voltage was 40kV, the tube current was 40mA, the 2θ scanning range was 4–50°, the step size was 0.01313°, the counting time was 30ms / step, and the detection results were as follows: Figure 4 The 2θ values ​​of the new crystal form TRE-Ⅲ are 9.99°±0.2°, 13.29°±0.2°, 15.27°±0.2°, 22.62°±0.2°, 26.79°±0.2°, 27.90°±0.2°, 30.36°±0.2°, and 40.73°±0.2°. The differences compared to existing crystal forms confirm its status as a new crystal form.

[0127] To demonstrate the efficacy of the five new morphologies of trelagliptin provided in this application, the solubility of these five new morphologies and the original morphology in hydrochloric acid solution at pH 1.2, aqueous solution at pH 7, and PBS solutions at pH 6.8 and pH 7.4 were determined below.

[0128] The specific operation is as follows: After the five new crystal forms and the original crystal form reach dissolution equilibrium, samples are taken, filtered through a 0.22 μm filter membrane, and an appropriate amount of filtrate is taken. The absorbance value is measured at a wavelength of 279 nm using ultraviolet-visible spectrophotometry (Chinese Pharmacopoeia 2020 edition). The absorbance is substituted into the standard curve to obtain the solubility of the five new crystal forms and the original crystal form, as shown in Table 1.

[0129] Table 1 Solubility Table

[0130]

[0131] Table 1 shows that the solubility of the five new crystal forms of trelagliptin was significantly higher than that of the original crystal form in hydrochloric acid solution (pH 1.2), deionized water (pH 7.0), sodium phosphate buffer solution (pH 6.8), and sodium phosphate buffer solution (pH 7.4). The high solubility of the new crystal forms can compensate for the poor solubility of the original trelagliptin crystal form.

[0132] The above description is only a specific embodiment of the present invention and not all embodiments. Any equivalent modifications made by other people skilled in the art based on the present invention specification are covered by the claims of the present invention and are also protected by the present invention.

Claims

1. A process for the preparation of a new crystalline form of Trglitazone induced by amino acid based on protein-drug interaction characterized in that, Comprise the following steps in turn: 1) slowly drop solvent into the triglitazone, until the triglitazone is just saturated, to obtain a triglitazone solution for standby; 2) add deionized water or 20% hydrochloric acid solution to the amino acid, just until the amino acid is saturated, to obtain an amino acid solution; 3) then use a rubber bulb dropper to slowly drop the amino acid solution prepared in 2) into the triglitazone solution prepared in 1) in turn, the whole process is carried out in a constant temperature water bath at 25-50 ℃; 4) after the addition is completed, seal with plastic wrap, and pierce 20-35 holes at the sealing position, place in a fume hood to wait for the precipitation of crystals, to obtain a plurality of new crystal forms of induced triglitazone; The molar ratio of the amino acid and the triglitazone is 1:2-1:30; The amino acid is one or any combination of glutamic acid, arginine, threonine, and tyrosine; The triglitazone new crystal form is one of TRE-I, TRE-II, TRE-III, TRE-IV, and TRE-V; The X-ray powder diffraction pattern of the TRE-I has 2θ values of 5.66°±0.2°, 11.35°±0.2°, 12.47°±0.2°, 17.08°±0.2°, 19.88°±0.2°, 22.48°±0.2°, 22.87°±0.2°, and 27.54°±0.2°; The X-ray powder diffraction pattern of the TRE-II has 2θ values of 8.07°±0.2°, 13.31°±0.2°, 13.64°±0.2°, 16.29°±0.2°, 16.79°±0.2°, 17.40°±0.2°, 19.40°±0.2°, 21.33°±0.2°, 22.83°±0.2°, 26.26°±0.2°, 26.57°±0.2°, 26.79°±0.2°, 27.49°±0.2°, 27.90°±0.2°, and 33.33°±0.2°; The X-ray powder diffraction pattern of the TRE-III has 2θ values of 9.99°±0.2°, 13.29°±0.2°, 15.27°±0.2°, 22.62°±0.2°, 26.79°±0.2°, 27.90°±0.2°, 30.36°±0.2°, and 40.73°±0.2°; The X-ray powder diffraction pattern of the TRE-IV has 2θ values of 5.47°±0.2°, 11.02°±0.2°, 12.88°±0.2°, 16.59°±0.2°, 20.08°±0.2°, 20.91°±0.2°, 22.17°±0.2°, 23.21°±0.2°, 27.84°±0.2°, and 33.55°±0.2°; The X-ray powder diffraction pattern 2θ value of the TRE-V is 5.49°±0.2°, 11.04°±0.2°, 12.47°±0.2°, 12.86°±0.2°, 16.59°±0.2°, 20.06°±0.2°, 20.88°±0.2°, 22.19°±0.2°, 28.08°±0.2°, 28.98°±0.2°, 30.47°±0.2°, 33.54°±0.2°.

2. A process for the preparation of a new crystalline form of trigliptin induced by amino acids based on protein-drug interaction as claimed in claim 1, wherein, The solvent for dissolving the linagliptin is one of ethyl acetate, isopropyl alcohol, acetone, methanol, ethanol, acetonitrile.

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