Method for detecting benzyl bromide analogue impurities in alogliptin benzoate raw material medicine

Abstract

The invention discloses a method for detecting benzyl bromide analogue impurities in an alogliptin benzoate raw material medicine, and relates to the technical field of medicine analysis and detection. According to the method, one-time detection of the three benzyl bromide analogue impurities in the alogliptin benzoate raw material medicine can be achieved, the detection sensitivity can reach the ppm (1 / million) level, and control over potential genotoxic impurities in the medicine can be better and more accurately achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical analysis and detection, and particularly relates to a method for detecting bromobenzyl analog impurities in alogliptin benzoate raw materials. Background Art

[0002] Alogliptin Benzoate, with the chemical name of 2-({6-[(3R)-3-aminopiperidin-1-yl]-3-methyl-2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl}methyl)benzonitrile monobenzoate, has the molecular formula of C 18 H 21 N5O2·C7H6O2, a molecular weight of 461.51, a CAS number of 850649-62-6, and the structural formula is as follows:

[0003]

[0004] Alogliptin Benzoate is suitable for the treatment of type 2 diabetes. When used alone, it is used as an adjunct to diet control and exercise for improving blood glucose control in type 2 diabetes patients. When metformin hydrochloride alone cannot effectively control blood glucose, alogliptin benzoate can be used in combination with metformin hydrochloride to improve blood glucose control in type 2 diabetes patients on the basis of diet and exercise.

[0005] Alogliptin Benzoate is a DPP-4 inhibitor. Eating stimulates the small intestine to secrete incretins with elevated concentrations into the bloodstream, such as glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide. These hormones cause pancreatic β-cells to release insulin in a glucose-dependent manner, but these hormones can be inactivated by DPP-4 enzyme within a few minutes. Alogliptin Benzoate inhibits DPP-4 activity, can slow down the inactivation of these incretins, thereby increasing the blood concentration of these hormones, and reducing the fasting and postprandial blood glucose of type 2 diabetes patients in a glucose-dependent manner. In in vitro studies, when the concentration is close to the therapeutic exposure, alogliptin benzoate selectively binds and inhibits DPP-4 activity, but does not inhibit DPP-8 or DPP-9 activity.

[0006] The preparation route of alogliptin benzoate raw materials is as follows:

[0007]

[0008] The starting material 2-cyanobenzyl bromide used therein contains a halogenated group. According to the current technical requirements, such halogenated groups have potential genotoxicity. In the pharmaceutical process, in order to ensure the safety and effectiveness of drugs, it is necessary to study, detect and monitor some impurities that may have genotoxicity in drug raw materials and their preparations. Due to different synthetic processes of drugs, the impurity profiles of drugs will also change; when the synthetic processes are the same, due to different requirements for the types and contents of impurities between preparations and bulk drugs, or due to different excipients contained in the preparations, the requirements for the monitored impurities of bulk drugs and preparations are also different. Therefore, it is necessary to establish a suitable analytical method according to different synthetic processes and preparation types to achieve accurate and effective detection and monitoring of genotoxic impurities in alogliptin benzoate. Summary of the Invention

[0009] Based on the technical problems existing in the background art, the present invention provides a method for detecting bromobenzyl analog impurities in alogliptin benzoate bulk drug.

[0010] The method for detecting bromobenzyl analog impurities in alogliptin benzoate bulk drug provided by the present invention adopts high performance liquid chromatography, and the chromatographic conditions include: the chromatographic column is Ultimate XB-CN, 250×4.6mm, 5μm; the mobile phase is composed of trifluoroacetic acid aqueous solution and methanol in a volume ratio of 90:10, wherein the pH of the trifluoroacetic acid aqueous solution is 3.4 - 3.8; the flow rate is 0.95 - 1.05ml / min; the detection wavelength is 233 - 237nm.

[0011] Among them, the pH of the trifluoroacetic acid aqueous solution can be, for example, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5; the flow rate can be, for example, 0.95ml / min, 0.96ml / min, 0.97ml / min, 0.98ml / min, 0.99ml / min, 1ml / min, 1.01ml / min, 1.02ml / min, 1.03ml / min, 1.04ml / min, 1.05ml / min; the detection wavelength can be, for example, 233nm, 234nm, 235nm, 236nm, 237nm.

[0012] Preferably, the reagent for adjusting the pH of the trifluoroacetic acid aqueous solution is triethylamine.

[0013] Preferably, in the trifluoroacetic acid aqueous solution, the volume fraction of trifluoroacetic acid is 0.01 - 0.03%.

[0014] Preferably, the column temperature is 30 - 40°C, and can be, for example, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C.

[0015] Preferably, the sample injection volume is 10 - 50 μL, for example, it can be 10 μL, 15 μL, 20 μL, 25 μL, 30 μL, 35 μL, 40 μL, 45 μL, 50 μL.

[0016] Preferably, the length of the chromatographic column is 250 mm, the diameter is 4.6 mm, and the particle size of the packing is 5 μm.

[0017] Preferably, the bromobenzyl analogue impurity is at least one of 2-cyanobenzyl bromide, 3-cyanobenzyl bromide, and 4-cyanobenzyl bromide.

[0018] Among them, the structural formula of 2-cyanobenzyl bromide is:

[0019] The structural formula of 3-cyanobenzyl bromide is:

[0020] The structural formula of 4-cyanobenzyl bromide is:

[0021] The beneficial effects of the present invention are as follows:

[0022] The present invention can achieve the one-time detection of three bromobenzyl analogue impurities in alogliptin benzoate bulk drugs, and its detection sensitivity can reach the ppm (one in a million) level, better and more accurately realizing the control of potential genotoxic impurities in drugs. Description of the Drawings

[0023] Figure 1 It is the liquid chromatogram obtained by the detection method of Example 1 of the present invention. Detailed Description of the Invention

[0024] Next, the technical solutions of the present invention will be described in detail through specific examples.

[0025] Example 1

[0026] The chromatographic conditions are as follows:

[0027] Chromatographic column: Kromasil 60-5CN 250*4.6mm 5μm;

[0028] Mobile phase: Take 20 μL of trifluoroacetic acid and add it to 100 mL of water, adjust the pH value to 3.5 with triethylamine to obtain an aqueous trifluoroacetic acid solution with a volume fraction of 0.02% (pH = 3.5); mix the above-prepared aqueous trifluoroacetic acid solution and methanol evenly according to a volume ratio of 90:10 to obtain the mobile phase;

[0029] Flow rate: 1.0 ml / min; Column temperature: 35 °C; Detection wavelength: 235 nm; Sample injection volume: 20 μL.

[0030] Take 2-cyanobenzyl bromide, 3-cyanobenzyl bromide, and 4-cyanobenzyl bromide respectively, weigh them accurately, dissolve and dilute them with methanol to prepare solutions with a concentration of 1 mg / mL respectively, shake well, and use them as impurity stock solutions; take alogliptin benzoate, weigh it accurately, dissolve and dilute it with the mobile phase to prepare a solution with a concentration of 5 mg / mL, shake well, and use it as the test substance stock solution; accurately measure an appropriate amount of the test substance stock solution and an appropriate amount of the impurity stock solution, and dilute them with the mobile phase to prepare a solution containing 0.25 μg of each of 2-cyanobenzyl bromide, 3-cyanobenzyl bromide, 4-cyanobenzyl bromide, and alogliptin in every 1 mL, and use it as the system solution.

[0031] Accurately measure 20 μL of the system solution, inject it into the liquid chromatograph, and record the chromatogram. The obtained liquid chromatogram is shown in Table 1 and Figure 1 as follows.

[0032] Table 1

[0033]

[0034]

[0035] From the data in Table 1 and Figure 1 it can be seen that under these chromatographic conditions, the resolution between the main component peak and the adjacent impurity peaks is greater than 1.5, and the theoretical plate number is greater than 2000, meeting the requirements, and it can be used as a method for detecting genotoxic impurities of benzyl bromide analog impurities in alogliptin benzoate bulk drug.

[0036] Note: Since 2-cyanobenzyl bromide, 3-cyanobenzyl bromide, and 4-cyanobenzyl bromide have the same elution position under these chromatographic conditions, and 2-cyanobenzyl bromide, 3-cyanobenzyl bromide, and 4-cyanobenzyl bromide are themselves positional isomers, it is acceptable to combine them and detect them as one impurity.

[0037] As mentioned above, it is only the preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art within the technical scope disclosed by the present invention, according to the technical solution and inventive concept of the present invention, makes equivalent substitutions or changes, and should be covered by the protection scope of the present invention.

Claims

1. A method for detecting bromobenzyl analog impurities in alogliptin benzoate bulk drug, characterized in that, The high performance liquid chromatography method is adopted, and the chromatographic conditions include: the chromatographic column is Kromasil 60-5CN 250×4.6 mm, 5 μm; the mobile phase is composed of trifluoroacetic acid aqueous solution and methanol at a volume ratio of 90:10, wherein the pH of the trifluoroacetic acid aqueous solution is 3.4-3.8; the flow rate is 0.95-1.05 ml / min; the detection wavelength is 233-237 nm.

2. The detection method of bromobenzyl analog impurities in alogliptin benzoate bulk drug according to claim 1, characterized in that, The reagent for adjusting the pH of the trifluoroacetic acid aqueous solution is triethylamine.

3. The detection method of bromobenzyl analog impurities in alogliptin benzoate bulk drug according to claim 1, characterized in that, In the trifluoroacetic acid aqueous solution, the volume fraction of trifluoroacetic acid is 0.01-0.03%.

4. The detection method of bromobenzyl analog impurities in alogliptin benzoate bulk drug according to claim 1, characterized in that, The column temperature is 30-40 °C.

5. The detection method of bromobenzyl analog impurities in alogliptin benzoate bulk drug according to claim 1, wherein, The injection volume is 10-50 μL.

6. The detection method of bromobenzyl analog impurities in alogliptin benzoate API according to claim 1, characterized in that, The bromobenzyl analog impurity is at least one of 2-cyanobenzyl bromide, 3-cyanobenzyl bromide, and 4-cyanobenzyl bromide.

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