Method for detecting chloro-uracil impurities in alogliptin

Abstract

The invention provides a method for detecting chloro-uracil impurities in alogliptin, and relates to the technical field of impurity detection. The detection method comprises the step of carrying out component analysis on a solution containing at least one of alogliptin and chloro-uracil impurities shown as I by using a high performance liquid chromatography. According to the detection method disclosed by the invention, six chloro-uracil impurities in alogliptin can be detected at one time, so that efficient detection of potential genotoxic impurities in alogliptin drugs is realized.

Description

Technical Field

[0001] The present invention relates to the technical field of impurity detection, and particularly relates to a method for detecting chlorinated uracil impurities in alogliptin. Background Art

[0002] Alogliptin Benzoate, chemically named 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 C 18 H 21 N5O2·C7H6O2, with 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 applicable to the treatment of type 2 diabetes. When used alone, it is used as an adjuvant treatment to diet control and exercise for improving blood glucose control in type 2 diabetes patients. When hydrochloric acid metformin alone still cannot effectively control blood glucose, alogliptin benzoate can be used in combination with hydrochloric acid metformin to improve blood glucose control in type 2 diabetes patients on the basis of diet and exercise.

[0005] Alogliptin 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 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 level, alogliptin selectively binds and inhibits DPP-4 activity, but does not inhibit DPP-8 or DPP-9 activity.

[0006]

[0007] The above formula is the preparation route of alogliptin benzoate API. The starting material 6-chloro-3-methyluracil 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 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 impurity types and impurity contents in preparations compared with APIs, or due to different excipients contained in preparations, the requirements for the monitored impurities in APIs 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 efficient detection of genotoxic impurities in alogliptin benzoate. Summary of the Invention

[0008] (I) Technical Problems to be Solved

[0009] In view of the deficiencies of the prior art, the present invention provides a method for detecting chlorinated uracil impurities in alogliptin, which solves the technical problem of how to efficiently detect genotoxic impurities in alogliptin.

[0010] (II) Technical Solutions

[0011] To achieve the above object, the present invention is realized through the following technical solutions:

[0012] On the one hand, the present invention provides a method for detecting chlorinated uracil impurities in alogliptin, and the detection method includes performing component analysis on a solution containing at least one of alogliptin and chlorinated uracil impurities shown in formula I by high performance liquid chromatography;

[0013]

[0014] Preferably, the chromatographic conditions of the high performance liquid chromatography include: the chromatographic column is selected from a cyanosilane-bonded silica chromatographic column, and the mobile phase is selected from a trifluoroacetic acid solution - acetonitrile solution.

[0015] Preferably, the chromatographic conditions of the high performance liquid chromatography further include: the flow rate of the mobile phase is 0.95 - 1.0 ml / min, the column temperature of the chromatographic column is 30 - 35 °C, the detection wavelength is 258 - 260 nm, and the injection volume is 10 - 20 μl. For example, the flow rate can be 0.95, 0.96, 0.97, 0.98, 0.99, 1 ml / min, the column temperature can be 30, 31, 32, 33, 34, 35 °C, the detection wavelength can be 258, 259, 260 nm, and the injection volume can be 10, 15, 20 μl.

[0016] Preferably, the pH value of the trifluoroacetic acid solution is 3.0 - 3.2, for example, the pH can be 3.0, 3.1, or 3.2.

[0017] Preferably, the volume ratio of the trifluoroacetic acid solution to acetonitrile is 80 - 85:15 - 20, for example, 80:20, 82:18, or 85:15.

[0018] 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.

[0019] Preferably, the pH of the trifluoroacetic acid solution is adjusted with triethylamine.

[0020] Preferably, the trifluoroacetic acid is an aqueous solution of trifluoroacetic acid.

[0021] Preferably, in the aqueous solution of trifluoroacetic acid, the volume fraction of trifluoroacetic acid is 0.04 - 0.06%.

[0022] Preferably, the chlorinated uracil impurities are at least one of those in the following table, and the specific structural formulas are shown in Table 1 below.

[0023] Table 1

[0024]

[0025] (III) Beneficial Effects

[0026] The present invention provides a method for detecting chlorinated uracil impurities in alogliptin. Compared with the prior art, it has the following beneficial effects:

[0027] The detection method of the present invention can detect 6 kinds of chlorinated uracil impurities in alogliptin at one time, thereby realizing the efficient detection of potential genotoxic impurities in alogliptin drugs. Brief Description of the Drawings

[0028] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0029] Figure 1 It is the high - performance liquid chromatogram in Example 1. Detailed Embodiments

[0030] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described. Apparently, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0031] To better understand the above technical solutions, the above technical solutions will be described in detail below in conjunction with the accompanying drawings of the specification and specific embodiments.

[0032] Preparation Example 1

[0033] Accurately weigh alogliptin benzoate, 6-chloro-3-methyluracil, 5-chlorouracil, 6-chlorouracil, 6-chloro-1-methyluracil, 6-chloro-1,3-dimethyluracil, and 5,6-dichloro-3-methyluracil, and add them to a 0.05% trifluoroacetic acid solution (adjust the pH value to 3.2 with triethylamine)-acetonitrile (85:15) solution to prepare a system solution. In the system solution, the mass concentrations of alogliptin benzoate, 6-chloro-3-methyluracil, 5-chlorouracil, 6-chlorouracil, 6-chloro-1-methyluracil, 6-chloro-1,3-dimethyluracil, and 5,6-dichloro-3-methyluracil are all 10 μg / mL.

[0034] Example 1

[0035] Precisely measure 20 μl of the system solution in Preparation Example 1, inject it into a high-performance liquid chromatograph, and record the chromatogram. The chromatographic conditions of the high-performance liquid chromatography are as follows:

[0036] Chromatographic column: Kromasil 60-5CN 250*4.6mm 5μm;

[0037] Mobile phase: 0.05% trifluoroacetic acid solution (adjust the pH value to 3.2 with triethylamine)-acetonitrile (85:15);

[0038] Flow rate: 1.0 ml / min; Column temperature: 35°C; Detection wavelength: 260 nm; Injection volume: 20 μl.

[0039] The obtained chromatogram is as Figure 1 shown. It can be seen that Figure 1 5 absorption peaks are obtained, and alogliptin benzoate, 6-chloro-3-methyluracil, 5-chlorouracil, 6-chlorouracil, 6-chloro-1-methyluracil, 6-chloro-1,3-dimethyluracil, and 5,6-dichloro-3-methyluracil in the solution are all separated. Moreover, there are few impurity peaks in the chromatogram and the baseline is smooth.

[0040] AfterFigure 1 The obtained spectrogram was further analyzed to obtain the results shown in Table 2. 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. Therefore, under these chromatographic conditions, the detection of potential genotoxic impurities in alogliptin drugs can be achieved.

[0041] Table 2

[0042]

[0043]

[0044] It should be noted that in this article, relational terms such as first and second are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variant thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements not only includes those elements, but also includes other elements not expressly listed, or also includes elements inherent to such process, method, article or device. Without further limitation, an element defined by the statement "including a..." does not exclude the existence of additional identical elements in the process, method, article or device including the said element.

[0045] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

[0046] The present invention uses the above embodiments to illustrate the detailed process flow of the present invention, but the present invention is not limited to the above detailed process flow, that is, it does not mean that the present invention must rely on the above detailed process flow to be implemented. Those skilled in the art should understand that any improvement to the present invention, the equivalent replacement of the raw materials of the products of the present invention, the addition of auxiliary components, the selection of specific methods, etc., all fall within the protection scope and the disclosure scope of the present invention.

Claims

1. A method for detecting chlorinated uracil impurities in alogliptin, characterized in that, The detection method includes performing component analysis on a solution containing at least one of alogliptin and the chlorinated uracil impurities shown in I by high performance liquid chromatography; 2. The detection method according to claim 1, characterized in that, The chromatographic conditions of the high performance liquid chromatography include: the chromatographic column is selected from a cyano-silane bonded silica chromatographic column, and the mobile phase is selected from a trifluoroacetic acid solution - acetonitrile solution.

3. The detection method according to claim 1, wherein The chromatographic conditions of the high performance liquid chromatography further include: the flow rate of the mobile phase is 0.95 - 1.0 ml / min, the column temperature of the chromatographic column is 30 - 35 °C, the detection wavelength is 258 - 260 nm, and the injection volume is 10 - 20 μl.

4. The detection method according to claim 1, wherein The pH value of the trifluoroacetic acid solution is 3.0 - 3.

2.

5. The detection method according to claim 4, characterized in that The pH of the trifluoroacetic acid solution is adjusted with triethylamine.

6. The detection method according to claim 1, characterized in that The mass ratio of the trifluoroacetic acid solution to acetonitrile is 80 - 85:15 - 20.

7. The detection method according to claim 1, characterized in that The length of the chromatographic column is 250 mm, the diameter is 4.6 mm, and the filler particle size is 5 μm.

8. The detection method according to claim 1, wherein The trifluoroacetic acid is an aqueous solution of trifluoroacetic acid.

9. The detection method according to claim 8, wherein In the aqueous solution of trifluoroacetic acid, the volume fraction of trifluoroacetic acid is 0.04 - 0.06%.

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