Detection method for related substances in fumaric acid Vonoprasone intermediate product

Abstract

The invention discloses a method for detecting related substances of a fumaric acid Vonoprasone intermediate product by applying a high performance liquid chromatography, and the chromatographic conditions are as follows: a chromatographic column filler uses high-purity porous spherical silica gel particles coated with charged nano polymer microspheres, an ammonium acetate-acetonitrile system is used as a mobile phase, an ultraviolet detector is used as a detector, and the chromatographic column filler is used for detecting the related substances of the fumaric acid Vonoprasone intermediate product. According to the method, the detection of intermediate products 1 related substances, namely 4-dimethylaminopyridine and pyridine-3-sulfonic acid, generated in the production process of the Vonorazan fumarate is accurately realized, and a simple and reliable method is provided for the detection of the intermediate products generated in the production process of the Vonorazan fumarate.

Description

Technical Field

[0001] The invention relates to the field of pharmaceutical technology, specifically to a detection method for a novel oral potassium ion competitive acid blocker (P-CAB) vonoprazan fumarate, and particularly to a detection method for related substances of a vonoprazan fumarate intermediate. Background Art

[0002] Vonoprazan fumarate (TAK-438) is a novel oral potassium ion competitive acid blocker (P-CAB), whose chemical name is 1-[5-(2-fluorophenyl)-1-(pyridine-3-sulfonyl)-1H-pyrrol-3-yl]-N-methylmethylamine monofumarate, which can effectively inhibit the formation of upper gastrointestinal mucosal damage and is used to treat reflux esophagitis and to eradicate Helicobacter pylori in combination with appropriate antibiotics. The structure of vonoprazan fumarate is shown in the following formula.

[0003] .

[0004] According to reports in many literatures (CN105143187A and CN113121399A of patent applications of the original research company Takeda, J.Med.Chem.2012,55,4446-4456), the main synthetic routes of vonoprazan fumarate are as follows: Figure 1 As shown: 3-pyridinesulfonyl chloride (SM2) and 5-(2-fluorophenyl)-1H-pyrrole-3-carboxaldehyde (SM1) are subjected to condensation reaction to obtain -(2-fluorophenyl)-1-[(pyridin-3-yl)sulfonyl]-1H-pyrrole-3-carboxaldehyde (intermediate product 1), which is then subjected to amination reduction to form a salt to obtain vonoprazan fumarate.

[0005] In the process of preparing the intermediate product 1, acetonitrile is usually used as a solvent, triethylamine is used as an acidifying agent, and a catalyst 4-dimethylaminopyridine is added to carry out a condensation reaction to obtain the intermediate product 1. The structure of the intermediate product 1 is shown in the following formula.

[0006] .

[0007] In the process of preparing intermediate product 1, pyridine-3-sulfonic acid and 4-dimethylaminopyridine are two main impurities, and their structures are shown in the following formula 2.

[0008] .

[0009] The detection methods for related substances in vonoprazan fumarate intermediates are not recorded in the Pharmacopoeia of the People's Republic of China (CHP2020), the European Pharmacopoeia (EP), the United States Pharmacopoeia (USP40) and the Japanese Pharmacopoeia (JP17).

[0010] At present, the literature on the use of high performance liquid chromatography for the detection of vonoprazan fumarate related substances includes: Chinese Journal of Pharmaceutical Affairs. 2022, 36 (07), 800-809 - using a ZORBZX SB-C18 column (4.6 mm × 150 mm, 1.8 μm), mobile phase A is 0.025 mol·L -1 Phosphate buffer solution (pH 6.0) methanol-acetonitrile (14:5:1), mobile phase B was 0.025 mol·L -1 Phosphate buffer solution (pH 6.0)-acetonitrile (3:7) was used to detect 14 impurities in vonoprazan fumarate; China Pharmaceutical Industry. 2023, 32 (11), 63-67——Using Agilent 5 TC-C 18 The z column (250mm×4.6mm, 2.5μm), the mobile phase is 0.05mol / L phosphate buffer solution (pH6.8)-acetonitrile-methanol (17:7:6, V / V / V), the flow rate is 1.0ml / min, the detection wavelength is 230nm, the column temperature is 30℃, and the injection volume is 10μL. The content of active components and the salt formation rate of fumaric acid vonoprazan were studied. The systems used in the currently published literature are only suitable for the detection of other impurities in the raw materials or the content of the final product, and cannot effectively detect the above-mentioned pyridine-3-sulfonic acid and 4-dimethylaminopyridine. Therefore, it is necessary to develop a new detection method to effectively study the quality of the intermediate product 1 related substances produced in the synthesis process of fumaric acid vonoprazan. According to the existing background technology, pyridinesulfonic acid is a strongly acidic compound, and 4-dimethylaminopyridine is an alkaline compound, which is difficult to detect, retain and detect under the same conditions. Summary of the invention

[0011] Based on the HPLC conditions used in the above literature, it is impossible to detect the components to be measured.

[0012] In view of the above-mentioned defects of the prior art, the technical problem to be solved by the present invention is to develop a detection method that can simultaneously and accurately detect the related substances of vonoprazan fumarate intermediate 1, namely 4-dimethylaminopyridine and pyridine-3-sulfonic acid.

[0013] To achieve the above-mentioned purpose, the present invention provides a method for detecting related substances of vonoprazan fumarate intermediate 1, the method adopts high performance liquid chromatography, the chromatographic column filler adopts high-purity porous spherical silica gel particles coated with charged nano polymer microspheres, the detector is an ultraviolet detector, and the mobile phase system is an aqueous solution of ammonium acetate and acetonitrile.

[0014] Furthermore, its stationary phase uses nanohybrid silica polymer (NSH) technology, and the chromatographic column is recommended to use Acclaim TM Trinity TMP1 and P2 series chromatographic columns have a length of 100 mm to 250 mm and a particle size of 2.5 um to 5 um, preferably 3 um or 5 um.

[0015] Furthermore, the concentration of ammonium acetate in the mobile phase is 0.01M-0.05M, preferably 0.02M, and the pH range of ammonium acetate is 4.5-7.5, preferably 5.0.

[0016] Other conditions: the detection wavelength of the ultraviolet detector is 240nm~280nm, preferably 260nm; the flow rate is 0.4~1.2mL / min, preferably 0.5~0.8ml / min; the column temperature of the chromatographic column is 20~45℃, preferably 30℃; the chromatographic column is preferably Acclaim TM Trinity TM P1 column.

[0017] Furthermore, the above method specifically includes the following contents: Solution preparation: Blank solution: diluent (mobile phase); Control solution: Take appropriate amount of 4-dimethylaminopyridine and pyridine-3-sulfonic acid and dilute to 0.4 μg / ml; Test solution: weigh about 20 mg of intermediate product 1, place it in a 50 ml volumetric flask, add diluent to dissolve and dilute; Accurately measure 10 μL of the blank solution, reference solution, and test solution and inject them into the chromatograph. Calculate the impurity content according to the external standard method to complete the quantitative detection of the related substance 2 of the vonoprazan fumarate intermediate product 1.

[0018] The detection method of the invention is simple and fast to operate, and the prior art does not report a detection method for these two substances. The invention overcomes the defect that the two polar acid-base substances cannot be effectively retained in the same system according to the known prior art, and the retention time of each component is short. During use, the chromatographic system is durable and stable, and impurity residues can be effectively detected. The determination method has strong specificity, and the detection result is accurate and reliable. The invention provides a simple and reasonable detection method for residual detection of pyridine-3-sulfonic acid and 4-dimethylaminopyridine in the intermediate product 1 in the continuous batch production process.

[0019] The concept, specific structure and technical effects of the present invention will be further described below in conjunction with the accompanying drawings to fully understand the purpose, characteristics and effects of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Synthesis route of vonoprazan fumarate.

[0021] Figure 2 This is a specific test map of a preferred embodiment of the present invention. DETAILED DESCRIPTION

[0022] The following describes the preferred embodiments of the present invention according to the drawings in the specification to make the technical content clearer and easier to understand; the present invention can be embodied in many different forms of embodiments, and the protection scope of the present invention is not limited to the embodiments mentioned in the text. Example 1

[0023] Determination of Related Substances in Intermediate 1 of Vonoprazan Fumarate Testing conditions and instruments (1) Instrument conditions LC-2030C high performance liquid chromatograph (quaternary pump, degassing unit, UV detector, column oven, autosampler, system monitor, LBsolution) (Shimadzu Corporation, Japan); test sample (Chongqing Shenghuaxi Pharmaceutical Co., Ltd.), acetonitrile (honeywell chromatographic grade), other reagents were of analytical grade, and water was fresh ultrapure water.

[0024] According to the high performance liquid chromatography method (Chinese Pharmacopoeia 2015 General Principle 0512), using Acclaim TM Trinity TM P1 4.6mm×100mm, 3μm is the chromatographic column, the mobile phase is 0.02mol / L ammonium acetate buffer (glacial acetic acid adjusted to pH5.0): acetonitrile (4:6); the flow rate is 0.5ml per minute; the detection wavelength is 260nm, the column temperature is 30℃, and the injection volume is 10μl.

[0025] Diluent mobile phase.

[0026] Take an appropriate amount of the product for test solution, weigh it accurately, dissolve it with a diluent and quantitatively dilute it to make a solution containing about 0.4 mg per 1 ml.

[0027] Reference solution: Weigh appropriate amounts of 4-dimethylaminopyridine and pyridine-3-sulfonic acid reference substances, dissolve them in a diluent and quantitatively dilute them to make a solution containing approximately 0.4 μg of 4-dimethylaminopyridine and pyridine-3-sulfonic acid per 1 ml.

[0028] System suitability requires that in the chromatogram of the reference solution, 4-dimethylaminopyridine and pyridine-3-sulfonic acid elute in sequence, and the theoretical plate numbers of 4-dimethylaminopyridine and pyridine-3-sulfonic acid must not be less than 1000.

[0029] (2) Solution preparation: Blank solution: diluent (mobile phase) Reference substance solution (0.4μg / ml): Accurately weigh about 20mg each of 4-dimethylaminopyridine reference substance and pyridine-3-sulfonic acid reference substance, place them in a 50ml volumetric flask, add diluent to dissolve and dilute to the scale, shake well; accurately measure 1ml into a 100ml volumetric flask, dilute to the scale with diluent, shake well; accurately measure into a 10ml volumetric flask, dilute to the scale with diluent, shake well.

[0030] (3) Determination method: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram.

[0031] (4) If there is a chromatographic peak in the chromatogram of the test solution with the same retention time as that in the reference solution, calculate the impurity content using the impurity external standard method.

[0032] Validation of method specificity Solution preparation Blank solution: diluent (mobile phase).

[0033] Impurity stock solution: Accurately weigh about 10 mg of 4-dimethylaminopyridine reference substance and pyridine-3-sulfonic acid reference substance, respectively, and place them in 100 ml volumetric flasks, add diluent to dissolve and dilute to the scale, shake well, and prepare impurity stock solutions ① and ② respectively.

[0034] Preparation of impurity localization solution (10μg / ml): Take 1ml of each of the above impurity stock solutions ① and ②, place them in 10ml volumetric flasks, add diluent to dilute to the scale, shake well, and the localization solution of each impurity is obtained.

[0035] Mixed solution: Accurately weigh about 20 mg of intermediate product 1, place in a 50 ml volumetric flask, add 0.2 ml of the above impurity stock solutions ① and ② respectively, add diluent to dilute to the scale, shake well to obtain a mixed solution. (As shown in the figure Figure 2 shown) Accurately measure 10 μL of the blank, mixed solution, and each impurity location solution, inject them into the liquid chromatograph, and record the chromatogram.

[0036] Test results.

[0037] The intermediate product 1 peaks at 1.778min, the retention time of 4-dimethylaminopyridine is 3.577min, the separation degree from the front peak is 5.56, the separation degree from the rear peak is 14.77, and the peak of pyridine-3-sulfonic acid is 7.215min, the separation degree from the front peak is 14.77. The separation degree between the known impurities and the adjacent impurity peaks is greater than 1.5; and the blank solvent does not interfere with the detection of the analyte. The method has good specificity.

[0038] In addition to specificity, this method has also been validated by detection limit, linearity and range, solution stability, accuracy, repeatability, precision, and durability, demonstrating that this method can be stably used for the detection of related substances in vonoprazan fumarate intermediate 1, namely 4-dimethylaminopyridine and pyridine-3-sulfonic acid. Example 2

[0039] Four consecutive batches of intermediate products 1 produced by our company were tested and the test results are summarized as follows

[0040] It can be seen from the above table that in the intermediate product 1, relatively small amounts of 4-dimethylaminopyridine and pyridine-3-sulfonic acid can be detected.

[0041] The preferred specific embodiments of the present invention are described in detail above. It should be understood that ordinary technicians in the field can make many modifications and changes based on the concept of the present invention without creative work. Therefore, all technical solutions that can be obtained by technicians in the technical field based on the concept of the present invention through logical analysis, reasoning or limited experiments on the basis of the prior art should be within the scope of protection determined by the claims.

Claims

1. A method for detecting related substances in vonoprazan fumarate intermediates, characterized in that the detection method is carried out by high performance liquid chromatography, the chromatographic column filler is composed of high-purity porous spherical silica gel particles coated with charged nano polymer microspheres, the detector is an ultraviolet detector, and the mobile phase system is ammonium acetate aqueous solution and acetonitrile, and the related substances include: 4-Dimethylaminopyridine, pyridine-3-sulfonic acid.

2. The detection method according to claim 1, characterized in that: The chromatographic column filler adopts nano-hybrid silicon polymer technology.

3. The detection method according to claim 1, characterized in that The chromatographic column is Acclaim TM Trinity TM P1 or P2 series columns.

4. The detection method according to claim 1, characterized in that: The concentration of the ammonium acetate aqueous solution in the mobile phase is 0.01-0.05M.

5. The detection method according to claim 1, characterized in that: The pH range of the ammonium acetate aqueous solution in the mobile phase is 4.5-7.

5.

6. The detection method according to claim 1, characterized in that: The detection wavelength of the ultraviolet detector is 240nm~280nm.

7. The detection method according to claim 1, characterized in that: The flow rate of the mobile phase is 0.4-1.2 mL / min.

Images