A method for determining the content of compound I in olaparib sodium

Abstract

The application discloses a method for determining and analyzing the content of 2-fluoro-3-methoxyphenylboronic acid in olaparib sodium, which comprises the following steps: derivatizing the 2-fluoro-3-methoxyphenylboronic acid with N-methyliminodiacetic acid, determining the content of the 2-fluoro-3-methoxyphenylboronic acid derivative, and determining the content of the 2-fluoro-3-methoxyphenylboronic acid in the olaparib sodium. Compared with the prior art, the analysis method has the characteristics of good specificity, good reproducibility, extremely high sensitivity, and suitability for trace analysis.

Description

Technical Field

[0001] This invention relates to the determination of the content of 2-fluoro-3-methoxyphenylboronic acid derivative (hereinafter referred to as compound I derivative) in oxagogue sodium by using high performance liquid chromatography-mass spectrometry (HPLC-MS) to determine the content of 2-fluoro-3-methoxyphenylboronic acid derivative (hereinafter referred to as compound I derivative) after derivatization with N-methyliminodiacetic acid (hereinafter referred to as derivatizing agent). This method can effectively control the trace content of compound I in active pharmaceutical ingredients and formulations, providing assurance for the health and safety of patients' medication use, and belongs to the field of pharmaceutical technology. Technical Background

[0002] Oxalagolidin sodium, chemical name R-4-((2-(5-(2-fluoro-3-methoxyphenyl)-3-((2-fluoro-6-(trifluoromethyl)benzyl)-4-methyl-2,6-dioxo-2,3-dihydropyrimidin-1(6H)-yl)-1-phenylethyl)amino)butyrate sodium.

[0003] Elagolix sodium is an oral GnRH antagonist co-developed by AbbVie and its partner Neurocrine Biosciences. It works by inhibiting the pituitary gonadotropin-releasing hormone receptor, ultimately reducing circulating levels of sex hormones. Its structure is as follows:

[0004]

[0005] Oxalagolidin sodium contains compound I, a residue from the reaction, with the following structure:

[0006]

[0007] Accurately and quantitatively determining trace amounts of compound I in oxaragoline sodium is extremely challenging. Direct determination at trace levels using methods such as gas chromatography or liquid chromatography is difficult.

[0008] Currently, only methods for determining the content of macro-compound I, employing classical chromatography, suffer from significant deficiencies in specificity, sensitivity, method precision, and repeatability, failing to provide reliable trace analysis results. To overcome these shortcomings, this invention utilizes high-performance liquid chromatography-mass spectrometry (HPLC-MS / MS) to determine the content of compound I in oxagogueline sodium after derivatization with a derivatizing agent. This analytical method is characterized by its simplicity, high specificity, extremely high sensitivity, and excellent reproducibility. Summary of the Invention

[0009] As mentioned above, direct and accurate quantitative determination of trace compound I has various drawbacks. However, by utilizing the high reactivity of compound I with derivatizing agents, mild reaction conditions, the strong stability of its derivatives and their suitability for separation by reversed-phase liquid chromatography, as well as its easy ionization in a mass spectrometer detector and its stable molecular structure suitable for quantitative analysis, high-performance liquid chromatography-mass spectrometry (HPLC-MS / MS) can be used for determination. Furthermore, the sensitive response of compound I to the mass spectrometer detector greatly improves the sensitivity and specificity of this method.

[0010] The structure of the derivatizing agent N-methyliminodiacetic acid is as follows:

[0011]

[0012] The reaction route for the formation of derivative of compound I by reacting with the derivatizing agent N-methyliminodiacetic acid is as follows:

[0013] Experiments revealed that sodium oxaragoline exhibits excellent solubility in dimethyl sulfoxide (DMSO), and that compound I reacts efficiently with the derivatizing agent in acidic DMSO under mild and easily controlled conditions. This invention will use DMSO as a diluent.

[0014] This invention relates to an analytical method for determining the content of compound I in oxagogue sodium. The method involves reacting compound I with a derivatizing agent and determining the content of the derivative of compound I using high performance liquid chromatography-mass spectrometry, thereby indirectly determining the content of compound I in oxagogue sodium.

[0015] In the preparation of the above samples, a 10% acetic acid dimethyl sulfoxide solution was used as a diluent.

[0016] The above was performed using a high-performance liquid chromatography-mass spectrometry system in positive ion mode.

[0017] The method includes the following steps:

[0018] (1) Take sodium oxagogue raw material or preparation powder and use 5% to 30% dimethyl sulfoxide or N,N-dimethylformamide solution of formic acid or acetic acid or trifluoroacetic acid as diluent to prepare sample stock solution;

[0019] (2) Take the solid N-methyliminodiacetic acid derivative and use a 5% to 30% dimethyl sulfoxide or N,N-dimethylformamide solution of formic acid, acetic acid or trifluoroacetic acid as a diluent to prepare a derivatizing agent solution;

[0020] (3) Take the sample stock solution and the derivatizing agent solution and mix them evenly in a certain volume ratio. Heat them in an oven to 50 to 150°C for 20 minutes to 10 hours to prepare the sample solution.

[0021] (4) An octadecylsilane-bonded silica gel column was used, and the mobile phase was 0.05%-0.2% formic acid or acetic acid or trifluoroacetic acid aqueous solution and acetonitrile or methanol, and isocratic or gradient elution was performed.

[0022] (5) Set the mobile phase flow rate to 0.3-2.0 mL / min and control the column temperature between 25℃ and 45℃.

[0023] (6) Using a mass spectrometer detector, with the ion mode selected as positive ion, the sample solution from (3) was injected, and the mass ion chromatogram of the derivative of compound I was recorded.

[0024] The technical solution adopted in this invention is as follows:

[0025] Sample pretreatment:

[0026] Diluent: 5% to 30% dimethyl sulfoxide or N,N-dimethylformamide solution of formic acid, acetic acid, or trifluoroacetic acid.

[0027] Derivatizing agent solution: Weigh 100 mg of the derivatizing agent N-methyliminodiacetic acid into a 10 ml volumetric flask, dissolve and dilute to the mark with diluent, and shake well.

[0028] Compound I standard stock solution: Prepare a 200 ng / ml solution of Compound I using diluent.

[0029] Sample stock solution: Accurately weigh 400 mg of sample into a 10 ml volumetric flask, dissolve and dilute to the mark with diluent, and shake well.

[0030] Compound I standard solution: Mix 500 μl of compound I standard stock solution and 500 μl of derivatizing agent solution in a 1.5 ml vial and heat at 100 °C for 1 hour to obtain the standard solution.

[0031] Sample solution: Mix the sample stock solution with 500 μl of derivatizing agent solution in a 1.5 ml vial and heat at 100 °C for 1 hour to obtain the sample solution.

[0032] The chromatographic column used in this invention is an octadecylsilane-bonded silica column. The flow rate is 0.3-2.0 mL / min. The column temperature is 25℃-45℃. Mobile phase A is 0.05%-0.2% aqueous solution of formic acid, acetic acid, or trifluoroacetic acid, and mobile phase B is acetonitrile or methanol, with isocratic or gradient elution.

[0033] Mass spectrometer detector: positive ion mode, [M+H] + :282 Attached image description:

[0034] Appendix Figure 1 Mass spectrum bar graph of compound I derivative obtained according to Example 1 of the present invention;

[0035] Appendix Figure 2 The total ion chromatogram of compound I derivative obtained according to Example 2 of the present invention;

[0036] Appendix Figure 3 The total ion chromatogram of the sodium oxaragoline sample obtained according to Example 3 of the present invention;

[0037] Appendix Figure 4 The total ion chromatogram of the recovered sodium oxaragoline sample obtained according to Example 4 of the present invention; Detailed Implementation

[0038] To better understand the technical solution of the present invention, the following description is provided in conjunction with specific embodiments of the present invention, but these are not limited to the present invention.

[0039] Example 1

[0040] Instruments and conditions:

[0041] High performance liquid chromatography-mass spectrometry system: Agilent 1260 Infinity, MS detector.

[0042] Chromatographic column: Octadecylsilane-bonded silica gel column

[0043] Mobile phase: A: 0.05%-0.2% aqueous solution of formic acid, acetic acid, or trifluoroacetic acid; B: acetonitrile.

[0044] The isocratic conditions were as follows: 0–8.0 min, with an organic phase ratio of 5%.

[0045] Column temperature: 40℃.

[0046] Flow rate: 0.6 mL / min

[0047] Select positive ions: 282

[0048] Injection volume: 3ul.

[0049] Experimental steps:

[0050] 1) Preparation of mobile phase A: Accurately measure 1.0 mL of formic acid and dissolve it in 1000 mL of water, then mix well.

[0051] 2) Diluent: 10% acetic acid dimethyl sulfoxide solution

[0052] 3) Derivatizing agent solution: Weigh 100 mg of the derivatizing agent N-methyliminodiacetic acid into a 10 ml volumetric flask, dissolve and dilute to the mark with diluent, and shake well.

[0053] 4) Compound I Derivatization Solution: Accurately weigh approximately 20 mg of Compound I into a 10 ml volumetric flask, dissolve and dilute to the mark with diluent, and mix well to obtain the Compound I derivatization stock solution. Accurately transfer 0.5 ml of the Compound I derivatization stock solution into a 1.5 ml vial, then accurately transfer 0.5 ml of the derivatizing agent solution into the same vial and mix thoroughly. Heat in a 100°C oven for 1 hour, cool, and obtain the Compound I derivatization solution. Inject the sample and record the mass spectrum bar graph of the Compound I derivative. See typical... Figure 1 .

[0054] Example 2

[0055] Instruments and conditions:

[0056] High performance liquid chromatography-mass spectrometry system: Agilent 1260 Infinity, MS detector.

[0057] Chromatographic column: Octadecylsilane-bonded silica gel column

[0058] Mobile phase: A: 0.05%-0.2% aqueous solution of formic acid, acetic acid, or trifluoroacetic acid; B: acetonitrile.

[0059] The isocratic conditions were as follows: 0–8.0 min, with an organic phase ratio of 5%.

[0060] Column temperature: 40℃.

[0061] Flow rate: 0.6 mL / min

[0062] Select positive ions: 282

[0063] Injection volume: 3ul.

[0064] Experimental steps:

[0065] 1) Preparation of mobile phase A: Accurately measure 1.0 mL of formic acid and dissolve it in 1000 mL of water, then mix well.

[0066] 2) Diluent: 10% acetic acid in dimethyl sulfoxide solution.

[0067] 3) Derivatizing agent solution: Weigh 100 mg of the derivatizing agent N-methyliminodiacetic acid into a 10 ml volumetric flask, dissolve and dilute to the mark with diluent, and shake well.

[0068] 4) Compound I Standard Solution: Accurately weigh approximately 20 mg of Compound I into a 10 ml volumetric flask, dissolve and dilute to the mark with diluent, shake well. Accurately transfer 100 μl to another 10 ml volumetric flask, dilute to volume with diluent, shake well, and then accurately transfer another 100 μl to another 10 ml volumetric flask, shake well, and this gives the Compound I standard stock solution. Accurately transfer 0.5 ml of the Compound I standard stock solution into a 1.5 ml vial, and then accurately transfer 0.5 ml of the derivatizing agent solution into the same vial. Mix thoroughly, heat in a 100°C oven for 1 hour, cool, and this gives the Compound I standard solution. Inject the sample and record the total ion chromatogram of the Compound I derivative mass spectrum. See typical... Figure 2 .

[0069] Example 3

[0070] Instruments and conditions:

[0071] High performance liquid chromatography-mass spectrometry system: Agilent 1260 Infinity, MS detector.

[0072] Chromatographic column: Octadecylsilane-bonded silica gel column

[0073] Mobile phase: A: 0.05%-0.2% aqueous solution of formic acid, acetic acid, or trifluoroacetic acid; B: acetonitrile.

[0074] The isocratic conditions were as follows: 0–8.0 min, with an organic phase ratio of 5%.

[0075] Column temperature: 40℃.

[0076] Flow rate: 0.6 mL / min

[0077] Select positive ions: 282

[0078] Injection volume: 3ul.

[0079] Experimental steps:

[0080] 1) Preparation of mobile phase A: Accurately measure 1.0 mL of formic acid and dissolve it in 1000 mL of water, then mix well.

[0081] 2) Diluent: 10% acetic acid in dimethyl sulfoxide solution.

[0082] 3) Derivatizing agent solution: Weigh 100 mg of the derivatizing agent N-methyliminodiacetic acid into a 10 ml volumetric flask, dissolve and dilute to the mark with diluent, and shake well.

[0083] 4) Oxalgolin sodium sample solution: Accurately weigh approximately 400g of oxagolin sodium into a 10ml volumetric flask, dissolve and dilute to the mark with diluent, and shake well to obtain the oxagolin sodium sample stock solution. Accurately transfer 0.5ml of the oxagolin sodium sample stock solution into a 1.5ml vial, then accurately transfer 0.5ml of the derivatizing agent solution into the same vial and mix thoroughly. Heat in a 100℃ oven for 1 hour, cool, and obtain the oxagolin sodium sample solution. Inject the sample and record the total ion chromatogram of the oxagolin sodium sample mass spectrometry. See typical... Figure 3 .

[0084] Example 4

[0085] Instruments and conditions:

[0086] High performance liquid chromatography-mass spectrometry system: Agilent 1260 Infinity, MS detector.

[0087] Chromatographic column: Octadecylsilane-bonded silica gel column

[0088] Mobile phase: A: 0.05%-0.2% aqueous solution of formic acid, acetic acid, or trifluoroacetic acid; B: acetonitrile.

[0089] The isocratic conditions were as follows: 0–8.0 min, with an organic phase ratio of 5%.

[0090] Column temperature: 40℃.

[0091] Flow rate: 0.6 mL / min

[0092] Select positive ions: 282

[0093] Injection volume: 3ul.

[0094] Experimental steps:

[0095] 1) Preparation of mobile phase A: Accurately measure 1.0 mL of formic acid and dissolve it in 1000 mL of water, then mix well.

[0096] 2) Diluent: 10% acetic acid in dimethyl sulfoxide solution.

[0097] 3) Derivatizing agent solution: Weigh 100 mg of the derivatizing agent N-methyliminodiacetic acid into a 10 ml volumetric flask, dissolve and dilute to the mark with diluent, and shake well.

[0098] 4) Compound I standard stock solution: Accurately weigh about 20 mg of compound I into a 10 ml volumetric flask, add diluent to dissolve and dilute to the mark, shake well, accurately transfer 100 μl into another 10 ml volumetric flask, add diluent to make up to the mark, shake well, then accurately transfer another 100 μl into another 10 ml volumetric flask, shake well, and the compound I standard stock solution is obtained.

[0099] 5) Oxalgolin sodium sample recovery solution: Accurately weigh approximately 400 mg of oxagolin sodium into a 10 mL volumetric flask, dissolve and dilute to volume with Compound I standard stock solution, and shake well to obtain the oxagolin sodium sample recovery stock solution. Accurately transfer 0.5 mL of the oxagolin sodium sample recovery stock solution into a 1.5 mL vial, then accurately transfer 0.5 mL of the derivatizing agent solution into the same vial and mix thoroughly. Heat in a 100 °C oven for 1 hour, cool, and obtain the oxagolin sodium sample recovery solution. Inject the sample and record the total ion chromatogram of the oxagolin sodium sample recovery mass spectrometry. See typical... Figure 4 .

Claims

1. A method for determining the content of genotoxic impurity compound I in oxaragoline sodium, characterized in that, The compound I is 2-fluoro-3-methoxyphenylboronic acid. The content of trace compound I in oxaragoline sodium was determined by high performance liquid chromatography-mass spectrometry using a derivatization method. The method includes the following steps: (1) Take sodium oxagoguel raw material or preparation powder and use 5% to 30% dimethyl sulfoxide or N,N-dimethylformamide solution of formic acid or acetic acid or trifluoroacetic acid as diluent to prepare sample stock solution; (2) Take solid N-methyliminodiacetic acid as the derivatizing agent and use 5% to 30% dimethyl sulfoxide or N,N-dimethylformamide solution of formic acid, acetic acid or trifluoroacetic acid as diluent to prepare a derivatizing agent solution; (3) Take the sample stock solution and the derivatizing agent solution and mix them evenly in a certain volume ratio. Heat them in an oven to 50 to 150°C for 20 minutes to 10 hours to prepare the sample solution. (4) An octadecylsilane-bonded silica gel column was used, and the mobile phase was 0.05%-0.2% formic acid or acetic acid or trifluoroacetic acid aqueous solution and acetonitrile or methanol, with isocratic or gradient elution. (5) Set the mobile phase flow rate to 0.3-2.0 mL / min and control the column temperature between 25℃ and 45℃; (6) A mass spectrometer detector was used, with the ion mode selected as positive ion, [M+H]. + :282, take the sample solution of (3) and inject it, and record the mass ion chromatogram of compound I derivative.

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