Dexrazoxane impurity b and process for its preparation and use

Abstract

The application provides dextrorotatory razoxane impurity B, a preparation method and application thereof, and belongs to the technical field of drug synthesis. The structural formula of the dextrorotatory razoxane impurity B provided by the application is shown as formula I. The impurity B is a newly found impurity, the structure confirmation and synthesis of the impurity B solve the structure problem of impurities in the synthesis process of dextrorotatory razoxane, and provide sufficient theoretical support and research material basis for impurity research of the bulk drug and quality standard establishment of the bulk drug.

Description

Technical Field

[0001] This invention relates to the field of drug synthesis technology, and in particular to dexrazoxane impurity B, its preparation method, and its application. Background Technology

[0002] Dexrazoxane is an adjuvant antitumor drug clinically used to alleviate or reduce cardiotoxicity caused by anthracycline antibiotics (such as doxorubicin) chemotherapy. The mechanism by which dexrazoxane exerts its cytoprotective activity is not fully understood. Dexrazoxane is a lipophilic derivative of the chelating agent ethylenediaminetetraacetic acid (EDTA), which can rapidly cross the cell membrane to reduce the cardiotoxicity of anthracycline antitumor antibiotics such as doxorubicin. Intracellularly, dexrazoxane hydrolyzes to TCRE-198, which then chelates with intracellular iron, reducing the complex of ferric ions with anthracycline antitumor drugs such as doxorubicin and preventing the formation of free radicals, thus exerting its effect.

[0003] The chemical name of drezosen is (S)-4,4'-(1-methyl-1,2-bimethylene)-bis(2,6-piperazinedione), and its structure is shown below:

[0004]

[0005] As the main active ingredient in pharmaceuticals, the impurities in active pharmaceutical ingredients (APIs) have always been a key research focus. Impurities exceeding the limits can alter physicochemical constants, affecting drug stability and efficacy, leading to lower drug content or reduced activity, increased toxicity and side effects, and even adverse effects on human health, resulting in immeasurable consequences. Therefore, impurity research in pharmaceuticals is a crucial step in controlling drug purity and improving drug quality. It also provides a basis for ensuring drug safety and efficacy, quality assurance during production and distribution, and performance evaluation of enterprise management.

[0006] Given the special nature of pharmaceuticals as commodities, their safety and efficacy are closely related to human life and health. Identifying the hazards of drug impurities and determining their toxicity during the production process is a crucial aspect of drug development. This research spans the entire process of drug development, production, and sales. To meet the technical requirements for active pharmaceutical ingredient (API) applications, advance clinical research to improve drug safety, and further control drug product quality, impurity research is indispensable. Summary of the Invention

[0007] In view of this, the present invention provides impurity B of dexrazoxane, its preparation method, and its application. The present invention provides a reference standard for the quantitative and qualitative analysis of dexrazoxane raw materials and / or preparations, thereby improving the quality standards of dexrazoxane and enhancing its quality and safety.

[0008] The technical solution of the present invention is as follows:

[0009] This invention provides a dextrozosin B impurity, the structural formula of which is shown in Formula I:

[0010]

[0011] This invention provides a method for preparing dextrozoxin impurity B, comprising the following steps:

[0012] (1) Mix 1,2-propanediamine, tert-butyl chloroacetate, organic solvent and alkaline reagent, react at -5 to 5°C for 0.5 to 13 hours, then react at room temperature for 8 to 40 hours, remove the solvent by vacuum distillation to obtain an oily substance;

[0013] (2) The oily substance prepared in step (1) is mixed with concentrated hydrochloric acid and organic solvent and reacted at 20-100℃. After the reaction is completed, the reaction solution is evaporated to dryness under reduced pressure at 40-72℃, dissolved in water, and then separated using a chromatographic column to obtain impurity B.

[0014] In this invention, the reaction equation for the above preparation method is as follows:

[0015]

[0016] This invention provides a method for preparing dextrozoxin impurity B, comprising the following steps:

[0017] S1. Mix 1,2-propanediamine, tert-butyl chloroacetate, organic solvent, and alkaline reagent. First, react at -5 to 5°C for 0.5 to 13 hours, then react at room temperature for 8 to 40 hours. Remove the solvent by vacuum distillation to obtain an oily substance.

[0018] S2. Mix the oily substance prepared in step S1, Boc anhydride, organic solvent and alkaline reagent, stir and react at room temperature for 2-48 hours, then add water and dichloromethane to the reaction solution for extraction, combine the organic phases, dry, filter, remove the solvent under reduced pressure to obtain the residue.

[0019] S3. The residue obtained in step S2 is mixed with acid and organic solvent and stirred at room temperature. After the reaction is complete, the solvent is removed by vacuum distillation, dissolved in water, and then separated using a chromatographic column to obtain impurity B.

[0020] In this invention, the reaction equation for the above preparation method is as follows:

[0021]

[0022] In one specific embodiment, the organic solvent is at least one selected from methanol, ethanol, dichloromethane, tetrahydrofuran, acetone, isopropanol, dioxane, and ethyl acetate;

[0023] The alkaline reagent is at least one selected from triethylamine, N,N-diisopropylethylamine, potassium carbonate, cesium carbonate, potassium hydroxide, and sodium hydroxide.

[0024] The acid is trifluoroacetic acid and / or hydrochloric acid.

[0025] In this invention, any alkaline reagent that can provide an alkaline environment is acceptable.

[0026] In one specific embodiment, the chromatographic column is an AQC18 aqueous column, and the preparative chromatographic conditions are as follows:

[0027] Filler: Octadecylsilane-bonded silica gel filler

[0028] Detection wavelength: 200-214nm

[0029] Flow rate: 15-40 ml / min

[0030] Injection volume: 2-4 ml

[0031] Mobile phase: Methanol or acetonitrile and 0.1-0.3% formic acid aqueous solution eluted at a ratio of 0-10%:100-90%.

[0032] In this invention, the chromatographic column separation and purification of impurity B is isocratic elution.

[0033] In one specific embodiment, in step (1), the mass ratio of 1,2-propanediamine to tert-butyl chloroacetate is 2:4.5-13.4; in step (2), the ratio of the oily substance to concentrated hydrochloric acid is 3.6g:3.6-36ml; and the mass fraction of concentrated hydrochloric acid is 37%.

[0034] In one specific embodiment, in step S1, the mass ratio of 1,2-propanediamine to tert-butyl chloroacetate is 2:4.5-13.4; in step S2, the ratio of oil to Boc anhydride is 3.9g:4.5-8.8g; and in step S3, the ratio of residue to acid is 1g:0.1-3g.

[0035] This invention also provides the application of dexrazoxane impurity B as an impurity reference standard in the quality control of dexrazoxane and its salts in the drug substance and its preparations.

[0036] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0037] 1. Impurity B of this invention is a newly discovered impurity. The structural confirmation and synthesis of the two impurities solve the structural problems of impurities in the synthesis of dexrazoxane, providing sufficient theoretical support and material basis for the research on impurities in active pharmaceutical ingredients and the formulation of quality standards for active pharmaceutical ingredients.

[0038] 2. The formation mechanism of impurity B is that it is obtained by intramolecular condensation and ring synthesis under acidic conditions of the intermediate. This impurity is a novel compound. It provides experimental data for the theoretical derivation of impurity formation in the process development of dextromethorphan active pharmaceutical ingredient (API), provides optimized basic data for further improving the quality of dextromethorphan API, promotes its clinical research, improves drug safety, and is of great significance for further controlling the quality of drug products. Attached Figure Description

[0039] Figure 1 The structure diagram of impurity B in Example 1 is obtained by 1H NMR spectroscopy.

[0040] Figure 2 This is a mass spectrometry analysis result of impurity B in Example 1;

[0041] Figure 3 This is the HPLC purity chromatogram of impurity B in Example 1;

[0042] Figure 4 The image shows the HPLC chromatogram of dexrazoxane in Example 4. Detailed Implementation

[0043] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0044] Example 1: Preparation of dextrozosen impurity B

[0045] Substitution reaction: 1,2-propanediamine (2.0 g), tert-butyl chloroacetate (4.5 g), 20 mL of dichloromethane and triethylamine (9.6 g) were added to a reaction flask. The reaction was first carried out at 5 °C for 13 hours, and then at room temperature for 40 hours. The solvent was removed by vacuum distillation to obtain 7.77 g of oil.

[0046] Cyclic reaction: Add the above oily substance (3.6g), 10ml of 37% con.HCl, and 10mL of THF to the reaction flask. Stir the reaction at 0℃ and monitor by thin-layer chromatography (TLC). After the reaction is complete, evaporate the reaction solution to dryness under reduced pressure at 72℃. Dissolve the residue in pure water and separate and purify using an AQC18 column.

[0047] Filler: Octadecylsilane-bonded silica gel filler

[0048] Detection wavelength: 200-214nm

[0049] Flow rate: 15-40 ml / min

[0050] Injection volume: 2-4 ml

[0051] Mobile phase: Methanol or acetonitrile and 0.1-0.3% formic acid aqueous solution eluted at a ratio of 0-10%:100-90%.

[0052] The elution time of impurity B was 10-20 minutes, and the yield of impurity B was 425 mg (17.9%).

[0053] The structure of impurity B is confirmed as follows:

[0054] Structure determined by proton nuclear magnetic resonance spectroscopy as follows Figure 1 As shown.

[0055] Mass spectrometry analysis results are shown below Figure 2 As shown, the molecular weight is 230.

[0056] The structure of impurity B is as follows:

[0057]

[0058] Purity identification of impurity B: Impurity B was detected by HPLC under the following conditions:

[0059] Column specifications: C18 (filler: octadecylsilane-bonded silica gel filler)

[0060] Column temperature: 30℃

[0061] Flow rate: 1.0 ml / min

[0062] Detection wavelength: 214nm

[0063] Injection volume: 2uL

[0064] Mobile phase A: Methanol (100%) Mobile phase B: 0.03% phosphoric acid

[0065] Gradient elution procedure:

[0066] 0 5％ 95％ 30 95％ 5％

[0067] The chromatogram of impurity B as determined by HPLC is shown below. Figure 3 As shown, the retention time of impurity B is 4.147 min, and the purity is 91.977%.

[0068] Example 2: Preparation of dextrozosen impurity B

[0069] Substitution reaction: 1,2-Propanediamine (2.0 g), tert-butyl chloroacetate (13.4 g), 20 mL of dichloromethane and triethylamine (9.6 g) were added. The reaction was carried out at 5 °C for 13 hours, and then at room temperature for 40 hours. The solvent was removed by vacuum evaporation to obtain 7.77 g of oil.

[0070] Cycling reaction: Add 3.6 g of oil, 10 mL of 37% con.HCl, and 10 mL of acetonitrile to the reaction flask. Stir the reaction at 0 °C and monitor by thin-layer chromatography (TLC). After the reaction is complete, evaporate the reaction solution to dryness under reduced pressure at 72 °C. The residue is separated and purified in the same way as in Example 1. Impurity B 460 mg (yield 20.1%).

[0071] Impurity B was structurally confirmed to be consistent with that of Example 1.

[0072] Example 3: Preparation of dextrozosen impurity B

[0073] Substitution reaction: 1,2-Propanediamine (1.0 g), tert-butyl chloroacetate (6.2 g), 20 mL of dichloromethane and triethylamine (4.8 g) were added. The reaction was carried out at 5 °C for 13 hours, and then at room temperature for 40 hours. The solvent was removed by vacuum distillation to obtain 3.9 g of oil.

[0074] Boc protection: The prepared oil (3.9 g), Boc anhydride (4.5 g), 30 mL of ethanol:water = 2:1 and potassium carbonate (12.3 g) were added to the reaction flask. The mixture was stirred at room temperature for 2 h. Thin-layer chromatography (TLC) was used to monitor the reaction. After the reaction was completed, water and dichloromethane were added to the reaction solution for extraction. The organic phases were combined, dried, filtered, and the solvent was removed by vacuum evaporation to obtain 5.5 g of residue.

[0075] Cyclic reaction: Add the above-prepared residue (2.5 g), trifluoroacetic acid (2 g), and 10 mL of dichloromethane to the reaction flask. Stir the reaction at room temperature and monitor by thin-layer chromatography (TLC). After the reaction is complete, remove the solvent under reduced pressure to obtain an oily liquid. Follow the same separation and purification steps as in Example 1. Impurity B 360 mg (yield 15.7%).

[0076] Impurity B was structurally confirmed to be consistent with that of Example 1.

[0077] Example 4

[0078] Detection of impurity B in dextromethorphan:

[0079] Source of dexrazoxane: Commercially available dexrazoxane.

[0080] The levozosen was analyzed by HPLC under the same conditions as the HPLC purity test conditions for impurity B in Example 1.

[0081] The HPLC chromatogram of levozosin is shown below. Figure 4 As shown, the peak with a retention time of 4.151 min is impurity B. After collecting impurity B, it was confirmed again by structural confirmation. The structural confirmation result was consistent with that of Example 1, thus determining the presence of impurity B in dexrazoxane.

[0082] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing dextrozosen impurity B, wherein the structural formula of dextrozosen impurity B is shown in Formula I: Its characteristics are, Includes the following steps: (1) Mix 1,2-propanediamine, tert-butyl chloroacetate, organic solvent, and triethylamine, and react them at -5 to 5°C for 0.5 to 13 hours, then at room temperature for 8 to 40 hours. Remove the solvent by vacuum distillation to obtain an oily substance; the mass ratio of 1,2-propanediamine to tert-butyl chloroacetate is 2:4.5-13.

4. (2) The oily substance prepared in step (1) is mixed with concentrated hydrochloric acid and organic solvent and reacted at 20-100℃. After the reaction is completed, the reaction solution is evaporated to dryness under reduced pressure at 40-72℃, dissolved in water, and then separated by chromatographic column to obtain impurity B; the ratio of the amount of oily substance to concentrated hydrochloric acid is 3.6g:3.6-36ml. The chromatographic column used was an AQC18 aqueous column, and the preparative chromatographic conditions were as follows: Filler: Octadecylsilane-bonded silica gel filler; Detection wavelength: 200-214nm; Flow rate: 15-40 ml / min; Injection volume: 2-4 ml; Mobile phase: Methanol or acetonitrile and 0.1-0.3% formic acid aqueous solution eluted at a ratio of 0-10%:100-90%; The peak time is 10-20 minutes.

2. A method for preparing dextrozosen impurity B, wherein the structural formula of dextrozosen impurity B is shown in Formula I: Its characteristics are, Includes the following steps: S1. Mix 1,2-propanediamine, tert-butyl chloroacetate, organic solvent, and triethylamine, and react at -5 to 5°C for 0.5 to 13 hours, then react at room temperature for 8 to 40 hours. Remove the solvent under reduced pressure to obtain an oily substance; the mass ratio of 1,2-propanediamine to tert-butyl chloroacetate is 2:4.5-13.

4. S2. Mix the oily substance prepared in step S1, Boc anhydride, organic solvent and potassium carbonate, stir and react at room temperature for 2-48 hours, then add water and dichloromethane to the reaction solution for extraction, combine the organic phases, dry, filter, and remove the solvent under reduced pressure to obtain the residue; the ratio of the oily substance to Boc anhydride is 3.9g:4.5-8.8g. S3. The residue obtained in step S2 is mixed with trifluoroacetic acid and an organic solvent, and stirred at room temperature. After the reaction is complete, the solvent is removed by vacuum distillation, the residue is dissolved in water, and then separated by chromatographic column to obtain impurity B. The ratio of the residue to acid is 1g:0.1-3g. The chromatographic column used was an AQC18 aqueous column, and the preparative chromatographic conditions were as follows: Filler: Octadecylsilane-bonded silica gel filler; Detection wavelength: 200-214nm; Flow rate: 15-40 ml / min; Injection volume: 2-4 ml; Mobile phase: Methanol or acetonitrile and 0.1-0.3% formic acid aqueous solution eluted at a ratio of 0-10%:100-90%; The peak time is 10-20 minutes.

3. The method for preparing dexrazoxan impurity B according to claim 1 or 2, characterized in that, The organic solvent is at least one of methanol, ethanol, dichloromethane, tetrahydrofuran, acetone, isopropanol, dioxane, and ethyl acetate.

4. The method for preparing dextrozoxin impurity B according to claim 1, characterized in that, In step (2), the mass fraction of concentrated hydrochloric acid is 37%.

5. Application of dexrazoxane impurity B as an impurity reference standard in the quality control of dexrazoxane and its salts in active pharmaceutical ingredients and preparations; the structural formula of dexrazoxane impurity B is shown in Formula I: .

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