Epristeride impurity 17 beta-(N-tertiary butyl-amino-formyl) androstane-3, 5-diene-3-diphenylphosphine oxide as well as preparation method and application of epristeride impurity 17 beta-(N-tertiary butyl-amino-formyl) androstane-3, 5-diene-3-diphenylphosphine oxide

Abstract

The invention discloses an epristeride impurity 17 beta-(N-tertiary butyl-amino-formyl) androstane-3, 5-diene-3-diphenylphosphine oxide as well as a preparation method and application thereof, and belongs to the field of medical chemistry. Comprising the following steps: 1) taking an epristeride grignard substance as a raw material, and carrying out a halide catalytic reaction on the epristeride grignard substance and a phosphine reagent in an organic solvent under an alkaline condition to obtain an epristeride impurity crude product reaction solution; and 2) performing acidolysis treatment on the epristeride impurity crude product reaction solution, concentrating to obtain an oily substance, and performing column chromatography purification on the oily substance to obtain the epristeride impurity. The method is simple and efficient to operate, mild in reaction condition, easy to control and high in safety, the epristeride impurity prepared by the method is high in yield, the purity is as high as 97.6%, the epristeride impurity is suitable for being used as a reference substance to research the quality of epristeride bulk drugs and preparations thereof, and a foundation is laid for effective control of the quality of epristeride related drugs.

Description

Technical Field

[0001] The invention belongs to the field of pharmaceutical chemistry, and specifically relates to an aprelet impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, and a preparation method and application thereof. Background Art

[0002] Epristeride, also known as epristamide, has a chemical name of 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-carboxylic acid. This product can form a ternary complex with 5α-reductase and nicotinamide adenine dinucleotide phosphate, thereby inhibiting the conversion of testosterone into dihydrotestosterone and reducing the content of dihydrotestosterone in the prostate, leading to the atrophy of the hyperplastic prostate, and achieving the purpose of inhibiting prostate hyperplasia.

[0003] In order to ensure the safety of the drug use of Aprelet and to achieve the purpose of effectively controlling its quality, the study of impurities in drugs is particularly important. At present, the internationally recognized method is to identify impurities by analyzing impurity reference substances, so it is of great significance to carry out the synthesis research of impurities in Aprelet. Compound I is a common impurity in the synthesis of Aprelet. After searching, there is currently no method for synthesizing this impurity, so the impurity is synthesized to confirm its structure, study its physical and chemical properties so as to control it, and select a suitable method to effectively remove this impurity, which is of great significance to improve the quality of Aprelet. Summary of the invention

[0004] The technical problem solved by the present invention is to provide an aplite impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide; another technical problem to be solved by the present invention is to provide a preparation method of the aplite impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide; another technical problem to be solved by the present invention is to provide an application of the aplite impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide as a reference substance for quality research of aplite raw materials and preparations thereof.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0006] An impurity of Aprelet, 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, has a structural formula as shown in Formula I:

[0007]

[0008] The preparation method of the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet comprises the following steps:

[0009] 1) using 7β-(N-tert-butyl-amino-formyl)androsta-3-bromo-3,5-diene as a raw material, reacting with a phosphine reagent under alkaline conditions in an organic solvent, and obtaining a crude reaction solution of 7β-(N-tert-butyl-amino-formyl)androsta-3-bromo-3,5-diene as a raw material ... reacting with a phosphine reagent under alkaline conditions in an organic solvent, and under halide catalysis;

[0010] 2) The crude reaction solution of Aprelete impurity is concentrated by post-acid hydrolysis to obtain an oily substance, and the oily substance is purified by column chromatography to obtain Aprelete impurity;

[0011] The reaction formula is as follows:

[0012]

[0013] In the step 1), the mass ratio of the aplite Grignard compound, the phosphine reagent, the alkali reagent and the catalyst is 1:0.4-0.6:1.3-1.5:0.2-0.6.

[0014] In the step 1), the mass volume ratio of the aplite Grignard compound to the organic solvent is 1:5-10.

[0015] The phosphine reagent in step 1) is diphenylphosphine oxide or triphenylphosphine.

[0016] The alkaline conditions in step 1) include at least one of dimethylethylenediamine, 1,8-diazabicycloundec-7-ene, triethylamine, cesium carbonate, sodium carbonate, potassium carbonate, inorganic base or organic base, or a mixture of two thereof.

[0017] The catalyst in step 1) is at least one of cuprous iodide and sodium iodide, or a mixture of the two.

[0018] The organic solvent in step 1) includes at least one of toluene, tetrahydrofuran, dimethylformamide and DMSO.

[0019] The reaction temperature of step 1) is 100-110° C., and the reaction time is 20-48 hours.

[0020] The above-mentioned aplite impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide is used as a reference substance in the quality research of aplite raw materials and preparations.

[0021] Compared with the prior art, the present invention has the following advantages:

[0022] The present invention provides for the first time a method for preparing an eplast impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, which has simple and efficient operation, mild reaction conditions, easy control and strong safety. The eplast impurity prepared by the method has a high yield and a purity of up to 97.6%, and is suitable for being used as a reference substance to control the quality of an eplast raw material and its preparations, thus laying a solid foundation for effectively controlling the quality of eplast-related drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet prepared by the method of Example 1 of the present invention is 1 H-NMR spectrum;

[0024] Figure 2 The impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet prepared by the method of Example 1 of the present invention is 13 C-NMR spectrum;

[0025] Figure 3 This is the mass spectrum of the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet prepared by the method of Example 1 of the present invention;

[0026] Figure 4 This is the HPLC spectrum of the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet prepared according to the method of Example 1 of the present invention. DETAILED DESCRIPTION

[0027] The present invention is further illustrated below in conjunction with specific examples. The examples are implemented based on the technical solutions of the present invention. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0028] Example 1

[0029] A method for preparing an impurity of Aprelet, 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, comprises the following steps:

[0030] (1) Weigh 10 g of eprite Grignard, add 4.23 g of diphenylphosphine oxide, 0.55 g of dimethylethylenediamine, 13.64 g of cesium carbonate, 0.4 g of cuprous iodide, 4.7 g of sodium iodide, and 100 mL of toluene into a reaction flask, replace with nitrogen three times, stir and heat to 100-110° C. for 20 h, and obtain a crude reaction solution of eprite impurities after the reaction is completed.

[0031] (2) The reaction solution of the crude product of Aprelet impurity obtained in step (1) was cooled to room temperature, the insoluble matter was removed by filtration, 2 mol / L hydrochloric acid was added to the organic phase, stirred and reacted for 2 h, the water layer was separated, the organic phase was washed with water and concentrated to dryness; 2.8 g of light yellow solid was obtained by column chromatography, namely Aprelet impurity, with a mass yield of 28.0% and a purity of 97.6%.

[0032] like Figure 1 As shown, the nuclear magnetic resonance hydrogen spectrum of the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide obtained in Example 1 ( 1 H-NMR instrument: BRUKER AV-400 nuclear magnetic resonance instrument) analysis shows that in CDCl3 as solvent 1 H-NMR, excluding the solvent peak of CDCl3, measured the resonance peaks of 46 protons in 11 different environments in the molecular structure, and the integral ratio of each peak (from low field to high field) was: 4:3:3:13:3:5:1:1:1:1:10. The type and number of protons in the test sample were consistent with the characteristics of hydrogen atoms in the molecular structure of the impurity of Aprelet.

[0033] like Figure 2 As shown, the carbon spectrum of the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide obtained in Example 1 ( 13 C-NMR instrument: BRUKER AV-400 nuclear magnetic resonance instrument) analysis shows that in CDCl3 as solvent 13 In the C-NMR spectrum, apart from the resonance peak of CDCl3 (δC77.38, 77.27, 77.06, 76.75 in the figure), there are 36 resonance peaks of carbon in 26 different environments with chemical shifts from large to small. The type and number of carbon nuclei in the test sample are consistent with the carbon atom characteristics in the impurity molecular structure.

[0034] like Figure 3 As shown, the liquid phase mass spectrum (LC-MS instrument: agilent6120 LC-MS liquid chromatography mass spectrometer) of the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide obtained in Example 1 shows that its molecular weight is 556.3. The results are consistent with the structural characteristics of the impurity of Aprelet.

[0035] Example 2

[0036] A method for preparing an impurity of Aprelet, 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, comprises the following steps:

[0037] (1) Weigh 10 g of Aplite Grignard, add 5.95 g of triphenylphosphine, 0.95 g of potassium carbonate, 13.64 g of cesium carbonate, 0.4 g of cuprous iodide, 4.7 g of sodium iodide, and 100 mL of toluene into a reaction flask, replace with nitrogen three times, stir and heat to 100-110° C. for 24 h, and obtain a crude Aplite impurity reaction solution after the reaction is completed.

[0038] (2) The reaction solution of the crude product of Aprelet impurity obtained in step (1) was cooled to room temperature, the insoluble matter was removed by filtration, 2 mol / L hydrochloric acid was added to the organic phase, stirred and reacted for 2 h, the water layer was separated, the organic phase was washed with water and concentrated to dryness; 2.1 g of light yellow solid was obtained by column chromatography, namely Aprelet impurity, with a mass yield of 21.0% and a purity of 95.8%.

[0039] Example 3

[0040] A method for preparing an impurity of Aprelet, 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, comprises the following steps:

[0041] (1) Weigh 10 g of eprite Grignard, add 5.95 g of triphenylphosphine, 0.95 g of potassium carbonate, 12.74 g of 1,8-diazabicycloundec-7-ene, 1.2 g of cuprous iodide, 0.47 g of sodium iodide, and 100 mL of tetrahydrofuran into a reaction flask, replace the atmosphere with nitrogen three times, stir and heat to 100-110° C., react for 48 h, and obtain a crude eprite impurity reaction solution after the reaction is completed.

[0042] (2) The reaction solution of the crude product of Aprelet impurity obtained in step (1) was cooled to room temperature, the insoluble matter was removed by filtration, the organic phase was concentrated to dryness and then dissolved in 100 mL of ethyl acetate, and stirred with 2 mol / L hydrochloric acid for 2 h, the water layer was separated, and the organic phase was washed with water and concentrated to dryness; 1.8 g of light yellow solid was obtained by column chromatography, namely, Aprelet impurity was obtained, with a mass yield of 18.0% and a purity of 96.1%.

[0043] Example 4

[0044] A method for preparing an impurity of Aprelet, 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, comprises the following steps:

[0045] (1) Weigh 10 g of Aplite Grignard, add 4.23 g of diphenylphosphine oxide, 0.95 g of potassium carbonate, 12.74 g of 1,8-diazabicycloundec-7-ene, 2.4 g of cuprous iodide, and 100 mL of DMSO into a reaction bottle, replace the atmosphere with nitrogen three times, stir and heat to 100-110° C., react for 48 hours, and obtain a crude Aplite impurity reaction solution after the reaction is completed.

[0046] (2) The reaction solution of the crude product of Aprelet impurity obtained in step (1) was cooled to room temperature, the insoluble matter was removed by filtration, the organic phase was concentrated to dryness and then dissolved in 100 mL of ethyl acetate, and stirred with 2 mol / L hydrochloric acid for 2 h, the water layer was separated, and the organic phase was washed with water and concentrated to dryness; 1.3 g of light yellow solid was obtained by column chromatography, namely, Aprelet impurity was obtained, with a mass yield of 13.0% and a purity of 96.5%.

[0047] Example 5

[0048] A method for preparing an impurity of Aprelet, 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, comprises the following steps:

[0049] (1) Weigh 10 g of Aplite Grignard, add 4.23 g of diphenylphosphine oxide, 0.95 g of potassium carbonate, 12.74 g of 1,8-diazabicycloundec-7-ene, 2.4 g of cuprous iodide, and 50 mL of dimethylformamide into a reaction flask, replace the atmosphere with nitrogen three times, heat to 100-110° C. with stirring, and react for 48 hours. After the reaction is completed, a crude Aplite impurity reaction solution is obtained.

[0050] (2) The reaction solution of the crude product of Aprelet impurity obtained in step (1) was cooled to room temperature, the insoluble matter was filtered out, 500 mL of water was added to the organic phase and stirred to dissolve, 2 mol / L hydrochloric acid was added and stirred to react for 2 h, and 200 mL of ethyl acetate was extracted. The organic phase was washed with water and concentrated to dryness; 1.2 g of yellow solid was obtained by column chromatography, i.e., Aprelet impurity was obtained, with a mass yield of 12.0% and a purity of 95.1%.

[0051] Example 6

[0052] Application of the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide in the quality study of Aprelet drug substance and its preparations

[0053] HPLC: Liquid chromatograph: Shimadzu LC-20AT; Chromatographic column: Inert Sustain C18 column (4.6×250mm, 5μm). Dissolve 25mL of glacial acetic acid and 1mL of diethylamine in water and dilute to 1000mL, mix well as mobile phase A, use methanol as mobile phase B, detection wavelength is 260nm, flow rate is 1.0mL / min, column temperature is 35℃. The injection volume is 10μL. Elution is performed according to the following gradient program:

[0054] Time (min) Mobile phase A (%) Mobile phase B (%) 0 30 70 30 30 70 30.1 17 83 50 17 83

[0055] Take appropriate amount of Aprelet and Aprelet impurities, weigh them, dissolve them with ultrasonic diluent, and prepare a mixed solution containing about 2.5 mg of Aprelet and 2.5 μg of Aprelet impurities per 1 mL as the system suitability solution. Perform HPLC analysis under the above conditions and record the chromatogram, such as Figure 4 As shown, the elution order is Apelite, Apelite impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide and other impurities. Under the above chromatographic conditions, the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Apelite is well separated from Apelite, and the theoretical plate number of each impurity is not less than 4000, which meets the requirements of the Chinese Pharmacopoeia.

[0056] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An impurity of Aprelet 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide, characterized in that: Its structural formula is shown in Formula I:

2. The method for preparing the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet according to claim 1, characterized in that: The following steps are involved: 1) using 7β-(N-tert-butyl-amino-formyl)androsta-3-bromo-3,5-diene as a raw material, reacting with a phosphine reagent under alkaline conditions in an organic solvent, and obtaining a crude reaction solution of 7β-(N-tert-butyl-amino-formyl)androsta-3-bromo-3,5-diene as a raw material ... reacting with a phosphine reagent under alkaline conditions in an organic solvent, and under halide catalysis; 2) The crude reaction solution of Aprelete impurity is concentrated by post-acid hydrolysis to obtain an oily substance, and the oily substance is purified by column chromatography to obtain Aprelete impurity; The reaction formula is as follows:

3. The method for preparing the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet according to claim 2, characterized in that: In the step 1), the mass ratio of the aplite Grignard compound, the phosphine reagent, the alkali reagent and the catalyst is 1:0.4-0.6:1.3-1.5:0.2-0.

6.

4. The method for preparing the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet according to claim 2, characterized in that: In the step 1), the mass volume ratio of the aplite Grignard compound to the organic solvent is 1:5-10.

5. The method for preparing the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet according to claim 2, characterized in that: The phosphine reagent in step 1) is diphenylphosphine oxide or triphenylphosphine.

6. The method for preparing the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet according to claim 2, characterized in that: The alkaline conditions in step 1) include at least one of dimethylethylenediamine, 1,8-diazabicycloundec-7-ene, triethylamine, cesium carbonate, sodium carbonate, potassium carbonate inorganic base or organic base, or a mixture of two thereof.

7. The method for preparing the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet according to claim 2, characterized in that: The catalyst in step 1) is at least one of cuprous iodide and sodium iodide, or a mixture of the two.

8. The method for preparing the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet according to claim 2, characterized in that: The organic solvent in step 1) includes at least one of toluene, tetrahydrofuran, dimethylformamide and DMSO.

9. The method for preparing the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Aprelet according to claim 2, characterized in that: The reaction temperature of step 1) is 100-110° C., and the reaction time is 20-48 hours.

10. Use of the impurity 17β-(N-tert-butyl-amino-formyl)androsta-3,5-diene-3-diphenylphosphine oxide of Apreletide according to claim 1 as a reference substance for quality research of Apreletide API and its preparations.

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