17α-Hydroxyprogesterone and its preparation method

Abstract

The present invention belongs to the technical field of progesterone preparation, and discloses a 17α-hydroxyprogesterone and a preparation method thereof. The present invention provides a preparation method of 17α-hydroxyprogesterone. The 17α-hydroxyprogesterone is obtained by the method combining chemical method and enzymatic method, specifically: progesterone is obtained through the carbonyl protection of dihydrotachysterol, catalytic oxidation, decarbonylation oxidation and deprotection, and then 17α-hydroxyprogesterone is obtained by the hydroxylation reaction of progesterone and 17α-hydroxylase. The preparation process of the present invention is simple, the raw material cost is low, the reaction temperature is low, and the reaction time is short. The present invention also provides the 17α-hydroxyprogesterone obtained by the above preparation method. The yield and purity of the 17α-hydroxyprogesterone obtained by the present invention are improved. The purity of the 17α-hydroxyprogesterone is above 97.5%, and the yield reaches above 90%. It lays a foundation for the development of the preparation of 17α-hydroxyprogesterone with high purity and high yield.

Description

Technical Field

[0001] The present invention relates to the technical field of progesterone preparation, and in particular to a 17α-hydroxyprogesterone and a preparation method thereof. Background Art

[0002] 17α-hydroxyprogesterone is an intermediate required for synthesizing hormonal drugs. 17α-hydroxyprogesterone is a key intermediate for synthesizing many steroid drugs such as medroxyprogesterone acetate, prednisolone, betamethasone, and dexamethasone.

[0003] Currently, 17α-hydroxyprogesterone is mainly prepared from 4-androstenedione. However, this method has the problems that the quality of the produced 17α-hydroxyprogesterone is unstable and cannot meet the requirements of higher-performance applications. Moreover, the above process needs to go through steps such as hydroxynitrification, dehydration, carboxy-ketone protection, hydrogenation, and addition hydrolysis. The preparation process is complicated, time-consuming, and requires a high reaction temperature. For example, Chinese Patent CN103524588 discloses a method for preparing progesterone. The preparation process needs to go through steps such as hydroxynitrification, dehydration, carboxy-ketone protection, hydrogenation, and addition hydrolysis. The required reaction temperature is about 100°C, and the reaction takes about 30 hours, which greatly reduces the production efficiency. Therefore, it is urgent in this field to develop a preparation method of 17α-hydroxyprogesterone with short time consumption, low energy consumption, and high yield. Summary of the Invention

[0004] In view of this, the present invention provides a 17α-hydroxyprogesterone and a preparation method thereof to solve the problems that the existing preparation of 17α-hydroxyprogesterone has a complicated preparation process, is time-consuming, and requires a high reaction temperature.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] The present invention provides a method for preparing 17α-hydroxyprogesterone, comprising the following steps:

[0007] (1) Mixing bisnorchol, ethylene glycol, and a catalyst to obtain bisnorchol with a protected carbonyl group;

[0008] (2) Mixing bisnorchol with a protected carbonyl group, an acidic ionic liquid, a heteropolyacid, and an oxidant, and reacting to obtain an enone intermediate;

[0009] (3) Mixing the enone intermediate, an acidic ionic liquid, and a solvent, and reacting to obtain progesterone;

[0010] (4) Mixing progesterone, 17α-hydroxylase, a coenzyme, and a regenerase, and hydroxylating to obtain 17α-hydroxyprogesterone.

[0011] Preferably, in the step (1), the catalyst is hydrochloric acid or sulfuric acid; the mass ratio of bicyclohexanol to ethylene glycol is 50-60:1-2; the mass ratio of the catalyst to ethylene glycol is 0.3-0.6:1.

[0012] Preferably, the acidic ionic liquid is a polyvinyl alcohol acidic ionic liquid; the mass of the acidic ionic liquid in step (2) and the mass of the acidic ionic liquid in step (3) are independently 2-3% of the mass of bicyclohexanol.

[0013] Preferably, in the step (2), the heteropolyacid is phosphomolybdic heteropolyacid; the oxidant is hydrogen peroxide or perchloric acid; the mass ratio of the acidic ionic liquid, the heteropolyacid and the oxidant is 1:10-15:5-8.

[0014] Preferably, in the step (2), the reaction temperature is 40-50 °C and the reaction time is 5-6 h.

[0015] Preferably, in the step (3), the solvent is methanol, isopropanol or n-butanol; the mass-volume ratio of the acidic ionic liquid to the solvent is 1 g:50-80 mL.

[0016] Preferably, in the step (3), the reaction temperature is 50-60 °C and the reaction time is 2-4 h.

[0017] Preferably, in the step (4), the coenzyme is riboflavin or thiamine; the regenerase is formate dehydrogenase, glucose dehydrogenase or hydrogenase; the dosage of 17α-hydroxylase is 10-20% of the mass of progesterone; the mass ratio of 17α-hydroxylase, coenzyme and regenerase is 1:0.2-0.5:0.3-0.6.

[0018] Preferably, in the step (4), the hydroxylation temperature is 20-30 °C, the hydroxylation time is 2-3 h, and the hydroxylation pH value is 6.8-7.2.

[0019] The present invention also provides 17α-hydroxyprogesterone prepared by the preparation method of 17α-hydroxyprogesterone.

[0020] It can be seen from the above technical solutions that compared with the prior art, the beneficial effects of the present invention are as follows:

[0021] The present invention prepares 17α-hydroxyprogesterone by combining chemistry and enzymatic method, with short reaction time required, low reaction temperature required, reducing energy consumption; at the same time, the raw materials of the present invention are simple and easy to obtain, reducing the reaction cost; and the purity and yield of 17α-hydroxyprogesterone obtained by the present invention are high. Detailed Embodiments

[0022] The present invention provides a preparation method of 17α-hydroxyprogesterone, comprising the following steps:

[0023] (1) Mix bicyclo[2.2.1]heptan-2-ol, ethylene glycol and a catalyst to obtain bicyclo[2.2.1]heptan-2-ol with a protected carbonyl group;

[0024] (2) Mix bicyclo[2.2.1]heptan-2-ol with a protected carbonyl group, an acidic ionic liquid, a heteropolyacid and an oxidant, and react to obtain an enone intermediate;

[0025] (3) Mix the enone intermediate, an acidic ionic liquid and a solvent, and react to obtain progesterone;

[0026] (4) Mix progesterone, 17α-hydroxylase, a coenzyme and a regenerase, and hydroxylate to obtain 17α-hydroxyprogesterone.

[0027] In the present invention, in the step (1), the catalyst is preferably hydrochloric acid or sulfuric acid, more preferably hydrochloric acid; the mass fraction of hydrochloric acid is preferably 45-50%, more preferably 46-48%; the mass fraction of sulfuric acid is preferably 40-60%, more preferably 45-55%; the mass ratio of bicyclo[2.2.1]heptan-2-ol to ethylene glycol is preferably 50-60:1-2, more preferably 52-55:1.2-1.6; the mass ratio of the catalyst to ethylene glycol is preferably 0.3-0.6:1, more preferably 0.4-0.5:1.

[0028] In the present invention, the acidic ionic liquid is preferably a polyethylene glycol acidic ionic liquid; the mass of the acidic ionic liquid in step (2) and the mass of the acidic ionic liquid in step (3) are independently preferably 2-3% of the mass of bicyclo[2.2.1]heptan-2-ol, more preferably 2.5-2.8% of the mass of bicyclo[2.2.1]heptan-2-ol;

[0029] The structural formula of the polyethylene glycol acidic ionic liquid is as follows:

[0030]

[0031] In the present invention, in the step (1), before obtaining bicyclo[2.2.1]heptan-2-ol with a protected carbonyl group, subject the mixed product to vacuum distillation; the pressure of the vacuum distillation is preferably 10-30 kPa, more preferably 15-25 kPa; the temperature of the vacuum distillation is preferably 60-100 °C, more preferably 75-85 °C.

[0032] In the present invention, the reaction formula of the step (1) is as follows:

[0033]

[0034] In the present invention, in the step (2), the heteropoly acid is preferably phosphomolybdic heteropoly acid; the oxidant is preferably hydrogen peroxide or perchloric acid; the concentration of hydrogen peroxide is preferably 30-40%, more preferably 32-38%; the concentration of perchloric acid is preferably 50-72%, more preferably 60-70%; the mass ratio of the acidic ionic liquid, the heteropoly acid and the oxidant is preferably 1:10-15:5-8, more preferably 1:11-14:6-7.

[0035] In the present invention, in the step (2), the reaction temperature is preferably 40-50°C, more preferably 45-48°C, and the reaction time is preferably 5-6 h, more preferably 5.5 h.

[0036] In the present invention, in the step (2), after the reaction is completed, the obtained product is sequentially concentrated, dried and filtered by suction to obtain the enone intermediate;

[0037] The temperature of concentration and drying is preferably 70-100°C, more preferably 80-90°C.

[0038] In the present invention, the reaction formula of the step (2) is as follows:

[0039]

[0040] In the present invention, in the step (3), the solvent is preferably methanol, isopropanol or n-butanol, more preferably methanol or n-butanol; the mass-to-volume ratio of the acidic ionic liquid and the solvent is preferably 1 g:50-80 mL, more preferably 1 g:60-70 mL.

[0041] In the present invention, in the step (3), the reaction temperature is preferably 50-60°C, more preferably 55-58°C; the reaction time is preferably 2-4 h, more preferably 3-3.5 h.

[0042] In the present invention, the reaction formula of the step (3) is as follows:

[0043]

[0044] In the present invention, in the step (3), after the reaction is completed, the reaction product is mixed with sulfuric acid to remove the protecting group, and then cooled to room temperature. After standing for 1-3 h, it is filtered by suction to obtain progesterone;

[0045] The mass-volume ratio of the product obtained from the reaction to sulfuric acid is preferably 1 g: 10-20 mL, more preferably 1 g: 12-15 mL; the mass fraction of sulfuric acid is preferably 40-60%, more preferably 45-55%; the mixing time is preferably 10-30 min, more preferably 15-25 min; the standing time is preferably 1.5-2.5 h, more preferably 2 h.

[0046] In the present invention, in the step (4), the coenzyme is preferably riboflavin or thiamine, more preferably riboflavin; the regenerase is preferably formate dehydrogenase, glucose dehydrogenase or hydrogenase, more preferably glucose dehydrogenase; the dosage of 17α-hydroxylase is preferably 10-20% of the mass of progesterone, more preferably 12-18% of the mass of progesterone; the mass ratio of 17α-hydroxylase, coenzyme and regenerase is preferably 1: 0.2-0.5: 0.3-0.6, more preferably 1: 0.3-0.4: 0.4-0.5.

[0047] In the present invention, in the step (4), the hydroxylation temperature is preferably 20-30 °C, more preferably 25-26 °C; the hydroxylation time is preferably 2-3 h, more preferably 2.5 h; the pH value of hydroxylation is preferably 6.8-7.2, more preferably 7.

[0048] In the present invention, in the step (4), after the hydroxylation is completed, the obtained product is washed successively with sulfuric acid and water, and then dried to obtain 17α-hydroxyprogesterone;

[0049] The number of washing times is preferably 2-4 times, more preferably 3 times; the drying temperature is preferably 70-90 °C, more preferably 75-85 °C.

[0050] The present invention also provides 17α-hydroxyprogesterone prepared by the preparation method of the 17α-hydroxyprogesterone.

[0051] The technical solutions provided by the present invention will be described in detail below with reference to the examples, but they should not be construed as limiting the protection scope of the present invention.

[0052] Example 1

[0053] (1) Add 100 g of dihydroxydrogesterone, 2 g of ethylene glycol and 0.6 g of sulfuric acid (mass fraction 45%) to a beaker and mix for 20 min, then perform vacuum distillation at 20 kPa and 80 °C to obtain dihydroxydrogesterone with the carbonyl group protected;

[0054] (2) Add the carbonyl-protected bisnorbornanol obtained in step (1), 2 g of polyvinyl alcohol acidic ionic liquid, 20 g of phosphomolybdic heteropolyacid, and 11 g of hydrogen peroxide (concentration: 35%) to a three-necked flask and mix them. Under stirring conditions, react at 40 °C for 5 h, then concentrate and dry at 80 °C, and then filter to obtain the enone intermediate;

[0055] (3) Add the enone intermediate obtained in step (2), 2.1 g of polyvinyl alcohol acidic ionic liquid, and 105 mL of methanol to a beaker and mix them. React at 50 °C for 2 h, then add 100 mL of sulfuric acid (mass fraction: 40%), stir for 20 min, cool to room temperature, let stand for 1 h, and then filter to obtain progesterone;

[0056] (4) Mix 6 g of progesterone obtained in step (3), 1.2 g of 17α-hydroxylase, 0.24 g of thiamine, and 0.36 g of formate dehydrogenase, and hydroxylate at 25 °C and pH 7 for 2 h to obtain 17α-hydroxyprogesterone.

[0057] Detect and calculate the yield and purity of the 17α-hydroxyprogesterone obtained in this example. The yield of the 17α-hydroxyprogesterone obtained in this example is 90%, and the purity is 98%.

[0058] Example 2

[0059] (1) Add 110 g of bisnorbornanol, 2.1 g of ethylene glycol, and 0.84 g of sulfuric acid (mass fraction: 45%) to a beaker and mix them for 20 min, then perform vacuum distillation at 22 kPa and 85 °C to obtain the carbonyl-protected bisnorbornanol;

[0060] (2) Add the carbonyl-protected bisnorbornanol obtained in step (1), 2.2 g of polyvinyl alcohol acidic ionic liquid, 23 g of phosphomolybdic heteropolyacid, and 13.2 g of hydrogen peroxide (concentration: 35%) to a three-necked flask and mix them. Under stirring conditions, react at 45 °C for 5 h, then concentrate and dry at 80 °C, and then filter to obtain the enone intermediate;

[0061] (3) Add the enone intermediate obtained in step (2), 2.3 g of polyvinyl alcohol acidic ionic liquid, and 115 mL of methanol to a beaker and mix them. React at 55 °C for 3 h, then add 105 mL of sulfuric acid (mass fraction: 45%), stir for 20 min, cool to room temperature, let stand for 1 h, and then filter to obtain progesterone;

[0062] (4) Mix 7.5 g of progesterone obtained in step (3), 1.5 g of 17α-hydroxylase, 0.6 g of thiamine, and 0.6 g of formate dehydrogenase, and hydroxylate at 25 °C and pH 7 for 2.5 h to obtain 17α-hydroxyprogesterone.

[0063] The yield and purity of 17α-hydroxyprogesterone obtained in this example were detected and calculated. The yield of 17α-hydroxyprogesterone obtained in this example was 90.2%, and the purity was 97.8%.

[0064] Example 3

[0065] (1) Add 90 g of bishomohydroxysterol, 1.5 g of ethylene glycol, and 0.9 g of sulfuric acid (mass fraction of 45%) to a beaker and mix for 20 min. Then, perform vacuum distillation at 20 kPa and 80 °C to obtain bishomohydroxysterol with a protected carbonyl group;

[0066] (2) Add the bishomohydroxysterol with a protected carbonyl group obtained in step (1), 1.8 g of polyvinyl alcohol acidic ionic liquid, 18 g of phosphomolybdic heteropolyacid, and 12.6 g of hydrogen peroxide (concentration of 35%) to a three-necked flask and mix. Under stirring conditions, react at 48 °C for 6 h, then concentrate and dry at 80 °C, and then perform suction filtration to obtain an enone intermediate;

[0067] (3) Add the enone intermediate obtained in step (2), 1.85 g of polyvinyl alcohol acidic ionic liquid, and 90 mL of methanol to a beaker and react at 60 °C for 3 h. Then, add 98 mL of sulfuric acid (mass fraction of 45%), stir for 20 min, and then cool to room temperature. Let it stand for 1 h and then perform suction filtration to obtain progesterone;

[0068] (4) Mix 5.5 g of progesterone obtained in step (3), 1.1 g of 17α-hydroxylase, 0.22 g of thiamine, and 0.44 g of formate dehydrogenase, and hydroxylate at 25 °C and pH 7 for 3 h to obtain 17α-hydroxyprogesterone.

[0069] The yield and purity of 17α-hydroxyprogesterone obtained in this example were detected and calculated. The yield of 17α-hydroxyprogesterone obtained in this example was 91.2%, and the purity was 97.5%.

[0070] It can be seen from Examples 1 to 3 that the present invention synthesizes 17α-hydroxyprogesterone by a chemo-enzymatic method, simplifies the process steps, reduces the reaction temperature and time, and reduces the reaction energy consumption. At the same time, the purity and yield of the finally obtained 17α-hydroxyprogesterone are improved. The purity of 17α-hydroxyprogesterone obtained in the present invention is above 97.5%, and the yield reaches above 90%.

[0071] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A method for preparing 17α-hydroxyprogesterone, characterized in that, It includes the following steps: (1) Mix bicyclo[2.2.1]hept-2-ene-7,7-dimethanol, ethylene glycol and a catalyst to obtain bicyclo[2.2.1]hept-2-ene-7,7-dimethanol with protected carbonyl group; (2) Mix bicyclo[2.2.1]hept-2-ene-7,7-dimethanol with protected carbonyl group, an acidic ionic liquid, a heteropolyacid and an oxidant, and react to obtain an enone intermediate; (3) Mix the enone intermediate, the acidic ionic liquid and a solvent, and react to obtain progesterone; (4) Mix progesterone, 17α-hydroxylase, coenzyme and regenerase, and hydroxylate to obtain 17α-hydroxyprogesterone; The acidic ionic liquid is a polyethylene glycol acidic ionic liquid, and the structural formula is: ； In the step (2), the heteropolyacid is phosphomolybdic heteropolyacid.

2. The preparation method of 17α-hydroxyprogesterone according to claim 1, characterized in that, In the step (1), the catalyst is hydrochloric acid or sulfuric acid; the mass ratio of bicyclo[2.2.1]hept-2-ene-7,7-dimethanol to ethylene glycol is 50-60:1-2; the mass ratio of the catalyst to ethylene glycol is 0.3-0.6:

1.

3. The preparation method of 17α-hydroxyprogesterone according to claim 2, wherein, The mass of the acidic ionic liquid described in the step (2) and the mass of the acidic ionic liquid described in the step (3) are independently 2-3% of the mass of bicyclo[2.2.1]hept-2-ene-7,7-dimethanol.

4. The preparation method of 17α-hydroxyprogesterone according to any one of claims 1 to 3, characterized in that, In the step (2), the oxidant is hydrogen peroxide or perchloric acid; the mass ratio of the acidic ionic liquid, the heteropolyacid and the oxidant is 1:10-15:5-8.

5. The preparation method of 17α-hydroxyprogesterone according to claim 4, characterized in that, In the step (2), the reaction temperature is 40-50 °C, and the reaction time is 5-6 h.

6. The preparation method of 17α-hydroxyprogesterone according to claim 1 or 5, characterized in that, In the step (3), the solvent is methanol, isopropanol or n-butanol; the mass-volume ratio of the acidic ionic liquid to the solvent is 1 g:50-80 mL.

7. The preparation method of 17α-hydroxyprogesterone according to claim 6, wherein, In the step (3), the reaction temperature is 50-60 °C, and the reaction time is 2-4 h.

8. The preparation method of 17α-hydroxyprogesterone according to claim 7, characterized in that, In the step (4), the coenzyme is riboflavin or thiamine; the regenerase is formate dehydrogenase, glucose dehydrogenase or hydrogenase; the dosage of 17α-hydroxylase is 10-20% of the mass of progesterone; the mass ratio of 17α-hydroxylase, coenzyme and regenerase is 1:0.2-0.5:0.3-0.

6.

9. The preparation method of 17α-hydroxyprogesterone according to claim 8, wherein, In the step (4), the hydroxylation temperature is 20-30 °C, the hydroxylation time is 2-3 h, and the hydroxylation pH value is 6.8-7.2.

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