Preparation method of umostat hydrogen sulfate

Abstract

The preparation method comprises the following steps: (1) mixing ethyl acetate, an intermediate II and sodium hydroxide, stirring until the mixture is clear, standing, carrying out liquid separation, and retaining an organic layer; (2) adding purified water into the organic layer obtained in the step (1), stirring, standing, separating liquid, and retaining the organic layer; (3) concentrating the organic layer obtained in the step (2) under reduced pressure until the organic layer is dry; (4) adding acetone into the concentrate obtained in the step (3), stirring and dissolving, filtering to remove insoluble substances, decompressing and concentrating the filtrate until the filtrate is dry, repeating the step, and stirring and dissolving after adding acetone for the last time; and (5) adding dilute sulphuric acid into the solution obtained in the step (4), heating to 55-75 DEG C, separating out solids, carrying out heat preservation and stirring for 1-3 hours, gradually cooling and stirring, and then discharging, centrifuging and drying to obtain the umostat hydrogen sulfate. Uromostat hydrogen sulfate can be chemically synthesized, the raw material source is stable, and extraction is convenient.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceuticals, and particularly to a preparation method of upamostat mesilate. Background Art

[0002] Upamostat (LH011) is an oral serine protease inhibitor. Serine proteases are a class of catalytically active enzymes that participate in a variety of physiological and pathological processes. In tumors, abnormally activated proteases can promote extracellular matrix degradation, cell migration, and angiogenesis, thus accelerating tumor progression. By specifically inhibiting serine proteases, it may block the key signaling pathways of tumor growth and metastasis, providing new molecular targets for cancer treatment. Traditional chemotherapy drugs can damage normal cells while killing tumor cells, resulting in serious side effects. Some targeted drugs may develop drug resistance after long-term use, limiting their clinical application. Developing oral drugs can improve patients' treatment compliance and quality of life.

[0003] As an oral serine protease inhibitor, upamostat can specifically inhibit the relevant proteases in the tumor microenvironment and block tumor progression. By optimizing the drug structure and administration route, upamostat aims to reduce toxic side effects and improve patients' tolerance. In addition to cancer treatment, the abnormal activation of serine proteases is also associated with various diseases (such as inflammation, fibrosis), and upamostat may have a wider range of indications. Currently, the preparation process of upamostat involves biological extraction and purification techniques, and the raw material sources are unstable, resulting in poor stability of the extraction process. Therefore, a chemical synthesis method is needed to prepare upamostat. Summary of the Invention

[0004] The purpose of the present invention is to provide a preparation method of upamostat mesilate, which can chemically synthesize upamostat mesilate with stable raw material sources and convenient extraction.

[0005] In one aspect of the present invention, a preparation method of upamostat mesilate is proposed. According to an embodiment of the present invention, the method includes the following steps:

[0006] (1) Mix ethyl acetate and Intermediate II, then add sodium hydroxide solution to adjust the pH to 7 - 8, stir until clear, then let stand and separate the layers, discard the lower aqueous layer, and retain the organic layer, wherein the chemical formula of Intermediate II is as follows:

[0007]

[0008] (2) Add purified water to the organic layer obtained in step (1), stir, then let stand and separate the layers, discard the lower aqueous layer, and retain the organic layer;

[0009] (3) Concentrate the organic layer obtained in step (2) under reduced pressure to dryness to obtain a concentrate.

[0010] (4) Add acetone to the concentrate obtained in step (3), stir to dissolve, filter to remove insoluble substances, concentrate the filtrate under reduced pressure to dryness, repeat this step 2 - 3 times, and stir to dissolve after adding acetone for the last time.

[0011] (5) Add dilute sulfuric acid to the solution in step (4), heat to 55 - 75 °C, solid will precipitate, keep stirring for 1 - 3 h while maintaining the temperature, cool to 30 - 50 °C, continue stirring for 1 - 5 h, then cool to 25 - 45 °C, stir for 10 - 20 h, then cool to 0 °C and continue stirring for 1 - 10 h, and then discharge the material and centrifuge to obtain the wet product of ulimorelin hydrogen sulfate.

[0012] (6) Dry the wet product of ulimorelin hydrogen sulfate to obtain the ulimorelin hydrogen sulfate.

[0013] In addition, according to the preparation method of ulimorelin hydrogen sulfate in the above - mentioned embodiments of the present invention, the following additional technical features may also be included:

[0014] In some embodiments of the present invention, in step (1), the mass ratio of intermediate II to ethyl acetate is 1:9.3 - 9.7.

[0015] In some embodiments of the present invention, in step (1), the mass percentage of the sodium hydroxide solution is 3.8% - 4%.

[0016] In some embodiments of the present invention, in step (2), the mass ratio of intermediate II to purified water is 1:5 - 5.5.

[0017] In some embodiments of the present invention, in step (4), the mass ratio of intermediate II to acetone is 1:1.5 - 1.7.

[0018] In some embodiments of the present invention, in step (5), the mass percentage of the dilute sulfuric acid is 45% - 65%

[0019] In some embodiments of the present invention, in step (6), the temperature of the drying is controlled at 30 - 50 °C, and drying is carried out under reduced pressure for 5 - 10 h.

[0020] Compared with the prior art, the beneficial effects of the present invention are:

[0021] (1) As a prodrug, the ulimorelin hydrogen sulfate prepared by the present invention, after intestinal absorption, the hydroxylamine group in its structure is reduced to an amino group, forming a pharmacologically active ingredient, a serine protease inhibitor, which inhibits the activity of serine protease, and then exerts an anti - tumor effect.

[0022] (2) The present invention successfully prepares ulinastatin hydrogen sulfate by chemical synthesis. Its raw material source is stable, and the components of the raw materials are more stable, providing a new idea for the synthesis of ulinastatin hydrogen sulfate. Detailed implementation mode

[0023] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Apparently, the described embodiments are only a part of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0024] Example 1

[0025] A preparation method of ulinastatin hydrogen sulfate, comprising the following steps:

[0026] (1) Prepare ulinastatin hydrogen sulfate intermediate I

[0027] a. Add 24.80 kg of dichloromethane to a 5-L enamel reaction kettle, start stirring, and sequentially add 2933.3 g of ethyl 4-[(2S)-2-amino-3-(3-cyanophenyl)propionyl]piperazinecarboxylate and 2416.0 g of 2,4,6-triisopropylbenzenesulfonyl chloride.

[0028] b. Slowly drop 1616.8 g of triethylamine into the reaction kettle, control the temperature at 30 °C, and stir and react for 20 hours.

[0029] c. Prepare two portions of dilute hydrochloric acid solution: Mix 786.7 g of hydrochloric acid and 7333.0 g of purified water.

[0030] d. Add one portion of the prepared dilute hydrochloric acid solution to the reaction kettle, stir, then turn off the stirring, let it stand, separate the liquid, and retain the lower organic layer.

[0031] e. Add the other portion of the prepared dilute hydrochloric acid solution to the organic layer, stir, then turn off the stirring, let it stand, separate the liquid, and retain the lower organic layer.

[0032] f. Add 8.00 kg of purified water to the organic layer, stir, then turn off the stirring, let it stand, separate the liquid, and retain the lower organic layer, and concentrate it under reduced pressure to dryness.

[0033] g. Add 6.40 kg of absolute ethanol to the concentrate, and continue to concentrate it under reduced pressure to dryness to obtain ulinastatin hydrogen sulfate intermediate I, and its chemical formula is as follows:

[0034]

[0035] The chemical reaction formula for the preparation of ulinastatin hydrogen sulfate intermediate I is as follows:

[0036]

[0037] (2) Preparation of Umaoxastat Hydrogen Sulfate Intermediate Ⅱ

[0038] a. Add the anhydrous ethanol solution of Intermediate Ⅰ into a 50 L glass-lined reactor, start stirring, and then add 11.05 kg of anhydrous ethanol.

[0039] b. Under nitrogen protection, add 726.6 g of 50% hydroxylamine solution, heat up to 60 °C, and stir for 10 hours.

[0040] c. After the reaction is completed, concentrate the reaction solution under reduced pressure to dryness. Then add 4.93 kg of ethyl acetate and concentrate to dryness under reduced pressure.

[0041] d. Add 37.70 kg of acetone to the concentrate and stir until it becomes clear.

[0042] e. Dissolve 1785.4 g of 1,5-naphthalenedisulfonic acid tetrahydrate in 4.26 kg of acetone.

[0043] f. Slowly add the acetone solution of 1,5-naphthalenedisulfonic acid tetrahydrate to the reactor. After the addition is complete, heat up to 20 °C and keep stirring for 2 hours.

[0044] g. Turn off the heating, cool down to 0 °C, and continue stirring for 10 hours.

[0045] h. Discharge the material, centrifuge, and obtain the wet product.

[0046] i. Add 20.92 kg of acetone to a 50 L glass-lined reactor, start stirring, add the wet product, and stir.

[0047] j. Discharge the material, centrifuge, and obtain the wet product of Intermediate Ⅱ.

[0048] k. Feed the wet product into a vacuum dryer, control the temperature at 30 °C, and dry under reduced pressure for 10 h. Obtain the white solid Intermediate Ⅱ, and its chemical formula is as follows:

[0049]

[0050] The chemical reaction formula for the preparation of Umaoxastat Hydrogen Sulfate Intermediate Ⅱ is as follows:

[0051]

[0052] (3) Preparation of Umaoxastat Hydrogen Sulfate

[0053] a. Prepare sodium hydroxide solution: Stir 1065.3 g of sodium hydroxide and 26.63 kg of purified water until completely dissolved, and cool to room temperature for standby.

[0054] b. Add 48.07 kg of ethyl acetate to a 100 L glass-lined reactor, start stirring, and add 5.06 kg of umostatin intermediate Ⅱ hydrogen sulfate. Drop the sodium hydroxide solution into the reactor and stir until it becomes clear. Check the pH = 7.

[0055] c. Turn off the stirring, let it stand, separate the liquid, discard the lower aqueous layer, and retain the organic layer.

[0056] d. Add 26.63 kg of purified water to the organic layer, stir, turn off the stirring, let it stand, separate the liquid, discard the lower aqueous layer, and retain the organic layer.

[0057] e. Concentrate the organic layer under reduced pressure to dryness.

[0058] f. After concentration, add 8.40 kg of acetone, stir to dissolve. Filter to remove insoluble substances. Transfer the filtrate to a 100 L glass-lined reactor and concentrate it to dryness under reduced pressure.

[0059] g. Add 20.90 kg of acetone to the concentrate, stir to dissolve, and concentrate it to dryness under reduced pressure.

[0060] h. Add 20.50 kg of acetone to the concentrate, stir to dissolve.

[0061] i. Dilute sulfuric acid: Slowly add 653.8 g of sulfuric acid along the wall of the container to 532.6 g of purified water while stirring.

[0062] j. Slowly drop the diluted sulfuric acid solution into the feed liquid. After dropping, raise the temperature to 65 °C. Solids will precipitate. Keep stirring for 2 h. Lower the temperature to 40 °C and continue stirring for 3 h. Lower the temperature to 35 °C and stir for 15 h, then lower the temperature to 0 °C and continue stirring for 5 h.

[0063] k. Discharge the material, centrifuge to obtain wet umostatin hydrogen sulfate.

[0064] l. Control the temperature at 40 °C and dry under reduced pressure for 8 hours to obtain the umostatin hydrogen sulfate.

[0065] The chemical reaction formula for the preparation of umostatin hydrogen sulfate is as follows:

[0066]

[0067] Example 2

[0068] The preparation method of umostatin hydrogen sulfate, the difference from Example 1 is only that: in step (3) j, slowly drop the diluted sulfuric acid solution into the feed liquid. After dropping, raise the temperature to 55 °C. Solids will precipitate. Keep stirring for 3 h. Lower the temperature to 30 °C and continue stirring for 5 h. Lower the temperature to 25 °C and stir for 20 h, then lower the temperature to 0 °C and continue stirring for 10 h.

[0069] Example 3

[0070] The preparation method of ulinastatin sulfate, which is only different from that of Example 1 in that: in step (3) j, the diluted sulfuric acid solution is slowly added dropwise to the feed liquid. After the addition is completed, the temperature is raised to 75 °C, and solids precipitate out. Keep stirring for 1 h. Cool down to 50 °C and continue stirring for 1 h. Cool down to 25 °C and stir for 10 h, and then cool down to 0 °C and continue stirring for 6 h.

[0071] The above content is only an example and illustration of the present invention. Those skilled in the art of this technology can make various modifications or supplements to the described specific embodiments or use similar methods for substitution. As long as it does not deviate from the structure of the present invention or exceed the scope defined by this claim book, it shall fall within the protection scope of the present invention.

Claims

1. A preparation method of ulinastatin bisulfate, characterized in that, It includes the following steps: (1) Mix ethyl acetate and Intermediate II, then add sodium hydroxide solution to adjust the pH to 7 - 8, stir until clear, then let it stand, separate the layers, discard the lower aqueous layer, and retain the organic layer. The chemical formula of Intermediate II is as follows: (2) Add purified water to the organic layer obtained in step (1), stir, then let it stand, separate the layers, discard the lower aqueous layer, and retain the organic layer; (3) Concentrate the organic layer obtained in step (2) under reduced pressure to dryness to obtain a concentrate; (4) Add acetone to the concentrate obtained in step (3), stir to dissolve, filter to remove insoluble substances, concentrate the filtrate under reduced pressure to dryness, repeat this step 2 - 3 times, and finally stir to dissolve after adding acetone; (5) Add dilute sulfuric acid to the solution in step (4), heat up to 55 - 75 °C, solid will precipitate, keep stirring for 1 - 3 h, cool down to 30 - 50 °C, continue stirring for 1 - 5 h, then cool down to 25 - 45 °C, stir for 10 - 20 h, then cool down to 0 °C and continue stirring for 1 - 10 h, then discharge the material and centrifuge to obtain the wet umostatin sulfate; (6) Dry the wet umostatin sulfate to obtain the umostatin sulfate.

2. The preparation method of ulimorelin hydrogen sulfate according to claim 1, characterized in that: In step (1), the mass ratio of Intermediate II to ethyl acetate is 1:9.3 - 9.

7.

3. The preparation method of ulimorelin hydrogen sulfate according to claim 1, characterized in that: In step (1), the mass percentage of the sodium hydroxide solution is 3.8% - 4%.

4. The preparation method of ulimorelin hydrogen sulfate according to claim 1, characterized in that: In step (2), the mass ratio of Intermediate II to purified water is 1:5 - 5.

5.

5. The preparation method of ulimorelin hydrogensulfate according to claim 1, characterized in that: In step (4), the mass ratio of Intermediate II to acetone is 1:1.5 - 1.

7.

6. The preparation method of ulinastatin hydrogen sulfate according to claim 1, characterized in that: In step (5), the mass percentage of the dilute sulfuric acid is 45% - 65%.

7. The preparation method of ulimorelin hydrogen sulfate according to claim 1, characterized in that: In step (6), the drying temperature is controlled at 30 - 50 °C, and vacuum drying is carried out for 5 - 10 h.

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