Method for removing d-component impurities to improve the quality of tilmicosin phosphate and application thereof
By adding an alkaline washing solution to the hydrolysis and reductive amination reaction solution of tilmicosin phosphate and adjusting the pH value to 10.0–11.0, the problem of high impurity content of D-component in tilmicosin phosphate was solved, thereby improving purity and production efficiency.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- ANHUI APELOA BIOTECHNOLOGY CO LTD
- Filing Date
- 2022-10-27
- Publication Date
- 2026-07-14
AI Technical Summary
Existing technologies struggle to effectively control the content of impurities in the D-component of tilmicosin phosphate, resulting in low product purity and low production efficiency.
The purity of tilmicosin phosphate was improved by adding an alkaline washing solution to the reaction solution after hydrolysis and reductive amination to adjust the pH to 10.0–11.0 and performing alkaline washing to remove impurities of the D-component. Combined with back-extraction and post-processing steps, the purity of tilmicosin phosphate was improved.
It effectively reduced the impurity content of the D-component from 4.5% to 1.5%, improving the purity and production efficiency of tilmicosin phosphate, simplifying the process and reducing costs.
Smart Images
Figure BDA0003912916180000011
Abstract
Description
Technical Field
[0001] This invention relates to a method for removing D-component impurities to improve the quality of tilmicosin phosphate, belonging to the field of biological veterinary drug preparation technology. Background Technology
[0002] Tilmicosin phosphate is a downstream product of tylosin, and its pharmacodynamic antibacterial activity is similar to that of tylosin, primarily against Gram-positive bacteria, but also effective against a few Gram-negative bacteria and mycoplasma. Its activity against Actinobacillus pleuropneumoniae, Pasteurella, and mycoplasma in livestock and poultry is stronger than that of tylosin. Its synthetic route uses tylosin as a starting material, and through reactions such as hydrolysis, reductive amination, and salt formation, tilmicosin phosphate is obtained.
[0003]
[0004] In industrial production, there are two main routes for the production of tilmicosin phosphate: one is the hydrolysis synthesis using tylosin salt, and the other is the direct hydrolysis synthesis using tylosin base. The hydrolysis synthesis using tylosin salt is more complex, with high energy consumption and low production efficiency. The direct hydrolysis synthesis using tylosin base, on the other hand, is difficult to control in terms of impurity levels, often resulting in excessively high levels of single impurities, especially those derived from the D-component. Because these compounds have similar properties to tilmicosin, existing processes are ineffective at removing impurities, easily exceeding pharmacopoeia standards. This often requires extensive purification operations, making the process complex and resulting in significant product loss. Summary of the Invention
[0005] After extensive research on impurities in tilmicosin phosphate, the inventors discovered that the main reason for the excessively high content of D-component impurities in tilmicosin phosphate is that the D-component content of tylosin is often higher than the pharmacopoeia standard. The excessively high D-component content of tylosin is mainly limited by the current fermentation and extraction technology of tylosin. Therefore, if the content of D-component impurities can be controlled at the source, it may be beneficial to improve the quality of tilmicosin phosphate and increase the production efficiency of tilmicosin phosphate.
[0006] To address the shortcomings of existing technologies, this invention provides a method for removing D-component impurities to improve the quality of tilmicosin phosphate. This method effectively removes impurities, improves the purity of tilmicosin phosphate, and is simple to operate and low in cost. Using this method, the D-component impurity level can be reduced from 4.5% to 1.5%.
[0007] The technical solution of the present invention is as follows:
[0008] A method for removing D-component impurities to improve the quality of tilmicosin phosphate includes the following steps:
[0009] (1) Add alkaline washing solution to the reaction solution after the hydrolysis of tylosin for alkaline washing. The amount of alkali added is such that the pH of the washing solution reaches 10.0 to 11.0. After washing, the alkali solution is drained to obtain the crude product of decarbonase sugar tylosin.
[0010] (2) After the crude decarbonase sugar product is reduced and amination, an alkaline washing solution is added to the reaction solution for alkaline washing. The amount of alkali added is such that the pH of the washing solution reaches 10.0-11.0. After washing, the alkaline solution is discharged, and then back-extraction is performed to obtain tilmicosin phosphate aqueous solution. After post-treatment, the tilmicosin phosphate is obtained.
[0011] The reaction solution obtained after the hydrolysis of tylosin is obtained by the following method:
[0012] The fermentation broth of tylosin was subjected to simple treatment, including flocculation filtration, ceramic membrane filtration, membrane concentration, and butyl acetate extraction. The resulting tylosin butyl acetate solution was then acid-extracted and directly hydrolyzed.
[0013] The principle of this invention is as follows:
[0014] This invention utilizes alkaline washing and hydrolysis to obtain tylosin, a decarbonase enzyme with high solubility in the organic phase. It also describes the reaction solution obtained after reductive amination. When the alkaline washing solution is mixed with the solvent phase, tilmicosin exhibits high solubility in the organic phase. Furthermore, due to the presence of dihydroxyl functional groups, the D-component impurities have a larger partition coefficient in the alkaline water. Therefore, alkaline washing of the hydrolysis reaction solution effectively removes the D-component impurities. In addition, the inventors discovered that both excessively high and low pH levels in the alkaline washing solution are ineffective in removing the D-component impurities. At excessively low pH, the partition coefficients of the D-component impurities are essentially the same in both the aqueous and organic phases. At excessively high pH, severe emulsification occurs in the washing solution, hindering the effective removal of the D-component.
[0015] Preferably, in step (1), after adding butyl acetate after the hydrolysis reaction, the alkaline washing solution is added for alkaline washing, and then extraction and separation are performed.
[0016] The alkaline washing solution is a 20-30% sodium hydroxide aqueous solution or potassium hydroxide aqueous solution.
[0017] As a further preferred option, the alkaline washing temperature is 20–30°C, and the alkaline washing time is 10–20 minutes.
[0018] Preferably, in step (2), the alkaline washing solution in the reaction solution after reduction amination is a sodium hydroxide aqueous solution or a potassium hydroxide aqueous solution with a concentration of 20-30%.
[0019] As a further preferred option, the alkaline washing temperature is 20–30°C, and the alkaline washing time is 10–20 minutes.
[0020] Preferably, in step (2), the mass ratio of the amount of alkaline washing solution added to the organic phase is 2-3:100.
[0021] Preferably, in step (2), after removing the alkali solution, water is added first, and then phosphoric acid is added for back-extraction. The pH of the back-extracted material is adjusted to 6.0-7.0.
[0022] Preferably, the volume ratio of water to organic phase is 1:1.8 to 1.2.
[0023] Preferably, the concentration of phosphoric acid is 75-85%.
[0024] Preferably, in step (2), the post-processing method includes decolorization, filtration, and spray drying.
[0025] The present invention also provides a method for preparing tilmicosin phosphate, using tylosin as the starting material, obtaining a hydrolyzed reaction solution through hydrolysis, and then obtaining tilmicosin phosphate by the above method.
[0026] The specific reaction processes of hydrolysis and reductive amination involved in this invention are existing technologies. Unless otherwise specified, those skilled in the art can make selections based on existing technologies.
[0027] Compared with the prior art, the present invention has the following beneficial effects:
[0028] (1) The present invention performs alkaline washing on the reaction solution after hydrolysis and reductive amination. After the alkaline washing solution is mixed with the solvent, the D-component impurities and the main products of the two-step reaction, namely decarbonase sugar tylosin and tilmicosin, have different partition coefficients in the aqueous phase and the organic phase. This allows some of the D-component impurities to be dissolved in the alkaline washing solution, while the main products are retained. The D-component impurities are also reduced, thereby achieving the goal of improving the quality of tilmicosin phosphate.
[0029] (2) The method of the present invention will not conflict with the existing tilmicosin phosphate production process, and is easy to implement. The raw materials required are also the raw materials that must be used to produce tilmicosin phosphate. Detailed Implementation
[0030] The present invention will be further described below through specific embodiments, but is not limited thereto.
[0031] Example 1
[0032] (1) Add 250 mL of water to 2500 g of tylosin butyl acetate solution (tylosin content is 2%), add 20% sulfuric acid to adjust the pH to 1.5-1.6, stir thoroughly, collect the aqueous phase, and carry out hydrolysis reaction at 35-45℃ for 2 h. Monitor the reaction by HPLC. After the raw material conversion is complete, add 150 mL of butyl acetate to the reaction solution and adjust the pH to 10-11 with 30% sodium hydroxide aqueous solution. Separate the solution and retain the organic phase. Add another 100 mL of butyl acetate to the aqueous phase for extraction, separate the phases, combine the two organic phases, concentrate until the solution is clear, and obtain the butyl acetate solution of decarbonylase tylosin.
[0033] After this step, the HPLC chromatographic purity of the main product and D-component impurities in the solution is as follows:
[0034] main product d-component (impurity) reaction solution 86％ 4.6％ concentrated solution 88％ 3.5％
[0035] (2) Add 6.32 g of 3,5-dimethylpiperidine and start heating with stirring. Heat to 65°C and begin adding 2.61 g of a 98% formic acid butyl acetate solution dropwise. The addition is completed in approximately 10 minutes. The temperature remains at 68-72°C. Continue the reaction at this temperature for 3 hours until the HPLC feedstock conversion is complete. Stop heating, cool to 30°C, add 100 mL of water, and adjust the pH of the solution to 10-11 with a 30% sodium hydroxide aqueous solution. Separate the solutions; wash the organic phase with purified water, separate the solutions, and retain the organic phase to obtain a butyl acetate solution of tilmicosin.
[0036] After this step, the HPLC chromatographic purity of the main product and D-component impurities in the solution is as follows:
[0037] main product d-component (impurity) reaction solution 90％ 3.5％ after washing 93％ 1.6％
[0038] (3) Add an equal volume of water to the above butyl acetate solution of tilmicosin, then add 5.5 g of 80% phosphoric acid aqueous solution, back-extract, collect the aqueous phase, add activated carbon for decolorization, filter, concentrate, spray dry, and obtain 45.6 g of tilmicosin phosphate solid, with a yield of 94%, a purity of 94.6%, and a maximum single impurity of 1.5%.
[0039] Example 2
[0040] (1) Add 250 mL of water to 2500 g of tylosin butyl acetate solution (tylosin content is 2%), add 20% sulfuric acid to adjust the pH to 1.5-1.6, stir thoroughly, collect the aqueous phase, and carry out hydrolysis reaction at 35-45℃ for 2 h. Monitor the reaction by HPLC. After the raw material conversion is complete, add 150 mL of butyl acetate to the reaction solution and adjust the pH to 9-10 with 30% sodium hydroxide aqueous solution. Separate the solution and retain the organic phase. Add another 100 mL of butyl acetate to the aqueous phase for extraction, separate the phases, combine the two organic phases, concentrate until the solution is clear, and obtain the butyl acetate solution of decarbonylase tylosin.
[0041] After this step, the HPLC chromatographic purity of the main product and D-component impurities in the solution is as follows:
[0042] main product d-component (impurity) reaction solution 86.6％ 4.6％ concentrated solution 87.1％ 4.2％
[0043] (2) Add 6.32 g of 3,5-dimethylpiperidine and start heating with stirring. Heat to 65°C and begin dropwise addition of 2.61 g of a 98% formic acid butyl acetate solution. The addition is completed in approximately 10 minutes. The temperature remains at 68-72°C. Continue the reaction at this temperature for 3 hours until the HPLC feedstock conversion is complete. Stop heating, cool to 30°C, add 100 mL of water, and adjust the pH of the solution to 9-10 with a 30% sodium hydroxide aqueous solution. Separate the solutions; wash the organic phase with purified water, separate the solutions, and retain the organic phase to obtain a butyl acetate solution of tilmicosin.
[0044] After this step, the HPLC chromatographic purity of the main product and D-component impurities in the solution is as follows:
[0045] main product d-component (impurity) reaction solution 88％ 4.0％ after washing 90％ 3.4％
[0046] (3) Add an equal volume of water to the above butyl acetate solution of tilmicosin, then add 5.5 g of 80% phosphoric acid aqueous solution, back-extract, collect the aqueous phase, add activated carbon for decolorization, filter, concentrate, spray dry, and obtain 46.1 g of tilmicosin phosphate solid, with a yield of 95%, a purity of 92.5%, and a maximum single impurity of 3.1%.
[0047] Example 3
[0048] (1) Add 250 mL of water to 2500 g of tylosin butyl acetate solution (tylosin content is 2%), add 20% sulfuric acid to adjust the pH to 1.5-1.6, stir thoroughly, collect the aqueous phase, and carry out hydrolysis reaction at 35-45℃ for 2 h. Monitor the reaction by HPLC. After the raw material conversion is complete, add 150 mL of butyl acetate to the reaction solution and adjust the pH to 9-10 with 30% sodium hydroxide aqueous solution. Separate the solution and retain the organic phase. Add another 100 mL of butyl acetate to the aqueous phase for extraction, separate the phases, combine the two organic phases, concentrate until the solution is clear, and obtain the butyl acetate solution of decarbonylase tylosin.
[0049] After this step, the HPLC chromatographic purity of the main product and D-component impurities in the solution is as follows:
[0050] main product d-component (impurity) reaction solution 86.9％ 4.7％ concentrated solution 87.3％ 4.5％
[0051] (2) Add 6.32 g of 3,5-dimethylpiperidine and start heating with stirring. Heat to 65°C and begin adding 2.61 g of a 98% formic acid butyl acetate solution dropwise. The addition is completed in approximately 10 minutes. The temperature remains at 68-72°C. Continue the reaction at this temperature for 3 hours until the HPLC feedstock conversion is complete. Stop heating, cool to 30°C, add 100 mL of water, and adjust the pH of the solution to 10-11 with a 30% sodium hydroxide aqueous solution. Separate the solutions; wash the organic phase with purified water, separate the solutions, and retain the organic phase to obtain a butyl acetate solution of tilmicosin.
[0052] After this step, the HPLC chromatographic purity of the main product and D-component impurities in the solution is as follows:
[0053] main product d-component (impurity) reaction solution 88％ 4.3％ after washing 90％ 3.2％
[0054] (3) Add an equal volume of water to the above butyl acetate solution of tilmicosin, then add 5.5 g of 80% phosphoric acid aqueous solution, back-extract, collect the aqueous phase, add activated carbon for decolorization, filter, concentrate, spray dry, and obtain 46.6 g of tilmicosin phosphate solid, with a yield of 96%, a purity of 93.1%, and a maximum single impurity of 3.5%.
[0055] Example 4
[0056] (1) Add 250 mL of water to 2500 g of tylosin butyl acetate solution (tylosin content is 2%), add 20% sulfuric acid to adjust the pH to 1.5-1.6, stir thoroughly, collect the aqueous phase, and carry out hydrolysis reaction at 35-45℃ for 2 h. Monitor the reaction by HPLC. After the raw material conversion is complete, add 150 mL of butyl acetate to the reaction solution and adjust the pH to 10-11 with 30% sodium hydroxide aqueous solution. Separate the solution and retain the organic phase. Add another 100 mL of butyl acetate to the aqueous phase for extraction, separate the phases, combine the two organic phases, concentrate until the solution is clear, and obtain the butyl acetate solution of decarbonylase tylosin.
[0057] After this step, the HPLC chromatographic purity of the main product and D-component impurities in the solution is as follows:
[0058] main product d-component (impurity) reaction solution 87.1％ 4.7％ concentrated solution 88.5％ 3.3％
[0059] (2) Add 6.32 g of 3,5-dimethylpiperidine and start heating with stirring. Heat to 65°C and begin dropwise addition of 2.61 g of a 98% formic acid butyl acetate solution. The addition is completed in approximately 10 minutes. The temperature remains at 68-72°C. Continue the reaction at this temperature for 3 hours until the HPLC feedstock conversion is complete. Stop heating, cool to 30°C, add 100 mL of water, and adjust the pH of the solution to 9-10 with a 30% sodium hydroxide aqueous solution. Separate the solutions; wash the organic phase with purified water, separate the solutions, and retain the organic phase to obtain a butyl acetate solution of tilmicosin.
[0060] After this step, the HPLC chromatographic purity of the main product and D-component impurities in the solution is as follows:
[0061] main product d-component (impurity) reaction solution 88.4％ 3.5％ after washing 90.1％ 2.8％
[0062] (3) Add an equal volume of water to the above tilmicosin butyl acetate solution, then add 5.5 g of 80% phosphoric acid aqueous solution, back-extract, collect the aqueous phase, add activated carbon for decolorization, filter, concentrate, spray dry, and obtain 46.1 g of tilmicosin phosphate solid, with a yield of 95%, a purity of 93.4%, and a maximum single impurity of 3.0%.
[0063] As can be seen from the four examples above, adding alkali to the hydrolysis reaction solution and the reductive amination reaction solution to adjust the pH to 10-11 for washing can significantly reduce the content of D-component impurities. Compared with the D-component content in the starting material, the alkali washing process can effectively reduce the D-component impurities by 2 percentage points, without loss of the main product or impact on the yield. This method requires washing the hydrolysis reaction solution and the reductive amination reaction solution separately. If only one reaction solution is washed, impurities cannot be effectively removed, resulting in substandard product purity. Furthermore, the pH value of the two washes must be between 10-11; too low a pH will not effectively remove impurities, while too high a pH will cause emulsification, making separation difficult.
Claims
1. A method for removing D-component impurities to improve the quality of tilmicosin phosphate, characterized in that, Includes the following steps: (1) Add alkaline washing solution to the reaction solution after the hydrolysis of tylosin for alkaline washing. The amount of alkali added is such that the pH of the washing solution reaches 10.0~11.
0. After washing, the alkali solution is drained to obtain the crude product of decarbonase sugar tylosin. (2) After the crude decarbonase sugar product is reduced and amination, an alkaline washing solution is added to the reaction solution for alkaline washing. The amount of alkali added is such that the pH of the washing solution reaches 10.0~11.
0. After washing, the alkaline solution is drained, and then back-extraction is performed to obtain tilmicosin phosphate aqueous solution. After post-treatment, the tilmicosin phosphate is obtained. The reaction solution after the hydrolysis of tylosin was obtained by the following method: The fermentation broth of tylosin was subjected to simple treatment, including flocculation filtration, ceramic membrane filtration, membrane concentration, and butyl acetate extraction. The resulting tylosin butyl acetate solution was then acid-extracted and directly hydrolyzed. In step (1), after the hydrolysis reaction, butyl acetate is added, followed by the addition of the alkaline washing solution for alkaline washing, and then extraction and separation are performed. The alkaline washing solution is a 20-30% sodium hydroxide aqueous solution or potassium hydroxide aqueous solution; The alkaline washing temperature is 20~30℃, and the alkaline washing time is 10~20 minutes; In step (2), the alkaline washing solution in the reaction solution after reduction amination is a sodium hydroxide aqueous solution or a potassium hydroxide aqueous solution with a concentration of 20-30%. The alkaline washing temperature is 20~30℃, and the alkaline washing time is 10~20 minutes; In step (2), the ratio of the amount of alkaline washing solution added to the mass of the organic phase is 2-3:
100.
2. The method for removing D-component impurities and improving the quality of tilmicosin phosphate according to claim 1, characterized in that, In step (2), after removing the alkali solution, water is added first, and then phosphoric acid is added for back-extraction. The pH of the back-extracted material is adjusted to 6.0-7.
0.
3. The method for removing D-component impurities and improving the quality of tilmicosin phosphate according to claim 2, characterized in that, The volume ratio of water to organic phase added is 1:1.8~1.2; The concentration of the phosphoric acid is 75-85%.
4. The method for removing D-component impurities and improving the quality of tilmicosin phosphate according to claim 1, characterized in that, In step (2), the post-processing method includes decolorization, filtration and spray drying.