Method for refining sinomenine hydrochloride for injection

Abstract

The invention aims to provide a refining process of a sinomenine hydrochloride bulk drug for injection, which comprises the following steps: taking 130-200 parts by volume of ethanol into a container, heating to reflux, adding the ethanol into a container containing 10 parts by weight of sinomenine hydrochloride, stirring, filtering while hot, cooling filtrate at 0-8 DEG C for crystallization, taking out after overnight, carrying out suction filtration, leaching filter residue with iced ethanol, and drying to obtain the sinomenine hydrochloride bulk drug for injection. And drying at 40 DEG C to obtain refined sinomenine hydrochloride. The refined sinomenine hydrochloride provided by the invention meets the impurity limit requirements of impurities (Q3A) in ICH new bulk drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine and relates to a new method for refining sinomenine hydrochloride for injection. Background Art

[0002] Sinomenine hydrochloride is a monomer compound extracted from Sinomenine sinomenine. The preparations made with it as the raw material include Zhengqing Fengtongning injection, Zhengqing Fengtongning sustained-release tablets, and Zhengqing Fengtongning tablets. The existing quality standards do not include the detection of related substances (impurities). In 2021, an analytical method for detecting impurities in Sinomenine hydrochloride was developed and established in accordance with the quality standards for chemical drugs. It was found that the main impurity of Sinomenine hydrochloride is bisinomenine. Through the detection of impurities in 10 batches of Sinomenine hydrochloride for injection produced under the existing process, their contents were all above the confirmation threshold (0.15%). According to ICH "Impurities in New Drug Substances" Q3A (12), impurities above the defined limit (0.15%) must be reduced to the confirmation threshold, so the refining process of Sinomenine hydrochloride raw material was studied. Summary of the invention

[0003] The purpose of the present invention is to optimize the refining process of raw materials for injection so that it meets the impurity limit requirements (Q3A) of impurities in new raw materials of ICH. The refining method of sinomenine hydrochloride for injection of the present invention is achieved by the following technical scheme.

[0004] Take 130-200 parts by volume of ethanol into a container and heat it to reflux, add it into a container containing 10 parts by weight of sinomenine hydrochloride, stir and filter while hot, cool the filtrate at 0-8°C for crystallization, take it out after overnight, filter it with suction, rinse the filter residue with ice ethanol, and dry it at 40°C to obtain refined sinomenine hydrochloride. The relationship between volume parts and weight parts in the present invention is the relationship between ml and g. BRIEF DESCRIPTION OF THE DRAWINGS

[0005] Figure 1 The content of double 1 in three batches of sinomenine hydrochloride before and after refining was %.

[0006] Figure 2 Double 2 trend chart of three batches of samples in the strong experiment.

[0007] Figure 3 Double 1 trend chart of three batches of samples in the strong experiment.

[0008] Figure 4 Trend chart of three batches of samples in the strong test with RT=0.94.

[0009] Figure 5 Trend chart of total impurity content in strong test.

[0010] The present invention optimizes the refining process of sinomenine hydrochloride injection raw material drug to make it meet the impurity limit requirements of ICH new raw materials (Q3A), and then prepares the experimental solution with the refined sample that meets the requirements according to the new prescription, and conducts high temperature strength test to examine its stability at high temperature. Experimental Example 1: Study on Refining Method 2.2.1 Refining method:

[0011] 2.2.1.1 Method 1 Take 20 ml of ethanol / methanol and heat it to reflux in a 100 ml round-bottom flask. Add it into a 100 ml beaker containing 1 g of sinomenine hydrochloride. After stirring, filter it while hot. Seal the filtrate and put it in a refrigerator at 0-8°C to cool and crystallize. Take it out after overnight, filter it with suction, rinse the residue with a small amount of ice ethanol / methanol, and dry it at 40°C for 1 hour. Weigh and calculate the yield. 200 mg of each batch is sent for related substance and content detection before and after purification.

[0012] 2.2.1.2 Method 2 Take 130ml of ethanol and 10g of sinomenine hydrochloride into a 250ml round-bottom flask and heat to reflux until the solution is clear. Filter while hot, and concentrate the filtrate at 50℃ under reduced pressure to 100%, 80%, 50%, and 30%. Seal it and place it in a refrigerator at 0-8℃ to cool and crystallize. Take it out after overnight, filter it with suction, rinse the residue with a small amount of ice ethanol, and dry it at 40℃ for 1h. Weigh to calculate the yield. 200mg of each batch before and after purification are tested for related substances and content.

[0013] 2.2.1.3 Method 3 Take 130 ml of ethanol and 10 g of sinomenine hydrochloride into a 250 ml round-bottom flask and heat under reflux until the solution is clear. Concentrate the solution at 50°C under reduced pressure to 100%, 80%, 50%, and 30%. Seal it and place it in a refrigerator at 0-8°C to cool and crystallize. Take it out after overnight, filter it, rinse the residue with a small amount of ice ethanol, and dry it at 40°C for 1 h. Calculate the yield by weighing. Send 200 mg of each batch before and after purification for related substance and content detection.

[0014] 2.2.1.4 Method 4 (original process) Take 10g of sinomenine hydrochloride, add it to a round-bottom flask containing 100ml of ethanol, stir and heat until boiling, and fully dissolve it. Continue heating, keep it slightly boiling, and perform vacuum concentration to recover ethanol until initial crystallization occurs. Seal it and place it in a refrigerator at 0-8℃ to cool and crystallize. Take it out after overnight, filter it, rinse the filter residue with a small amount of ice ethanol, and dry it at 40℃ for 1h to get it. Weigh and calculate the yield. 200mg of each batch is sent before and after refining for related substances and content detection.

[0015] 2.2.1.5 Method 5 Take 130ml of ethanol and put it into a 250ml round-bottom flask and heat it to reflux. Add it into a 250ml beaker containing 10g of sinomenine hydrochloride, stir and filter while hot. Seal the filtrate and put it into a refrigerator at 0-8℃ to cool and crystallize. Take it out after overnight, filter it with suction, rinse the residue with a small amount of ice ethanol, and dry it at 40℃ for 1h. Weigh and calculate the yield. 200mg of each batch is sent for related substance and content detection before and after refining. 2.2.2 Strong test

[0016] 2.2.2.1 Preparation of injection sample Take 100ml of injection water and heat it to 40℃, add 0.03g EDTA, add 2.5g sinomenine hydrochloride after dissolving, and then adjust the pH value to about 3 with 0.1mpl / L hydrochloric acid, take samples to determine the relevant substances, and then continue to reflux and heat to 100℃, reflux for 1h to obtain the injection sample solution 2.2.2.2 Strength test: Divide the sample solution into five parts and place them in a 70℃ stability test chamber. Take samples on the 0th, 1st, 3rd, 5th and 10th days (11 days at 70℃ is equivalent to 180 days at 40℃). [1] , determine the related substances.

[0017] 2.2.3 Related substances and content determination The related substances and content determination methods for injection in the internal control quality standard of sinomenine hydrochloride (QB / ZK-002-2023) are adopted. 2.2.3.1 Related substances

[0018] Test solution: Take an appropriate amount of the product, add 1% acetic acid to dissolve and dilute to make a solution containing about 5 mg per 1 ml.

[0019] Control solution: Accurately measure an appropriate amount of the test solution and quantitatively dilute it with 1% acetic acid to make a solution containing approximately 10 μg per 1 ml (0.2%).

[0020] Impurity A, Impurity B, Impurity C Reference Solution Take appropriate amount of Impurity A, Impurity B, Impurity C reference substances respectively, weigh accurately, add appropriate amount of 1% acetic acid to dissolve, and then dilute with 1% acetic acid to make reference substance solutions containing 0.02 mg per 1 ml.

[0021] System suitability solution: Take appropriate amounts of sinomenine hydrochloride, impurity A, impurity B, and impurity C reference substances and place them in the same volumetric flask. Add 1% acetic acid to dissolve them to make a solution containing 0.02 mg per 1 ml.

[0022] The chromatographic conditions were: octadecylsilane bonded silica gel as the filler (4.6 mm × 250 mm, 5 μm); sodium heptane sulfonate solution (1.01 g of sodium heptane sulfonate was dissolved in 1000 ml of water and the pH was adjusted to 2.5 ± 0.1 with phosphoric acid): methanol (90:10) as the mobile phase A, methanol: sodium heptane sulfonate solution (90:10) as the mobile phase B, and gradient elution was performed according to the table below; the flow rate was 1.5 ml per minute; the detection wavelength was 210 nm; the column temperature was 35 °C, and the injection volume was 20 μl.

[0023] 2.2.3.2 Content determination

[0024] The chromatographic conditions and system suitability test used octadecylsilane bonded silica gel without filler; acetonitrile-0.78% sodium dihydrogen phosphate solution (12:88) as the mobile phase; the detection wavelength was 265nm; and the theoretical plate number calculated based on the sinomenine peak should be no less than 2000.

[0025] Preparation of reference solution: Take about 20 mg of sinomenine reference substance dried to constant weight at 105°C, weigh accurately, place in a 100 ml volumetric flask, add 30 ml of ethanol, ultrasonically treat (power 300 W, frequency 25 kHz) for 15 minutes to dissolve, add mobile phase to dilute to scale, shake well, and make 1 ml containing 0.2 mg [equivalent to sinomenine hydrochloride (C 19 H 23 NO4·HCl) 0.222mg] solution is obtained.

[0026] Preparation of test solution: Take about 20 mg of the product, place it in a 100 ml volumetric flask, weigh it accurately, add 2 drops of ammonia test solution, add 30 ml of ethanol, and treat it ultrasonically (power 300 W, frequency 25 kHz) for 15 minutes to dissolve it. Cool it, add the mobile phase to dilute it to the scale, shake it well, filter it, and take the filtrate to obtain the solution.

[0027] Determination method: Accurately pipette 5μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0028] This product contains sinomenine hydrochloride (C 19 H 23 NO4·HCl) should not be less than 97.0%.

[0029] 3. Conventional instruments and reagents, as well as the test sample sinomenine hydrochloride API are all provided by the sample retention room of the Quality Management Department of Hunan Zhengqing Pharmaceutical Group Co., Ltd. 4.1. Results of related substances of eight batches of API in Method 1 Table 1 Impurities in eight batches of API

[0030]

[0031] The results showed that after the recrystallization treatment, the impurity A of the sinomenine hydrochloride raw material was reduced to below 0.05% except for batch number YK-220408, the impurity B was reduced to below 0.05%, and the impurity D was reduced to below 0.02% after methanol refining; after ethanol refining, the impurity A was reduced to below 0.03%, the impurity B was reduced to below 0.11%, and the impurity D was reduced to below 0.05%, which met the requirements of the Q3A (R2) definition decision tree of ICH "Impurities in New Drug Substances": impurities above the definition limit were reduced to below the definition limit (0.15%), and impurities above the identification limit were subjected to structural identification or reduced to below the identification limit (0.10%).

[0032] 4.2. Method Results of three batches of refinement experiments Table 2 Related substances of YK-220508

[0033] Table 3 Related substances of YK-220507

[0034] Table 4 Related substances of YK-220507

[0035]

[0036] Summary: According to the requirements of ICH for impurity limits in new APIs 1. The lowest content of double 1 in the three batches of refined samples prepared by Method 2 was 0.13% and the highest was 0.27%. Most of them exceeded the definition limit of 0.15% specified in ICH impurities in new drug substances (Q3A).

[0037] 2. The lowest content of double 1 in the three batches of refined samples prepared by Method 3 was 0.24% and the highest was 0.59%, both exceeding the definition limit of 0.15% specified in ICH impurities in new drug substances (Q3A).

[0038] 3. The content of double 1 in the three batches of refined samples of method 4 (original process) exceeded the definition limit of 0.15% specified in ICH impurities in new APIs (Q3A), with the lowest being 0.2% and the highest being 0.31%. 4. The impurity content of the three batches of refined samples of Method 5 was lower than the definition limit of 0.15% specified in ICH for impurities in new drug substances (Q3A), with the lowest being 0.07% and the highest being 0.09%.

[0039] 5. Regardless of the purification method used for the three batches of samples, except for double 1, the impurities of other impurities were all lower than 0.15%, which is in line with the impurity limits of ICH new drug substances (Q3A).

[0040] 6. The yields of the three batches of samples in method 5 are generally lower than those of other refining methods because the cooling rate of ethanol during the experiment is faster than that of others, resulting in a large amount of crystallization during filtration, resulting in a lower yield.

[0041] 4.3. Assay results Table 5 Content determination results

[0042]

[0043] Summary: The content of all samples is not less than 98% Table 6 Strong test results

[0044]

[0045] *“-” indicates the content is less than 0.02% Summary: From the above Figure 1-5 From Table 1-6, we can see 1. The total impurity content of the three batches of samples increased over time under the strong experimental conditions.

[0046] 2. The contents of bisinomenine 1 and the impurity with RT=0.94 in the three batches of samples were relatively stable during the 10-day high-temperature strength test, and were all below 0.15%.

[0047] 3. The content of sinomenine 2 in the three batches of samples increased with time under the strong experimental conditions. On the tenth day, the content of batch number YK-231007-10 exceeded 0.2%, reaching 0.36%.

[0048] 4. Therefore, it can be seen that high temperature has a significant impact on Zhengqing Fengtongning Injection, and it is recommended to store it at low temperature.

[0049] After screening, the impurity contents of sinomenine hydrochloride purified by method 5 all meet the impurity limit requirements of ICH new raw materials (Q3A). DETAILED DESCRIPTION

[0050] Example 1 Take 130ml-200ml of ethanol and heat it to reflux in a 250ml round-bottom flask. Add it into a 250ml beaker containing 10g of sinomenine hydrochloride, stir and filter while hot. Cool the filtrate at 0-8℃ to crystallize. Take it out after overnight, filter it with suction, rinse the residue with a small amount of ice ethanol, and dry it at 40℃. Example 2

[0051] Take 130L of ethanol and heat it to reflux in a 250L container. Add it to a 250L container containing 10kg of sinomenine hydrochloride. After stirring, filter it while hot. Cool the filtrate to 8°C for crystallization. Take it out after overnight, filter it with suction, rinse the residue with ice ethanol, and dry it at 40°C to obtain refined sinomenine hydrochloride. Example 3

[0052] Take 200L of ethanol and heat it to reflux in a 250L container. Add it to a 250L container containing 10kg of sinomenine hydrochloride. After stirring, filter it while hot. Cool the filtrate to 5°C for crystallization. Take it out after overnight, filter it with suction, rinse the residue with ice ethanol, and dry it at 40°C to obtain refined sinomenine hydrochloride. Example 4

[0053] Take 150ml of ethanol and heat it to reflux in a 250ml round-bottom flask. Add it into a 250ml beaker containing 10g of sinomenine hydrochloride, stir and filter while hot. Cool the filtrate at 0-8℃ to crystallize. Take it out after overnight, filter it with suction, rinse the residue with ice ethanol, and dry it at 40℃.

Claims

1. A method for refining sinomenine hydrochloride for injection, characterized in that: Take 130-200 parts by volume of ethanol into a container and heat it to reflux, add it into a container containing 10 parts by weight of sinomenine hydrochloride, stir and filter while hot, cool the filtrate at 0-8°C for crystallization, take it out after overnight, filter it with suction, rinse the residue with ice ethanol, and dry it at 40°C to obtain refined sinomenine hydrochloride.

2. The method for refining sinomenine hydrochloride for injection according to claim 1, characterized in that: Take 130 parts by volume of ethanol and heat it to reflux in a 250 parts by volume container, add it to a container containing 10 parts by weight of sinomenine hydrochloride, stir and filter while hot, cool the filtrate to 2°C for crystallization, take it out after overnight, filter it with suction, rinse the residue with ice ethanol, and dry it at 40°C to obtain refined sinomenine hydrochloride.

3. The method for refining sinomenine hydrochloride for injection according to claim 1, characterized in that: Take 130L of ethanol and heat it to reflux in a 250L container. Add it to a 250L container containing 10kg of sinomenine hydrochloride. After stirring, filter it while hot. Cool the filtrate to 8°C for crystallization. Take it out after overnight, filter it with suction, rinse the residue with ice ethanol, and dry it at 40°C to obtain refined sinomenine hydrochloride.

4. The method for refining sinomenine hydrochloride for injection according to claim 1, characterized in that: Take 200L of ethanol and heat it to reflux in a 250L container. Add it to a 250L container containing 10kg of sinomenine hydrochloride. After stirring, filter it while hot. Cool the filtrate to 5°C for crystallization. Take it out after overnight, filter it with suction, rinse the residue with ice ethanol, and dry it at 40°C to obtain refined sinomenine hydrochloride.

5. The method for refining sinomenine hydrochloride for injection according to claim 1, characterized in that: Take 130-120ml of ethanol and heat it to reflux in a 250ml round-bottom flask. Add it into a 250ml beaker containing 10g of sinomenine hydrochloride, stir and filter while hot. Cool the filtrate at 0-8℃ to crystallize. Take it out after overnight, filter it with suction, rinse the residue with ice ethanol, and dry it at 40℃.

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