Preparation method of lyophilized preparation of thymalfasin for injection

Abstract

The invention discloses a preparation method of a lyophilized preparation of thymalfasin for injection, and relates to a preparation method of a lyophilized preparation in the field of medicines. According to the method, thymalfasin, mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate are used as raw and auxiliary materials to prepare the thymalfasin freeze-dried preparation for injection, after split charging and half plugging, a new freeze-drying curve is adopted for freeze-drying treatment, and the freeze-drying curve improves energy supply during one-time sublimation, so that the one-time sublimation process is obviously accelerated; the freeze-drying process time is shortened; meanwhile, the product quality is ensured. According to the thymalfasin freeze-dried powder for injection produced by the method disclosed by the invention, the freeze-drying period is obviously shortened, the mass commercial amplification quality is consistent, and the thymalfasin freeze-dried powder has obvious commercial value.

Description

Technical Field

[0001] The invention relates to a method for preparing a freeze-dried preparation in the medical field, and in particular to a method for preparing a freeze-dried preparation of thymalfasin for injection. Background Art

[0002] Thymosin is a protein and polypeptide hormone produced by the thymus of animals, and thymosin is a high-end product among thymosin. It is the main biologically active component of thymosin and an important immunomodulatory substance in the human body. The original thymosin injection formulation was developed by SciClone Pharmaceuticals Italy SRL and is marketed under the trade name (Chinese name ), with a specification of 1.6mg. It is clinically used as a vaccine enhancer for the treatment of chronic hepatitis B and immune-compromised patients. The original product has been approved for marketing in China.

[0003] Chinese patent document CN 115998849A discloses a method for preparing thymosin lyophilized powder, comprising the following steps: firstly preparing a liquid medicine, taking 500 mg of thiomannitol according to the formula ratio, dissolving it in 50 g (10-30° C.) water for injection, stirring and dissolving, then adding 160 mg of thymosin to dissolve it, slowly adding a phosphate buffer solution (sodium dihydrogen phosphate and disodium hydrogen phosphate are added in appropriate amounts and mixed) aqueous solution, adjusting the pH value to 6.5-7.0, adding water for injection to 100 g, packaging, 1 ml / bottle, half-stoppering; preparing a liquid medicine to be lyophilized; the process also includes preparing the drug of the above formula into lyophilized powder, and performing the following treatments:

[0004] a: Place the qualified products on the freeze dryer partition, and keep the product temperature at -35-40℃;

[0005] b: After the freeze-dried product is packed, keep this temperature for 2-3 hours;

[0006] c: The condenser is cooled to below -50°C, and the vacuum is evacuated to a vacuum degree of ≤0.3mbar. The temperature rise program is set, and the shelf temperature is raised to -30°C in 2 hours, and the shelf temperature is maintained for 2-4 hours;

[0007] d: The shelf temperature is raised to -15°C over a period of 2 hours and the shelf temperature is maintained for 10 hours;

[0008] e: The shelf temperature is raised to 15°C in 1 hour;

[0009] f: Raise the shelf temperature to 40°C in 1 hour, maintain for 5-8 hours, press the stopper and take it out of the freeze dryer.

[0010] This patented process is mainly carried out at -15℃, which takes a long time, and the quality of the freeze-dried products cannot be guaranteed when the batch is enlarged, and it is difficult to reproduce. The existing process has problems such as long freeze-drying time, high energy consumption, high moisture content of the product, and slightly poor quality of the product during long-term storage. Summary of the invention

[0011] The object of the present invention is to provide a new method for preparing a lyophilized preparation of thymosin for injection in view of the deficiencies in the prior art. The method has good stability and low energy consumption, can significantly shorten the lyophilization cycle, save electricity, reduce costs, and is suitable for large-scale production.

[0012] The technical problem to be solved by the present invention is to provide a new method for preparing a lyophilized preparation of thymosin, with the following characteristics:

[0013] The active ingredients of each 1000mL of thymosin freeze-dried preparation for injection are:

[0014] Thymosin 1000-2200mg, mannitol 20-70g, sodium dihydrogen phosphate, sodium hydrogen phosphate as appropriate;

[0015] Then add water for injection to 1000ml;

[0016] pH is 6.8-7.3; packaging, half-stoppering to obtain the lyophilized drug solution;

[0017] The freeze-drying process was performed using the following freeze-drying curve:

[0018] ① Place the qualified products on the freeze dryer shelf, and keep the freeze dryer shelf temperature at 5℃~22℃;

[0019] ②After packing, keep the plate temperature at -40℃ or below, pre-freeze and cool down for 30 to 90 minutes until the product temperature is -35 to -45℃, and keep this temperature for 1 to 4 hours;

[0020] ③Raise the shelf temperature quickly to 0℃~5℃ and maintain the shelf temperature for 4~6 hours; when the product temperature reaches -10℃~-7℃, drop the shelf temperature to -5℃ and maintain it for 2~6 hours;

[0021] ④ The shelf temperature is raised to 5°C in 1 to 3 hours, and the shelf temperature is maintained for 1 to 4 hours;

[0022] ⑤ The shelf temperature is raised to 30°C to 40°C within 1 to 5 hours, and the shelf temperature is maintained for 2 to 12 hours; the plug is pressed and the product is taken out of the freeze dryer; after the freeze drying is completed, the plug is pressed and the cap is rolled.

[0023] The method for preparing the lyophilized preparation of thymosin for injection described above, and its further preferred technical scheme is: freeze-drying treatment is performed using the following freeze-drying curve:

[0024] ① Place the qualified products on the freeze dryer shelf, and keep the freeze dryer shelf temperature at 5℃~20℃;

[0025] ②After packing, keep the plate temperature at -40℃ or below, pre-freeze for 30 to 60 minutes until the product temperature is -35 to -45℃, and keep this temperature for 1 to 3 hours;

[0026] ③Raise the shelf temperature quickly to 0℃~5℃ and maintain the shelf temperature for 4~6 hours; when the product temperature reaches -10℃~-7℃, drop the shelf temperature to -5℃ and maintain it for 2~6 hours;

[0027] ④ The shelf temperature is raised to 5°C in 1 to 3 hours, and the shelf temperature is maintained for 1 to 4 hours;

[0028] ⑤ The shelf temperature is raised to 30°C in 1 to 3 hours, and the shelf temperature is maintained for 6 to 12 hours; the plug is pressed and the freeze dryer is taken out; after the freeze drying is completed, the plug is pressed and the cap is rolled.

[0029] The method for preparing the lyophilized preparation of thymosin for injection described above, and its further preferred technical scheme is: freeze-drying treatment is performed using the following freeze-drying curve:

[0030] ① Place the qualified products on the freeze dryer shelf, and keep the freeze dryer shelf temperature at 5℃~20℃;

[0031] ②After packing, keep the plate temperature at -40℃ or below, pre-freeze and cool down for 30 to 60 minutes until the product temperature is -35 to -45℃, and keep this temperature for 1 to 2 hours;

[0032] ③ The shelf temperature rises to 0℃~5℃ in 10min-30min, and the shelf temperature is maintained for 4-6 hours; when the product temperature reaches -10℃~-7℃, the shelf temperature drops to -5℃ and is maintained for 2-4 hours;

[0033] ④ The shelf temperature is raised to 5°C in 1 to 2 hours, and the shelf temperature is maintained for 2 to 4 hours;

[0034] ⑤Raise the shelf temperature to 30°C in 1 to 2 hours and maintain the shelf temperature for 2 to 4 hours

[0035] ⑥ The shelf temperature is raised to 40°C in 0.5 to 1 hour, and the shelf temperature is maintained for 2 to 6 hours, and the plug is pressed and taken out of the freeze dryer; after the freeze drying is completed, the plug is pressed and the cap is rolled.

[0036] The thymosin intermediate solution for injection was prepared and freeze-drying related parameters were measured. The results are shown in the following table:

[0037] Sample name Disintegration temperature Eutectic temperature Eutectic temperature Thymosin solution for injection -7.1℃ -22.20℃ -3.20℃

[0038] According to the measurement parameters, it is usually better to set the primary drying temperature not exceeding the disintegration temperature, and the extreme operation is not to exceed the eutectic temperature. The existing process recommends a primary drying temperature of about -5°C to complete the primary drying. However, as the batch size of this product is enlarged, the end time of the primary drying is greatly extended. On the other hand, due to the long-term lack of further drying, the solid block solidification is achieved. During the product development and amplification process, the technicians found that the freeze-dried products of this product will have a high residual moisture content after batch amplification, which affects the quality of the product during long-term storage. In addition, when the freeze-drying process of this product is enlarged in batches, the freeze-drying time is very long (more than 40 hours), and the product quality cannot be guaranteed. After the technicians compared the batch amplification in detail, they found that the main problem in the primary drying is the imbalance between the energy supply and the energy required for ice crystal sublimation. Because in small batches, in addition to the energy provided by the plate layer, the radiant heat provides enough energy to support the energy demand for sublimation. In large batches, as the volume of the freeze-drying box increases, the radiant heat cannot increase proportionally. Therefore, it is necessary to increase the energy supply of the plate layer, which can significantly shorten the freeze-drying time. Based on this, the inventor creatively raised the plate temperature to a higher temperature (the freezing point of water and above) when the sublimation began. When the product temperature rose to near the disintegration temperature of the product, the plate temperature was lowered to near the disintegration temperature to end the main sublimation and ensure the solid structure of the product. This can significantly shorten the sublimation time, and has better adaptability to batch expansion and consistent product quality.

[0039] Compared with the prior art, the present invention has the following beneficial effects:

[0040] 1. Compared with the prior art, the present invention can significantly shorten the freeze-drying cycle, save electricity, reduce costs, and is suitable for large-scale production. In addition, the product has lower moisture content, better product storage stability, and better product quality.

[0041] 2. The new lyophilized preparation prepared by the method of the present invention has better stability, which is mainly reflected in the lyophilization time, moisture content of the preparation and total impurities inspection items. The lyophilized thymus alfasin for injection produced by this patented method significantly shortens the lyophilization cycle and greatly reduces costs. At the same time, the moisture content is reduced, and the total impurities of the preparation are reduced after the stability test of influencing factors, and are far lower than the import standard JX19990203 for thymus alfasin lyophilized preparations for injection and the provisions in the 2020 edition of the Chinese Pharmacopoeia (the total amount of related substances shall not exceed 4%, and a single impurity shall not exceed 2%). DETAILED DESCRIPTION

[0042] The specific technical solutions of the present invention are further described below to help those skilled in the art further understand the present invention without limiting the rights thereof.

[0043] Example 1, Preparation method of lyophilized preparation of thymosin for injection

[0044] Active ingredients of thymus alkaloid lyophilized preparation for injection: thymus alkaloid 1600mg, mannitol 50g, sodium dihydrogen phosphate, sodium hydrogen phosphate in appropriate amounts; then add water for injection to 1000ml. pH is 6.4. Dispense 1ml / bottle and half-stopper.

[0045] The process also includes preparing the drug of the above formulation into a lyophilized preparation and performing the following treatments:

[0046] ① Place the qualified products on the freeze dryer shelf (0.5m 2 ), the freeze dryer partition temperature was maintained at 20°C;

[0047] ②After packing, set the plate temperature to -40℃, pre-freeze and cool down for 30 minutes, and maintain this temperature for 2 hours;

[0048] ③ The shelf temperature rises to 5°C in 10 minutes and the shelf temperature is maintained for 4 hours; when the product temperature reaches -9°C to -7°C, the shelf temperature drops to -5°C and is maintained for 4 hours;

[0049] ④Raise the shelf temperature to 5°C in 1 hour and maintain the shelf temperature for 2 hours;

[0050] ⑤Raise the shelf temperature to 30°C in 1 hour and maintain the shelf temperature for 2 hours;

[0051] ⑥ The shelf temperature is raised to 40°C in 1 hour and maintained at the shelf temperature for 4 hours; the plug is pressed and the product is taken out of the freeze dryer; after the freeze drying is completed, the plug is pressed and the cap is rolled.

[0052] The total freeze-drying time was 22 hours.

[0053] Example 2 (Comparative Example Patent CN 115998849 A): Preparation method of lyophilized preparation of thymosin for injection

[0054] Active ingredients of thymus alkaloid lyophilized preparation for injection: thymus alkaloid 1600mg, mannitol 50g, sodium dihydrogen phosphate, sodium dihydrogen phosphate in appropriate amounts; then add water for injection to 1000ml. pH is 7.3. Dispense 1ml / bottle, half stoppered.

[0055] The process also includes preparing the drug of the above formulation into a lyophilized preparation and performing the following treatments:

[0056] a: Place the qualified products on the freeze dryer shelf (0.5m for experimental freeze dryer)2 ), the freeze dryer partition temperature is maintained at 20°C; after packing, set the plate temperature to -40°C, pre-freeze and cool down time to 30 minutes, and keep the product temperature below -35°C; and maintain for another 3 hours;

[0057] b: The condenser is cooled to below -50°C, and the vacuum is evacuated to a vacuum degree of ≤0.3 mbar. The temperature rise program is set, and the shelf temperature is raised to -30°C in 2 hours, and the shelf temperature is maintained for 4 hours;

[0058] c: The shelf temperature is raised to -15°C over 2 hours and maintained at the shelf temperature for 10 hours;

[0059] e: The shelf temperature is raised to 15°C in 1 hour;

[0060] f: Raise the shelf temperature to 40°C over 1 hour, maintain for 8 hours, press the stopper, and exit the freeze dryer.

[0061] The total freeze-drying time was 31.5 hours.

[0062] Example 3, Preparation method of lyophilized preparation of thymosin for injection

[0063] Active ingredients of thymus alkaloid lyophilized preparation for injection: thymus alkaloid 1600mg, mannitol 50g, sodium dihydrogen phosphate, sodium hydrogen phosphate in appropriate amounts; then add water for injection to 1000ml. pH is 6.4. Dispense 1ml / bottle and half-stopper.

[0064] The process also includes preparing the drug of the above formulation into a lyophilized preparation and performing the following treatments:

[0065] ① Place the qualified products on the freeze dryer shelf (0.5m 2 ), the freeze dryer partition temperature was maintained at 20°C;

[0066] ②After packing, set the plate temperature to -40℃, pre-freeze and cool down for 30 minutes, and maintain this temperature for 2 hours;

[0067] ③ The shelf temperature rises to 5°C in 30 minutes and is maintained for 4 hours; when the product temperature reaches -9°C to -7°C, the shelf temperature drops to -5°C and is maintained for 3 hours;

[0068] ④Raise the shelf temperature to 5°C in 1 hour and maintain the shelf temperature for 2 hours;

[0069] ⑤ The shelf temperature is raised to 30°C within 2 hours, and the shelf temperature is maintained for 10 hours; the plug is pressed and the product is taken out of the freeze dryer; after the freeze drying is completed, the plug is pressed and the cap is rolled.

[0070] The total freeze-drying time was 25 hours.

[0071] Example 4, Preparation Method of Lyophilized Thymosin for Injection

[0072] Active ingredients of thymus alkaloid lyophilized preparation for injection: thymus alkaloid 56g, mannitol 1750g, sodium dihydrogen phosphate, sodium hydrogen phosphate in appropriate amounts; then add water for injection to 35L. pH is 6.4. Dispense 1ml / bottle and half-stopper.

[0073] The process also includes preparing the drug of the above formulation into a lyophilized preparation and performing the following treatments:

[0074] ① Place the filled qualified products on the freeze dryer shelf (commercial freeze dryer 20m 2 ), the freeze dryer partition temperature was maintained at 20°C;

[0075] ②After packing, set the plate temperature to -40℃, pre-freeze and cool down for 30 minutes, and maintain this temperature for 3 hours;

[0076] ③ The shelf temperature rises to 0°C in 30 minutes and the shelf temperature is maintained for 6 hours; when the product temperature reaches -9°C to -7°C, the shelf temperature drops to -5°C and is maintained for 4 hours;

[0077] ④Raise the shelf temperature to 5°C in 1 hour and maintain the shelf temperature for 2 hours;

[0078] ⑤Raise the shelf temperature to 30°C in 2 hours and maintain the shelf temperature for 2 hours;

[0079] ⑥ The shelf temperature is raised to 40°C in 1 hour and maintained at the shelf temperature for 3 hours; the plug is pressed and the product is taken out of the freeze dryer; after the freeze drying is completed, the plug is pressed and the cap is rolled.

[0080] The total freeze-drying time was 25 hours.

[0081] The following are the stability data of the influencing factors of each embodiment sample made by the inventor for 5 days and 10 days:

[0082] According to Example 1, Example 2, Example 3 and Example 4, samples of thymus alfasin freeze-dried preparations for injection were prepared and tested at 40°C ± 2°C and 75% ± 5% RH, and the related substances and the moisture content of the freeze-dried samples were determined respectively (the detection method was the 2020 edition of the Chinese Pharmacopoeia method, and the standard limit set was the internal standard limit). The results are shown in the following table:

[0083]

[0084] *Note: In the implementation of Patent 2, the moisture content of the freeze-dried sample is greater than the internal set limit when it is out of the box;

[0085] #Note: After 10 days of inspection, the total related substances were greater than the internal set limit, close to but not exceeding the Chinese Pharmacopoeia standard limit (total impurities shall not exceed 4%).

[0086] & Note: The moisture content is the maximum value measured (4 corners and the center of each board layer are taken, and a minimum of 10 samples are tested).

[0087] Stability studies have found that the thymosin freeze-dried preparation for injection prepared by the present invention has better stability than the freeze-drying curve (Example 2) provided in patent document CN115998849. The main advantages are the freeze-drying time, lower product moisture and greater stability at high temperature (better degradation of single impurities and total impurities), and the quality is consistent in batch scale-up, which has significant commercial value.

[0088] The freeze-dried thymosin for injection produced by the patented method significantly shortens the freeze-drying cycle, and the quality of batch commercial scale-up is consistent, which has significant commercial value.

Claims

1. A method for preparing a lyophilized preparation of thymalfasin for injection, characterized in that: The active ingredients of each 1000mL of thymosin freeze-dried preparation for injection are: Thymosin 1000-2200mg, mannitol 20-70g, sodium dihydrogen phosphate, sodium hydrogen phosphate as appropriate; Then add water for injection to 1000ml; pH is 6.8-7.3; packaging, half-stoppering to obtain the lyophilized drug solution; The freeze-drying process was performed using the following freeze-drying curve: ① Place the qualified products on the freeze dryer shelf, and keep the freeze dryer shelf temperature at 5℃~22℃; ②After packing, keep the plate temperature at -40℃ or below, pre-freeze and cool down for 30 to 90 minutes until the product temperature is -35 to -45℃, and keep this temperature for 1 to 4 hours; ③Raise the shelf temperature quickly to 0℃~10℃ and maintain the shelf temperature for 4~6 hours; when the product temperature reaches -10℃~-7℃, drop the shelf temperature to -5℃ and maintain it for 2~6 hours; ④ The shelf temperature is raised to 5°C in 1 to 3 hours, and the shelf temperature is maintained for 1 to 4 hours; ⑤ The shelf temperature is raised to 30°C to 40°C within 1 to 5 hours, and the shelf temperature is maintained for 2 to 12 hours; the plug is pressed and the product is taken out of the freeze dryer; after the freeze drying is completed, the plug is pressed and the cap is rolled.

2. The method for preparing the lyophilized preparation of thymalfasin for injection according to claim 1, characterized in that: The freeze-drying process was performed using the following freeze-drying curve: ① Place the qualified products on the freeze dryer shelf, and keep the freeze dryer shelf temperature at 5℃~20℃; ②After packing, keep the plate temperature at -40℃ or below, pre-freeze for 30 to 60 minutes until the product temperature is -35 to -45℃, and keep this temperature for 1 to 3 hours; ③Raise the shelf temperature quickly to 0℃~5℃ and maintain the shelf temperature for 4~6 hours; when the product temperature reaches -10℃~-7℃, drop the shelf temperature to -5℃ and maintain it for 2~6 hours; ④ The shelf temperature is raised to 5°C in 1 to 3 hours, and the shelf temperature is maintained for 1 to 4 hours; ⑤ The shelf temperature is raised to 30°C in 1 to 3 hours, and the shelf temperature is maintained for 4 to 12 hours; the plug is pressed and the freeze dryer is taken out; after the freeze drying is completed, the plug is pressed and the cap is rolled.

3. The method for preparing the lyophilized preparation of thymalfasin for injection according to claim 1, characterized in that: The freeze-drying process was performed using the following freeze-drying curve: ① Place the qualified products on the freeze dryer shelf, and keep the freeze dryer shelf temperature at 5℃~20℃; ②After packing, keep the plate temperature at -40℃ or below, pre-freeze and cool down for 30 to 60 minutes until the product temperature is -35 to -45℃, and keep this temperature for 1 to 3 hours; ③Raise the shelf temperature quickly to 0℃~5℃ and maintain the shelf temperature for 4~6 hours; when the product temperature reaches -10℃~-7℃, drop the shelf temperature to -5℃ and maintain it for 2~6 hours; ④ The shelf temperature is raised to 5°C in 1 to 3 hours, and the shelf temperature is maintained for 2 to 4 hours; ⑤Raise the shelf temperature to 30°C in 1 to 3 hours and maintain the shelf temperature for 2 to 4 hours ⑥ The shelf temperature is raised to 40°C in 0.5 to 2 hours, and the shelf temperature is maintained for 2 to 6 hours, and the plug is pressed and taken out of the freeze dryer; after the freeze drying is completed, the plug is pressed and the cap is rolled.

4. The method for preparing the lyophilized preparation of thymalfasin for injection according to any one of claims 1 to 3, characterized in that: The preparation method of 1000 mL of thymosin freeze-dried preparation for injection is as follows: Step 1: Put the prescribed amount of mannitol into a liquid preparation tank, add 0.8 kg of water for injection, and stir until completely dissolved; Step 2: Take the prescribed amount of raw material thymus fasin and the rinse solution and put them into the batching tank, stirring until completely dissolved; Step 3, after thymus fasin is completely dissolved, the pH value of the solution is adjusted to 6.4-7.3 with phosphate buffer; each time the phosphate buffer is added, it is stirred for 5-10 minutes and then a sample is taken to test the pH value; Step 4: Add water for injection to the batching tank until the total amount is reached; Step 5: Pass the drug solution through a 0.45 μm pre-filter and a 0.22 μm sterilizing filter to a buffer tank for filling; Step 6: adjust the filling amount according to the content of the intermediate drug solution, fill, half-stopper, and put into the freeze drying box for freeze drying.

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