Cetrorelix acetate preparation composition for injection and preparation method thereof

Abstract

The invention belongs to the field of pharmaceutical preparations, and particularly relates to a cetrorelix acetate preparation composition for injection and a preparation method thereof.According to the preparation composition, through a specific freeze-drying process including a slow freezing stage, an annealing stage, primary drying and secondary drying, it is effectively guaranteed that the in-batch quality uniformity of a preparation is good, and the long-term stability is excellent; and industrial mass production is facilitated.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparations, specifically relating to an injectable cetrorex acetate formulation and its preparation method. Background Technology

[0002] With the continuous development of modern society, the number of infertile women is gradually increasing, with fallopian tube factors accounting for a large proportion. How to help these women conceive as early as possible is a question that has been explored in the field of assisted reproduction. Cetrorelix acetate is a synthetic decapeptide and a third-generation gonadotropin-releasing hormone (GnRH) antagonist. Cetrorelix acetate for injection was developed by Merck and is a lyophilized powder for injection. It was first launched in Germany in August 1999 under the brand name Cetrotide, with a strength of 0.25mg. In assisted reproductive technology, cetrorelix acetate can prevent premature ovulation in patients undergoing controlled ovarian stimulation, and it has a promising clinical application prospect.

[0003] Cetrorex acetate is poorly soluble in water. When preparing lyophilized formulations, acetic acid is often added to increase the solubility of the active pharmaceutical ingredient. However, as it is a polypeptide compound, it has poor stability and is easily affected by factors such as temperature and acetic acid during preparation and storage, which leads to the generation of impurities such as deamidated cetrorex D-21739 and acetylated cetrorex D-64308, thereby affecting the stability of the formulation.

[0004] CN117883393A discloses a method for preparing cetrorex lyophilized powder injections by pre-freezing, sublimation, and re-drying. The pre-freezing, sublimation, and re-drying are all carried out in two stages. Specifically, the two stages of pre-freezing are: the first stage involves cooling to -10℃ to -18℃ and holding at this temperature for at least 0.5 hours; the second stage involves cooling to -35℃ to -45℃ and holding at this temperature for at least 1 hour. CN112807418A discloses a method for preparing injectable cetrorex by freezing a mixed solution containing mannitol and cetrorex acetate in three stages: the first stage involves cooling to -20℃ to -40℃ and holding at this temperature for 0.1 to 2 hours; the second stage involves heating to -20℃ to -5℃ and holding at this temperature for 0.1 to 2 hours; and the third stage involves cooling to 20℃ to -40℃ and holding at this temperature for 0.1 to 2 hours.

[0005] However, the inventors discovered that the freeze-dried contents of the products prepared according to the freeze-drying process described in the aforementioned patent exhibit significant quality variations between bottles from the same batch, mainly in related substances, moisture, and residual acetic acid. The homogeneity of the formulation needs further improvement. Therefore, it is still necessary to develop a suitable preparation method for cetrorex acetate freeze-dried formulations to overcome the shortcomings of the existing technology. Summary of the Invention

[0006] To address the deficiencies in existing technologies, this invention provides a novel injectable cetrorelix acetate formulation and its preparation method. The injectable cetrorelix acetate formulation, through a specific freeze-drying process involving a slow freezing stage, an annealing stage, a primary drying stage, and a secondary drying stage, effectively ensures good batch-to-batch quality uniformity and excellent long-term stability, which is beneficial for large-scale industrial production.

[0007] This invention provides an injectable cetrorex acetate formulation composition, the composition comprising cetrorex acetate and a sugar alcohol excipient, and prepared by a lyophilization process, wherein the lyophilization process comprises four steps: a slow freezing stage, an annealing stage, a primary drying stage, and a secondary drying stage; the weight ratio of cetrorex to the sugar alcohol excipient in the composition is 1:200-240 based on the weight of cetrorex.

[0008] Furthermore, calculated as a percentage by weight of the formulation, the residual acetic acid content in the injectable cetrorex acetate formulation composition is ≤0.3%.

[0009] In a preferred embodiment of the present invention, the sugar alcohol excipient is selected from one or more combinations of mannitol, glucosyl alcohol, sorbitol, and galactitol, more preferably mannitol.

[0010] The weight ratio of cetrorex and sugar alcohol excipients in the composition is 1:215-225, more preferably 1:219.2.

[0011] The slow freezing stage involves lowering the plate temperature to -35°C to -45°C at a cooling rate of 0.1 to 0.2°C / min and then maintaining the temperature thereafter.

[0012] The annealing stage involves raising the plate temperature to -5℃ to 0℃ at a heating rate of 0.5 to 1℃ / min and holding it at that temperature, and then lowering the plate temperature to -35℃ to -45℃ at a cooling rate of ≤0.2℃ / min and holding it at that temperature.

[0013] The first drying process involves first raising the plate temperature to -15 to -10°C at a heating rate of 0.2 to 0.5°C / min and holding it at that temperature, and then raising it to 0 to 5°C at a rate of 0.1 to 0.5°C / min and holding it at that temperature.

[0014] The secondary drying process involves raising the plate temperature to 25°C–35°C at a heating rate of 0.2–0.8°C / min and then maintaining that temperature.

[0015] Preferably, the residual acetic acid content in the injectable cetrorex acetate formulation composition is ≤0.3% by weight percentage, more preferably ≤0.2%.

[0016] The present invention also provides a method for preparing an injectable cetrorex acetate formulation composition, characterized in that the preparation method is a freeze-drying process, comprising four steps: a slow freezing stage, an annealing stage, a primary drying stage, and a secondary drying stage.

[0017] The slow freezing stage involves lowering the plate temperature to -35°C to -45°C at a cooling rate of 0.1 to 0.2°C / min and then maintaining the temperature thereafter.

[0018] The annealing stage involves raising the plate temperature to -5℃ to 0℃ at a heating rate of 0.5 to 1℃ / min and holding it at that temperature, and then lowering the plate temperature to -35℃ to -45℃ at a cooling rate of ≤0.2℃ / min and holding it at that temperature.

[0019] The first drying process involves first raising the plate temperature to -15 to -10°C at a heating rate of 0.2 to 0.5°C / min and holding it at that temperature, and then raising it to 0 to 5°C at a rate of 0.1 to 0.5°C / min and holding it at that temperature.

[0020] The secondary drying process involves raising the plate temperature to 25°C–35°C at a heating rate of 0.2–0.8°C / min and then maintaining that temperature.

[0021] Preferably, the vacuum degree of the primary drying is controlled at 0.2 to 0.4 mbar, more preferably 0.3 mbar; the vacuum degree of the secondary drying is controlled at 0.1 to 0.2 mbar, more preferably 0.1 mbar.

[0022] Preferably, the cooling rate during the slow freezing stage is 0.15℃ / min, and the holding time is ≥1 hour, more preferably ≥3 hours.

[0023] Preferably, the heating rate during the annealing stage is 0.5–0.8 °C / min, and the holding time is ≥2 hours; the cooling rate is ≤0.15 °C / min, more preferably 0.10–0.13 °C / min, and the holding time is ≥1 hour, more preferably ≥2 hours.

[0024] Preferably, the plate temperature during the annealing stage is first raised to -3 to 0°C, and then lowered to -40°C.

[0025] Preferably, the first drying process involves raising the plate temperature to -10°C at a heating rate of 0.2–0.3°C / min, more preferably at a heating rate of 0.25°C / min, and holding the temperature for ≥12 hours; then raising the temperature to 5°C at a rate of 0.2–0.5°C / min, more preferably at a rate of 0.25°C / min, and holding the temperature for ≥6 hours.

[0026] Preferably, the secondary drying heating rate is 0.2-0.5℃ / min, more preferably 0.4-0.5℃ / min, and the holding time is ≥12 hours.

[0027] Preferably, the plate temperature for the secondary drying is 25-30°C, more preferably 30°C.

[0028] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0029] (1) The present invention adopts a special freeze-drying process. By controlling the temperature and time of the pre-freezing and drying stages, it effectively solves the problem of large quality differences between vials in the existing injectable cetrirexate, reduces the risk of bottle spraying and collapse, and ensures uniform and stable product quality within the batch, which is conducive to industrial mass production.

[0030] (2) The lyophilized contents of cetrirexate prepared by this invention have good morphology, rapid reconstitution, and can be completely dissolved within 30 seconds. At the same time, the content of related substances, acetic acid and water residues are low, and the long-term stability of the preparation is better.

[0031] (3) The freeze-drying process of the present invention has good universal applicability to the preparation of injectable cetrorex acetate formulations. During the freeze-drying process, most of the acetic acid and water in the formulation can be effectively removed, reducing residues and ensuring that the product stability is not affected by the amount of acetic acid used. Detailed Implementation

[0032] The present invention is further illustrated below by way of embodiments, but these embodiments are not intended to limit the invention to the scope of the embodiments described. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

[0033] Unless otherwise stated, the material information for the following embodiments and comparative examples is all commercially available.

[0034] Example 1

[0035] Step 1: Preparation of cetrorex acetate solution

[0036] Table 1. Example 1: Unit Dosage Prescription of Cetrolex Acetate Solution

[0037]

[0038]

[0039] Add approximately 360g of water for injection to a 500ml beaker, then add 12.6g of acetic acid. Accurately weigh 100mg of cetrorelix acetate (calculated as cetrorelix) and add it to the above solution, stirring until completely dissolved. Then add 21.92g of mannitol and stir until completely dissolved. Finally, bring the volume to 400ml with water for injection. Filter using a 0.22-micron filter. Fill vials at 1ml / vial, partially stopper, and place in a pharmaceutical vacuum freeze dryer for vacuum drying.

[0040] Step 2: Prepare cetrorex acetate for injection using the freeze-drying process parameters in the table below.

[0041] Table 2 Parameters for the four stages of the freeze-drying process in Example 1

[0042]

[0043] The freeze-drying process consists of four steps, and the process parameters for each step are shown in Table 1.

[0044] 1) Slow freezing stage: The plate temperature is lowered from room temperature to 5°C and held for 60 min. Then, the plate temperature is lowered to -40°C at a cooling rate of 0.15°C / min and held for 180 min.

[0045] 2) Annealing stage: The plate temperature is raised to 0℃ at a heating rate of 0.67℃ / min and held for 180min. Then, the temperature is lowered to -40℃ again at a cooling rate of 0.13℃ / min and held for 120min.

[0046] 3) First drying: Raise the plate temperature to -10℃ at a heating rate of 0.25℃ / min and hold for 720min, then continue to raise the temperature to 5℃ at a heating rate of 0.25℃ / min and hold for 360min; the vacuum degree is controlled at 0.3mbar.

[0047] 4) Secondary drying: The temperature of the plate is raised to 30℃ at a heating rate of 0.42℃ / min and held for 720min; the vacuum degree is controlled at 0.1mbar.

[0048] Example 2

[0049] Step 1: The preparation of cetrorex acetate solution is the same as in Example 1.

[0050] Step 2: Change the annealing stage plate temperature to -3℃, and prepare the injectable cetrorex acetate formulation according to the freeze-drying process parameters in the table below.

[0051] Table 3 Parameters for the four stages of the freeze-drying process in Example 2

[0052]

[0053] Example 3

[0054] Step 1: Preparation of cetrorex acetate solution, by changing the acetic acid content in the prescription, otherwise the same as in Example 1.

[0055] Table 4. Unit Dosage Prescription of Cetrolex Acetate Solution in Example 3

[0056] Drug components Prescription quantity Percentage (w / v) Cetrilac acetate 0.25 mg (calculated as cetrorex) 0.025％ Mannitol 54.8mg 5.48％ Acetic acid 2mg 0.2％ Water for Injection Add to 1ml 94.295％

[0057] Step 2: Using the same freeze-drying process as in Example 1, prepare cetrorex acetate for injection.

[0058] Comparative Example 1

[0059] Step 1: The preparation of cetrorex acetate solution is the same as in Example 1.

[0060] Step 2: Without annealing, prepare the cetrorex acetate formulation for injection according to the freeze-drying process parameters in the table below.

[0061] Table 5. Parameters for the three stages of the freeze-drying process in Comparative Example 1

[0062]

[0063] Comparative Example 2

[0064] Step 1: The preparation of cetrorex acetate solution is the same as in Example 1.

[0065] Step 2: Change the cooling rate during the pre-freezing stage and prepare the cetrorex acetate formulation for injection according to the freeze-drying process parameters in the table below.

[0066] Table 6. Parameters for the four stages of the freeze-drying process in Comparative Example 2

[0067]

[0068] Comparative Example 3

[0069] Step 1: The preparation of cetrorex acetate solution is the same as in Example 1.

[0070] Step 2: Change the cooling rate during the pre-freezing stage and prepare the cetrorex acetate formulation for injection according to the freeze-drying process parameters in the table below.

[0071] Table 7. Parameters for the four stages of the freeze-drying process in Comparative Example 3

[0072]

[0073] Comparative Example 4

[0074] Step 1: The preparation of cetrorex acetate solution is the same as in Example 1.

[0075] Step 2: Change the temperature of the plate during the secondary drying stage and prepare the cetrorex acetate formulation for injection according to the freeze-drying process parameters in the table below.

[0076] Table 8. Parameters for the four stages of the freeze-drying process in Comparative Example 4

[0077]

[0078] Comparative Example 5

[0079] Step 1: The preparation of cetrorex acetate solution is the same as in Example 1.

[0080] Step 2: Change the heating rate during the drying stage and prepare the cetrorex acetate formulation for injection according to the freeze-drying process parameters in the table below.

[0081] Table 9. Parameters for the four stages of the freeze-drying process in Comparative Example 5

[0082]

[0083] Experimental Example 1

[0084] The contents of related substances, acetic acid, and residual moisture at day 0 were measured at the four corners and the middle position of the freeze-dried plates corresponding to the examples and comparative examples. The test scheme was based on the imported drug registration standard JX20130315 for cetrorex acetate for injection. The test results are shown in the table below.

[0085] Table 10 Data on the content of relevant substances, acetic acid, and residual moisture.

[0086]

[0087]

[0088] According to the data in the table, the products prepared by the freeze-drying process in Examples 1-3 have low content of related substances, acetic acid and moisture residue. The quality difference between freeze-dried cetrirexate formulations located in different positions within the same batch is minimal, and the formulations have good stability and uniformity.

[0089] Comparative Example 1 had no annealing stage, and Comparative Example 2 had a faster pre-freezing rate. The resulting products had significantly higher levels of related substances and residual moisture, with large inter-bottle variations and inconsistent batch quality. Comparative Example 3 kept the overall pre-freezing time essentially unchanged but reduced the cooling rate during the slow freezing stage. The resulting products had smaller inter-bottle variations, but the levels of related substances, acetic acid, and residual moisture were all higher than in Examples 1-3, leading to a decrease in product quality. Comparative Example 4 had a higher secondary drying temperature, resulting in smaller inter-bottle variations, but a significantly higher level of related substances and decreased product stability. Comparative Example 5 had an excessively rapid heating rate during the primary drying stage, greatly increasing the risk of contents spraying out and collapsing, resulting in poor morphology that did not meet product production standards. It also led to increased residual acetic acid and moisture in the product.

[0090] Experimental Example 2

[0091] The formulations prepared by the freeze-drying process of Examples 1 and 2 and Comparative Examples 2 and 3 were tested for the stability of related substances under high temperature (40°C) and accelerated conditions (25°C / 60% RH), respectively. The test results are shown in the table below.

[0092] Table 11 Data on the content of relevant substances

[0093]

[0094] According to the data in the table, the residual acetic acid and related substances in the contents of Examples 1 and 2 after lyophilization were low, indicating good formulation stability. Comparative Examples 2 and 3 had higher residual acetic acid and higher impurity content, especially the D-21739 impurity, which showed a significantly faster growth rate under high temperature conditions, which is detrimental to the long-term stability of the formulation.

Claims

1. A method for preparing an injectable cetrorexate acetate formulation, characterized in that, The preparation method is a freeze-drying process, which includes four steps: slow freezing stage, annealing stage, primary drying and secondary drying; The slow freezing stage involves lowering the plate temperature to -35°C to -45°C at a cooling rate of 0.1 to 0.2°C / min and then maintaining the temperature thereafter. The annealing stage involves raising the plate temperature to -5℃ to 0℃ at a heating rate of 0.5 to 1℃ / min and holding it at that temperature, and then lowering the plate temperature to -35℃ to -45℃ at a cooling rate of ≤0.2℃ / min and holding it at that temperature. The first drying process involves first raising the plate temperature to -15 to -10°C at a heating rate of 0.2 to 0.5°C / min and holding it at that temperature, and then raising it to 0 to 5°C at a rate of 0.1 to 0.5°C / min and holding it at that temperature. The secondary drying process involves raising the plate temperature to 25°C–35°C at a heating rate of 0.2–0.8°C / min and then maintaining that temperature.

2. The method for preparing the cetrorex acetate formulation composition for injection according to claim 1, characterized in that, The residual acetic acid content in the cetrorex acetate formulation composition for injection is ≤0.3% by weight percentage.

3. The method for preparing the cetrorex acetate formulation composition for injection according to claim 2, characterized in that, The residual acetic acid content is ≤0.2%.

4. The method for preparing the cetrorex acetate formulation composition for injection according to claim 1, characterized in that, The vacuum level for the first drying is controlled at 0.2–0.4 mbar; the vacuum level for the second drying is controlled at 0.1–0.2 mbar.

5. The method for preparing the cetrorex acetate formulation composition for injection according to claim 1, characterized in that, The plate temperature for the secondary drying is 25–30°C.

6. The method for preparing the cetrorex acetate formulation composition for injection according to claim 1, characterized in that, The cooling rate during the slow freezing stage is 0.15℃ / min.

7. The method for preparing the cetrorex acetate formulation composition for injection according to claim 1, characterized in that, The heating rate during the annealing stage is 0.5–0.8 °C / min, and the cooling rate is ≤0.15 °C / min.

8. The method for preparing the cetrorex acetate formulation composition for injection according to claim 1, characterized in that, The first drying process involves raising the plate temperature to -10°C at a rate of 0.2–0.3°C / min, and then raising it to 5°C at a rate of 0.2–0.5°C / min.

9. The method for preparing the cetrorex acetate formulation composition for injection according to claim 1, characterized in that, The secondary drying heating rate is 0.2–0.5 °C / min.

10. A cetrorex acetate formulation composition for injection, characterized in that, The composition comprises cetrorex acetate and a sugar alcohol excipient, and is prepared by the preparation method according to any one of claims 1-9, wherein the weight ratio of cetrorex to sugar alcohol excipient in the composition is 1:200-240 based on the weight of cetrorex.

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