Freeze-drying process of teriparatide for injection

Abstract

The invention discloses a freeze-drying process of teriparatide for injection, the freeze-drying process comprises the steps of dissolving each component of teriparatide for injection and then carrying out freeze-drying treatment, the freeze-drying treatment comprises pre-freezing, primary drying and secondary drying, an annealing step is added in the pre-freezing stage, gradient heating is adopted in the primary drying stage, and the primary drying time is shortened. On the premise of not changing the original prescription, the freeze-drying process of the teriparatide for injection is improved, the annealing step is added, and the one-time drying time is shortened by adopting gradient heating, so that the purposes of reducing the time cost and saving energy can be achieved, the finally prepared product is attractive in appearance, and the content of related substances and the content of water meet the requirements.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly relates to a freeze-drying process for teriparatide for injection. Background Art

[0002] Osteoporosis is a systemic skeletal disease characterized by low bone density and deterioration of bone structure, resulting in reduced bone strength and thus increased susceptibility to fractures. Approximately 80% of fractures are related to osteoporosis. Since fractures can reduce people's quality of life, such as premature death, disability, and increased economic burden, it is crucial to identify individuals at high and extremely high risk of fractures and provide them with adequate treatment options. Teriparatide is a synthetic 34-peptide, which is the 1-34 amino acid fragment of human parathyroid hormone (PTH). This fragment is the biologically active N-terminal region of the endogenous parathyroid hormone PTH containing 84 amino acids, and can regulate bone metabolism, the reabsorption of calcium and phosphorus by renal tubules, and intestinal calcium absorption. This drug is used to treat osteoporosis with a high fracture risk in postmenopausal women, as well as osteoporosis with a high fracture risk induced by continuous glucocorticoid treatment.

[0003] Currently, there are two dosage forms of teriparatide on the market. One is represented by the original teriparatide injection developed by Eli Lilly and Company, and its indication is the treatment of osteoporosis in postmenopausal women with a high risk of fractures. The gender is restricted, so the population using it is limited. The other is the teriparatide freeze-dried powder injection, and the manufacturers are Shanghai United Cell, Asahi Kasei of Japan, and Shenzhen Salubris. It should be noted that only the teriparatide acetate for injection of Asahi Kasei Pharmaceutical of Japan can be used regardless of gender and is suitable for the treatment of osteoporosis with a high fracture risk, such as low bone density, existing fractures, family history of aging and femoral neck fractures. Therefore, considering the indication, applicable population and market of the product, the present invention uses the teriparatide acetate for injection of Asahi Kasei Pharmaceutical as the original preparation for research and development.

[0004] The teriparatide acetate for injection of the Japanese marketed product is a sterile freeze-dried powder injection. Sucrose is used as the freeze-drying excipient and sodium chloride is used as the stabilizer in the prescription. Currently, there are no patents and literature reports on the freeze-drying process for teriparatide acetate for injection. The performance of the final products obtained by using different freeze-drying processes varies greatly, mainly manifested as a long freeze-drying time, high energy consumption, and phenomena such as shrinkage and re-melting of the appearance of the obtained products. In addition, during secondary drying, the temperature can be increased and the time can be extended to reduce the residual moisture content of the finished product, but it may increase the content of related substances in the finished product, thus affecting the quality attributes of the product. Summary of the Invention

[0005] To address the deficiencies in the prior art, the objective of the present invention is to provide a freeze-drying process for teriparatide for injection. Without changing the original research formulation, this freeze-drying process optimizes the freeze-drying process to solve problems such as the relatively long time in the freeze-drying process of existing teriparatide for injection, the appearance atrophy of the product, the re-melting after standing at room temperature for a period of time, and the increase in related substances caused by the freeze-drying process. The present invention is mainly achieved through freeze-drying technology. Currently, freeze-drying technology has emerged as a booming strategy for producing stable drugs and biological products, which can significantly improve the stability of drugs such as proteins, polypeptides, antibiotics, vaccines, and liposomes, as well as greatly enhance the convenience of transportation. Although freeze-drying is considered a very mild and friendly drying process, it can cause many stresses, leading to chemical and physical instabilities. Adding suitable excipients and optimizing freeze-drying process parameters are crucial for obtaining high-quality freeze-dried products. Freeze-drying process parameters (mainly including freezing temperature and rate, chamber vacuum degree, annealing step) and formulation factors including excipient selection are all key factors for the success of the freeze-drying process. In short, drugs after freeze-drying often have advantages such as high stability, long-term preservation ability, and good rehydration performance, making the application of freeze-drying technology in the pharmaceutical field increasingly widespread. One aspect of the present invention provides a freeze-drying process for teriparatide for injection, including: subjecting the teriparatide acetate pharmaceutical solution to freeze-drying, which includes three processes of pre-freezing, sublimation drying (also known as primary drying), and desorption drying (also known as secondary drying) carried out in sequence. During the pre-freezing process, the substance is transformed into a rigid structure. In the primary drying process, the ice crystals formed by the crystallization of free water in the pre-freezing stage are transformed from a solid state to a gaseous state by sublimation. The product temperature (sublimation interface temperature) throughout the process cannot exceed the collapse temperature or eutectic point temperature of the product. In the secondary drying process, the water in the amorphous phase, that is, adsorbed water and bound water, is removed by desorption.

[0006] The specific technical solution of the present invention is as follows:

[0007] The present invention provides a freeze-drying process for teriparatide for injection, wherein the teriparatide for injection includes teriparatide or its pharmaceutically acceptable salt, sucrose, sodium chloride, and water for injection. After dissolving the components of the teriparatide for injection, the pharmaceutical solution is subjected to freeze-drying treatment, and the freeze-drying process includes the following steps:

[0008] S1. Pre-freezing

[0009] In the pre-freezing stage, first set the shelf temperature to -5°C and maintain it for 1 - 2 hours. This operation can make the temperature distribution of the sample relatively uniform and there is no supercooled liquid for crystallization, providing a good crystallization environment for obtaining crystal particles with uniform size and a good crystal grain skeleton structure during the crystallization process. Then lower the shelf temperature to -45°C ± 5°C and maintain it for 1 - 2 hours. This temperature is lower than the glass transition temperature of the product, aiming to completely freeze the sample to reduce the risks such as product spraying, melting, and collapse during the primary drying process of the pre-frozen sample. After the maintenance ends, raise the shelf temperature to -20 ± 5°C and maintain it for 1 - 2 hours for annealing. The introduction of annealing can increase the crystal proportion, enabling the product temperature in the primary drying stage to be increased, the sublimation rate to be increased, and the primary drying duration to be shortened. After the end, lower the shelf temperature to -45 ± 5°C again and maintain it for 1 - 2 hours. Then prepare to cool down the cold trap and evacuate.

[0010] S2. Primary drying (sublimation drying): The primary drying adopts Method I or Method II;

[0011] Method I: Raise the shelf temperature to -40 ± 5°C, and the vacuum degree in the freezer is 80 - 150 μbar, and maintain it for 54 - 56 hours;

[0012] Method II: To improve the drying degree and shorten the time, preferably, the sublimation drying includes the following steps in sequence: (a) Directly raise the shelf temperature to -40°C ± 5°C, and the vacuum degree in the freezer is 80 - 150 μbar, and maintain it for 10 - 20 hours. (b) Directly raise the shelf temperature to -35°C ± 5°C, and the vacuum degree in the freezer is 80 - 150 μbar, and maintain it for 8 - 10 hours. (c) Directly raise the shelf temperature to -30°C ± 5°C, and the vacuum degree in the freezer is 80 - 150 μbar, and maintain it for 8 - 10 hours.

[0013] S3. Secondary drying (desorption drying)

[0014] Since there is still bound water adsorbed on the product after the sublimation drying is completed, this part of the solvent is removed by desorption drying. Since the removal of bound water is more difficult than that of free water, in order to achieve the removal effect as quickly as possible and ensure the porousness of the sample, evacuate to below 0.05 mbar shown by the capacitance vacuum machine within 10 minutes, and directly raise the shelf temperature to 45°C ± 5°C within 2 hours, with a heating rate of 1 ± 0.5°C / min, and maintain it for 4 - 5 hours, so as to achieve the purpose of removing the bound water adsorbed in the sample as thoroughly as possible.

[0015] Furthermore, the cooling rate for lowering the shelf temperature to -45°C ± 5°C in step S1(2) is 0.3 - 0.4°C / min.

[0016] Further, in step S1(3), the heating rate for raising the shelf temperature to -20 ± 5°C is 0.4 - 0.5°C / min.

[0017] Further, in step S1(4), the cooling rate for lowering the shelf temperature to -45 ± 5°C is 0.2 - 0.5°C / min.

[0018] Further, in step S2, for Method I, the time for raising the shelf temperature is 10 - 20 min, preferably 10 min.

[0019] Further, in step S2, for Method I, the time for raising the shelf temperature is 25 - 35 min, preferably 30 min.

[0020] Further, in step S2, for Method I: Raise the shelf temperature to -40°C, and maintain the vacuum in the freezer at 80 - 150 μbar for 54 - 56 h.

[0021] Further, in step S2, for Method II: Perform the following steps in sequence: First, raise the shelf temperature to -45°C, and maintain the vacuum in the freezer at 80 - 150 μbar for 10 - 12 h; then raise the shelf temperature to -40°C, and maintain the vacuum in the freezer at 80 - 150 μbar for 10 - 12 h; then raise the shelf temperature to -35°C, and maintain the vacuum in the freezer at 80 - 150 μbar for 8 - 10 h; finally, raise the shelf temperature to -30°C, and maintain the vacuum in the freezer at 80 - 150 μbar for 8 - 10 h.

[0022] Further, in step S2, for Method I: Raise the shelf temperature to -40°C, and maintain the vacuum in the freezer at 80 μbar for 55 h, and the time for raising the temperature is 10 min.

[0023] Further, in step S2, for Method II: Perform the following steps in sequence: First, raise the shelf temperature to -45°C, and maintain the vacuum in the freezer at 80 μbar for 10 h; then raise the shelf temperature to -40°C, and maintain the vacuum in the freezer at 80 μbar for 10 h; then raise the shelf temperature to -35°C, and maintain the vacuum in the freezer at 80 μbar for 8 h; finally, raise the shelf temperature to -30°C, and maintain the vacuum in the freezer at 80 μbar for 8 h, and the time for raising the temperature in each stage is 30 min.

[0024] The beneficial effects of the present invention are as follows:

[0025] The freeze-drying process of the present invention solves the problems of the long freeze-drying process time of the existing teriparatide for injection, the increase in the content of related substances in the finished product, and the easy shrinkage, collapse or re-melting of the freeze-dried product. Specifically:

[0026] (1) For the entire freeze-drying process of teriparatide for injection, by adding an annealing step and increasing the temperature gradient during the primary drying process, the sublimation drying time can be shortened to a certain extent, achieving the purpose of reducing time costs and energy consumption, and greatly reducing processing costs.

[0027] (2) During the primary drying process, keep the temperature at a low level (below the collapse temperature of the sample) until all sublimation is completed, ensuring that there is no collapse or shrinkage of the sample during the freeze-drying process, and a freeze-dried product with a complete skeleton can be obtained. The moisture content and related substances in the sample meet the requirements.

[0028] (3) During the secondary drying process, by using a high vacuum degree and increasing the drying temperature, the adsorbed water in the product can be removed to the greatest extent while maintaining a good skeleton structure. The moisture content in the finally prepared freeze-dried preparation can be as low as 2% or less. Description of the Drawings

[0029] Figure 1 It is the appearance diagram of the freeze-dried products of Example 1, Example 2, Example 3 and Example 4. Detailed Embodiments

[0030] To better understand the present invention, the present invention will be further described with reference to the following examples and drawings. The examples are only for explanation and do not limit the present invention in any way. In the examples, all original reagent materials can be obtained commercially. The experimental methods without specific conditions are the conventional methods and conditions well-known in the art, or the conditions recommended by the instrument manufacturer.

[0031] The formulation of teriparatide for injection of the present invention refers to the formulation of the original research drug, and the formulation information is shown in Table 1 below:

[0032] Table 1 Formulation Information of Teriparatide for Injection

[0033] Serial Number Component Function Dosage 1 Teriparatide Acetate Active Ingredient 63.3 μg (calculated as teriparatide) 2 Sucrose Excipient 11.2 mg 3 Sodium Chloride Stability 0.56 mg 4 Water for Injection Solvent Add up to 0.56 g

[0034] The following further illustrates the freeze-drying process of teriparatide for injection of the present invention in combination with examples.

[0035] Example 1

[0036] Formulation Preparation Process: Take 5.6 g of sucrose and 280 mg of sodium chloride and dissolve them in 140 g of injection water. After dissolution, take 34.77 mg of teriparatide acetate and dissolve it. Finally, make up the volume to 280 g with injection water. Filter with a 0.22 μm filter, fill 0.56 mL into each vial, and semi-cork.

[0037] Lyophilization process: The teriparatide acetate solution is lyophilized. The lyophilization includes steps of pre-freezing, primary drying, secondary drying, and taking out of the chamber. The details of the lyophilization process are shown in Table 2 below. Among them, the pre-freezing stage is not only to protect the main properties of the substance unchanged, but also to obtain a reasonable structure of the frozen product to facilitate the sublimation of water, and to completely freeze the product to avoid spraying phenomenon during the sublimation process. Theoretically, the pre-freezing temperature should be lower than the glass transition temperature of the product. Therefore, in this embodiment, the pre-freezing temperature is set at -45°C, which is lower than the glass transition temperature of -38°C. The primary drying temperature is maintained at -40°C throughout the process, which is lower than the collapse temperature of the product of -36°C, and the sublimation time is extended. The appearance diagram of the lyophilized product is as Figure 1 , through the optimization of the lyophilization process, the powder cake presents a perfect cake shape without shrinkage, but the time is too long, with a total duration of 87 hours.

[0038] Table 2 Lyophilization process of Example 1

[0039]

[0040] Example 2

[0041] The prescription information and solution preparation method of Example 2 are the same as those of Example 1. The lyophilization process of Example 2 adds an annealing step. The details of the lyophilization process are shown in Table 3 below. Among them, annealing refers to the process of raising the temperature of the frozen sample above the glass transition temperature of the product and below the eutectic temperature, keeping it for a period of time, and then lowering the temperature to the freezing temperature. Adding an annealing step before sublimation drying mainly has two reasons: First, during the freezing process, especially the rapid freezing process, the crystalline components in the formula often do not have enough time to completely crystallize. However, if this component can provide necessary support for the structure of the lyophilized drug or the protein will be more stable after the component is completely crystallized, it is necessary to make it completely crystallized. When the annealing temperature is higher than the glass transition temperature of the concentrated solution of the formula, it will promote the formation of recrystallization and make the crystalline components and unfrozen water crystallize completely. The annealing duration is related to the formula composition and heating rate. Second, through annealing, the glass transition temperature of the amorphous phase can be increased. In this embodiment, the annealing temperature is -20°C and it is maintained for 2 hours, which shortens the sublimation time of the primary drying to a certain extent. The appearance diagram of the lyophilized product is as Figure 1 , through the optimization of this lyophilization process, the powder cake is complete without shrinkage, the lyophilization duration is shortened, and the water content and related substance content are within the requirements.

[0042] Table 3 Lyophilization process of Example 2

[0043]

[0044] Example 3

[0045] Example 3 The prescription information and liquid preparation method are the same as those in Example 1. Based on adding an annealing step to the lyophilization process of Example 3, the primary drying is carried out with a gradient temperature increase to shorten the drying time. The lyophilization process is shown in Table 4 below. To achieve a faster drying speed, the drug temperature is required to be as high as possible, but it must be lower than the eutectic temperature or collapse temperature to prevent the drug from melting, denaturing, or collapsing. During the sublimation stage, heat is mainly provided by conduction, and the heat of the lyophilized product is mainly obtained from the shelves. Through some experience and literature reports, the temperature decrease of the product caused by the heat taken away during sublimation is approximately 15°C. Therefore, by increasing the temperature in a gradient manner during primary drying to shorten the drying time, the time cost is saved to a great extent. The appearance diagram of the lyophilized product is as shown in Figure 1 。

[0046] Table 4 Lyophilization Process of Example 3

[0047]

[0048]

[0049] Example 4

[0050] Example 4 The prescription information and liquid preparation method are the same as those in Example 1. Based on shortening the drying time by increasing the temperature in a gradient manner during primary drying in Example 4, the annealing temperature and secondary drying temperature are increased. The lyophilization process is shown in Table 5 below. Generally speaking, if the residual moisture of the sample is too high, the drug is likely to be inactivated and its stability deteriorates. When the sublimation process of the sample reaches the end point, all the free water has been removed. To better control the moisture, the shelf temperature of the secondary drying can be set to the highest temperature acceptable for the product. At the same time, the vacuum degree is increased as much as possible under the support of the instrument performance, which can shorten the secondary drying time and is also beneficial to the removal of the residual water in the product. The appearance diagram of the lyophilized product is as shown in Figure 1 , and through the optimization of this lyophilization process in this example, the moisture content is reduced and there is no shrinkage phenomenon in the appearance of the sample.

[0051] Table 5 Lyophilization Process of Example 4

[0052]

[0053]

[0054] The results of the moisture content, impurity IMA content, impurity IMB content, impurity IMK content, impurity IMK content, impurity IMO content, impurity IMF content, impurity IMI content, other single impurity content, and total impurity content of the teriparatide for injection products prepared in Examples 1 to 4 are as shown in Table 6 below:

[0055] Table 6 Detection Results of Related Substances in Examples 1 to 4

[0056]

[0057] From the analysis of the appearance, moisture content and related substances of the freeze-dried products, there are little differences among Examples 1 to 4 and all are acceptable. It shows that adding an annealing step in the pre-freezing stage and drying with a gradient temperature increase in the primary drying will not affect the quality of the product, and to a certain extent, shorten the freeze-drying process duration and save time costs.

Claims

1. A freeze-drying process for teriparatide for injection, characterized in that, The teriparatide for injection comprises teriparatide or a pharmaceutically acceptable salt thereof, sucrose, sodium chloride and water for injection. After dissolving each component of the teriparatide for injection, lyophilization treatment is carried out. The lyophilization process comprises the following steps: S1. Pre-freezing: (1) First, set the shelf temperature to -5°C and maintain for 1 - 2 h; (2) Then lower the shelf temperature to -45°C ± 5°C and maintain for 1 - 2 h; (3) After that, raise the shelf temperature to -20 ± 5°C and maintain for 1 - 2 h; (4) Then lower the shelf temperature to -45 ± 5°C and maintain for 1 - 2 h; (5) After that, prepare the cold trap to cool down and evacuate the air. S2. Primary drying: The primary drying adopts Method I or Method II; Method I: Raise the shelf temperature to -40 ± 5°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 54 - 56 h; Method II: Sequentially carry out the following steps: (a) First, raise the shelf temperature to -40°C ± 5°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 10 - 20 h; (b) Then raise the shelf temperature to -35°C ± 5°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 8 - 10 h; (c) After that, raise the shelf temperature to -30°C ± 5°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 8 - 10 h; S3. Secondary drying: Evacuate the air to below 0.05 mbar within 10 min, raise the shelf temperature to 45°C ± 5°C within 2 h, with a heating rate of 1 ± 0.5°C / min, and maintain for 4 - 5 h.

2. The freeze-drying process according to claim 1, wherein In step S1(2), the cooling rate for lowering the shelf temperature to -45°C ± 5°C is 0.3 - 0.4°C / min.

3. The lyophilization process according to claim 1, wherein In step S1(3), the heating rate for raising the shelf temperature to -20 ± 5°C is 0.4 - 0.5°C / min.

4. The lyophilization process according to claim 1, wherein In step S1(4), the cooling rate for lowering the shelf temperature to -45 ± 5°C is 0.2 - 0.5°C / min.

5. The freeze-drying process according to claim 1, wherein In step S2, the heating time for raising the shelf temperature in Method I is 10 - 20 min.

6. The freeze-drying process according to claim 1, wherein In step S2, the heating time for raising the shelf temperature in Method II is 25 - 35 min.

7. The freeze-drying process according to claim 1, characterized in that, In step S2, Method I: Raise the shelf temperature to -40°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 54 - 56 h.

8. The freeze-drying process according to claim 1, characterized in that, In step S2, Method II: Sequentially carry out the following steps: First, raise the shelf temperature to -45°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 10 - 12 h; Then raise the shelf temperature to -40°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 10 - 12 h; Then raise the shelf temperature to -35°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 8 - 10 h; Finally, raise the shelf temperature to -30°C, and the vacuum degree in the freezing box is 80 - 150 μbar, and maintain for 8 - 10 h.

9. The freeze-drying process according to claim 1, wherein In step S2, Method I: Raise the shelf temperature to -40°C, and the vacuum degree in the freezing box is 80 μbar, and maintain for 55 h, and the heating time is 10 min.

10. The freeze-drying process according to claim 1, characterized in that, Method II in step S2: The following steps are carried out in sequence: First, raise the shelf temperature to -45°C, the vacuum degree in the freezer is 80 μbar, and maintain for 10 h; then raise the shelf temperature to -40°C, the vacuum degree in the freezer is 80 μbar, and maintain for 10 h; then raise the shelf temperature to -35°C, the vacuum degree in the freezer is 80 μbar, and maintain for 8 h; finally, raise the shelf temperature to -30°C, the vacuum degree in the freezer is 80 μbar, and maintain for 8 h. The heating time for each stage is 30 min.

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