Gabexate mesylate freeze-dried preparation and preparation method thereof

Abstract

The invention provides a gabexate mesylate freeze-dried preparation and a preparation method thereof, and relates to the technical field of medicine preparation. According to the gabexate mesylate preparation method provided by the invention, the pre-freezing process is improved, and large ice crystals are formed in the pre-freezing stage through the annealing process, so that water can be quickly discharged and a large amount of water is removed in the heating process of one-time drying, melting and collapse are avoided, and the appearance is kept complete. In addition, according to the method, quick cooling and freezing are carried out in the later stage of primary drying, so that water formed by melting ice which is not easy to dry in large ice crystals during primary drying is cooled and frozen into ice again, the volume is expanded, the large ice crystals are cracked into small ice crystals in the secondary drying process, and therefore a water flowing channel is formed, water is further removed, and the drying efficiency is improved. Therefore, the water removal rate is high, the product quality is more stable, and the clinical application of the gabexate mesylate freeze-dried preparation is effectively promoted.

Description

Technical Field

[0001] The present invention relates to the technical field of drug preparation, in particular to a lyophilized gabexate mesylate preparation and a preparation method thereof. Background Art

[0002] Gabexate is a non-peptide protein inhibitor that can inhibit the activity of proteases such as trypsin, kinase, plasmin, and thrombin, thereby preventing the pathological and physiological changes caused by these enzymes. In animal experimental acute pancreatitis, it can inhibit activated trypsin and reduce pancreatic damage. At the same time, the increase in serum amylase, lipase activity and urea nitrogen is also significantly improved. It is used to treat acute mild (edema type) pancreatitis and can also be used as an adjuvant treatment for acute hemorrhagic necrotizing pancreatitis. Gabexate mesylate was approved by the Japanese Pharmaceutical Special Committee in 1977 and was produced and sold by Ono Pharmaceutical Industry Co., Ltd. in Japan in 1978 under the trade name FOY.

[0003] Gabexate mesylate has a lyophilized preparation, and the preparation of lyophilized preparations in the current prior art requires the addition of lyophilizing aids in the prescription. Chinese patent CN1911208A discloses a gabexate mesylate injection and a preparation method thereof, and excipients such as 1,2-propylene glycol, sodium chloride, sodium bisulfite, polyethylene glycol, etc. are added to the prescription. Chinese patent CN106038497A discloses a pharmaceutical composition, preparation and preparation method of gabexate mesylate, and the prescription includes sugar alcohol substances such as mannitol, sorbitol, glucose, dextran, etc. and pH regulators such as sulfurous acid, phosphoric acid, nitrous acid or acetic acid. Chinese patent CN102526017A discloses a gabexate mesylate pharmaceutical composition and a preparation method of its lyophilized powder injection, and vitamin C, glycine and dextran are added to the prescription. These solutions have complex prescriptions and high costs, and the addition of excipients increases the quality risk of the injection.

[0004] In addition, the inventors of the present application also found during the research process that, since the raw material of gabexate mesylate is relatively fine and easily soluble in water, the skeleton structure formed during the freeze-drying process is relatively dense, which makes it difficult for water to escape, thereby causing a remelting phenomenon during the heating process. Therefore, the gabexate mesylate product is not easy to form during the freeze-drying process. Summary of the invention

[0005] One of the purposes of the present invention is to provide a method for preparing a lyophilized preparation of gabexate mesylate in view of the above-mentioned problems. The method improves the lyophilization process and obtains a lyophilized preparation of gabexate mesylate with complete appearance and stable quality without adding any lyophilization excipients.

[0006] Another object of the present invention is to provide a lyophilized preparation of gabexate mesylate, which is prepared by the above-mentioned preparation method.

[0007] The technical solution adopted by the present invention is as follows:

[0008] A method for preparing a lyophilized gabexate mesylate preparation comprises adding injection water to dissolve the gabexate mesylate into a solution, filtering the solution to obtain a drug solution, and freeze-drying the drug solution to obtain the lyophilized gabexate mesylate preparation; wherein the freeze-drying comprises:

[0009] Pre-freezing: Cool the liquid medicine to below -25℃ to freeze it, raise the temperature of the frozen solid to -2~-18℃ and anneal for more than 30min, cool the powder cake to below -25℃ again and keep it for 60-180min;

[0010] Primary drying: heat up to 10-30℃ at a rate of 0.1-0.5℃ / min, keep it for no less than 200min, then quickly cool it down to below -10℃ and freeze it, and keep it for 60-120min;

[0011] Secondary drying: Slowly return the temperature to 30-60℃ for secondary drying and maintain for no less than 120 minutes.

[0012] In a preferred embodiment of the present invention, during the pre-freezing process, the drug solution is cooled to -30 to -40°C and then annealed.

[0013] In a preferred embodiment of the present invention, the annealing temperature is -5 to -10°C and the time is 60 to 240 minutes.

[0014] In a preferred embodiment of the present invention, after annealing, the powder cake is cooled again to -30 to -40°C.

[0015] In a preferred embodiment of the present invention, during the primary drying process, the heating rate is 0.2-0.3°C / min.

[0016] In a preferred embodiment of the present invention, the primary drying is heated to 15-20°C.

[0017] In a preferred embodiment of the present invention, the temperature is lowered to -45 to -30°C after primary drying, and the cooling rate is 1.5 to 3°C / min.

[0018] In a preferred embodiment of the present invention, during the secondary drying process, the heating rate of the temperature recovery is 0.2-1°C / min.

[0019] In a preferred embodiment of the present invention, the drug solution is prepared by the following method: a prescribed amount of gabexate mesylate is added to water for injection to prepare a gabexate mesylate solution with a concentration of 2% to 25% W / V, and the gabexate mesylate solution is filtered through a 0.22 micron filter membrane to obtain the drug solution.

[0020] A lyophilized preparation of gabexate mesylate is prepared by any of the above-mentioned preparation methods.

[0021] The beneficial effects of the present invention are:

[0022] The preparation method of gabexate mesylate provided by the present invention improves the pre-freezing process, forms large ice crystals in the pre-freezing stage through an annealing process, and allows the water to quickly drain during the heating process of primary drying, removes a large amount of water, avoids melting and collapse, and keeps the appearance intact. In addition, the method also rapidly cools and freezes in the late stage of primary drying, so that the water melted from the ice that is not easy to dry in the large ice crystals during primary drying is cooled and frozen into ice again, expands in volume, and cracks into small ice crystals during the secondary drying process, thereby forming a channel for draining water, further removing water, thereby removing high water content, and making the product quality more stable, effectively promoting the clinical application of the lyophilized preparation of gabexate mesylate. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 These are photos of the lyophilized gabexate mesylate preparation samples prepared in Examples 1-4 and Comparative Example 3. The samples all appear as complete white powder cakes.

[0024] Figure 2 This is a photo of the lyophilized preparation sample of gabexate mesylate prepared in Comparative Example 1. The sample is completely melted and cannot be formed.

[0025] Figure 3 This is a photo of the lyophilized preparation sample of gabexate mesylate prepared in Comparative Example 2. Part of the sample collapsed and melted.

[0026] Figure 4 This is a cross-sectional electron microscope scanning image of the sample after freeze-drying in Comparative Example 3, showing that the sample is large ice crystals.

[0027] Figure 5 This is a cross-sectional electron microscope scanning image of the sample after freeze-drying in Example 1, showing that the sample is small ice crystals. DETAILED DESCRIPTION

[0028] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention.

[0029] Example 1

[0030] This embodiment provides a method for preparing gabexate mesylate, which specifically comprises the following steps:

[0031] Preparation: Dissolve the prescribed amount of gabexate mesylate in 80% of the prepared amount of water for injection, stir until completely dissolved, make up to volume, filter through a 0.22 micron filter membrane, fill into a vial, and half-stopper.

[0032] Pre-freezing: Cool to -40°C for 180 minutes, maintain for 120 minutes to freeze, raise the temperature of the frozen powder cake to -7°C for 30 minutes to anneal for 180 minutes, cool the powder cake to -40°C for 60 minutes again, maintain for 120 minutes to freeze.

[0033] Primary drying: 240 minutes to 20°C, maintained for 300 minutes, vacuum degree 0.1 mbar. Then 30 minutes to -40°C to freeze the sample again, maintained for 120 minutes, vacuum degree unchanged.

[0034] Secondary drying: heat to 40°C in 240 min, maintain for 300 min, vacuum degree is 0 mbar.

[0035] Example 2

[0036] This embodiment provides a method for preparing gabexate mesylate, which specifically comprises the following steps:

[0037] Preparation: Dissolve the prescribed amount of gabexate mesylate in 80% of the prepared amount of water for injection, stir until completely dissolved, make up to volume, filter through a 0.22 micron filter membrane, fill into a vial, and half-stopper.

[0038] Pre-freezing: Cool to -35°C for 180 minutes, maintain for 120 minutes to freeze, raise the temperature of the frozen powder cake to -5°C for 30 minutes to anneal for 120 minutes, cool the powder cake to -35°C for 60 minutes again, maintain for 120 minutes to freeze.

[0039] Primary drying: 200 minutes to 15°C, maintained for 240 minutes, vacuum degree 0.1 mbar. Then 25 minutes to -35°C to freeze the sample again, maintained for 120 minutes, vacuum degree unchanged.

[0040] Secondary drying: temperature increased to 30°C in 240 min, maintained for 300 min, vacuum degree 0 mbar.

[0041] Example 3

[0042] This embodiment provides a method for preparing gabexate mesylate, which specifically comprises the following steps:

[0043] Preparation: Dissolve the prescribed amount of gabexate mesylate in 80% of the prepared amount of water for injection, stir until completely dissolved, make up to volume, filter through a 0.22 micron filter membrane, fill into a vial, and half-stopper.

[0044] Pre-freezing: Cool to -30°C for 180 minutes, maintain for 120 minutes to freeze, raise the temperature of the frozen powder cake to -10°C for 30 minutes and anneal for 240 minutes, cool the powder cake to -30°C for 60 minutes again, maintain for 120 minutes to freeze.

[0045] Primary drying: 250 minutes to 20°C, maintained for 300 minutes, vacuum degree 0.1 mbar. Then 30 minutes to -30°C to freeze the sample again, maintained for 120 minutes, vacuum degree unchanged.

[0046] Secondary drying: heat to 50°C in 200 min, maintain for 300 min, vacuum degree is 0 mbar.

[0047] Example 4

[0048] This embodiment provides a method for preparing gabexate mesylate, which specifically comprises the following steps:

[0049] Preparation: Dissolve the prescribed amount of gabexate mesylate in 80% of the prepared amount of water for injection, stir until completely dissolved, make up to volume, filter through a 0.22 micron filter membrane, fill into a vial, and half-stopper.

[0050] Pre-freezing: Cool to -40°C for 90 minutes, maintain for 120 minutes to freeze, raise the temperature of the frozen powder cake to -8°C for 30 minutes and anneal for 180 minutes, cool the powder cake to -40°C for 60 minutes again, maintain for 120 minutes to freeze.

[0051] Primary drying: 200 minutes to 20°C, maintained for 360 minutes, vacuum degree 0.1 mbar. Then 20 minutes to -40°C to freeze the sample again, maintained for 120 minutes, vacuum degree unchanged.

[0052] Secondary drying: heat to 60°C in 240 minutes, maintain for 300 minutes, and the vacuum degree is 0 mbar.

[0053] Comparative Example 1

[0054] This comparative example provides a method for preparing gabexate mesylate, which specifically comprises the following steps:

[0055] Preparation: Dissolve the prescribed amount of gabexate mesylate in 80% of the prepared amount of water for injection, stir until completely dissolved, make up to volume, filter through a 0.22 micron filter membrane, fill into a vial, and half-stopper.

[0056] Pre-freezing: Cool to -40°C for 180 minutes and maintain freezing for 120 minutes.

[0057] Primary drying: 120 minutes to 0℃, maintained for 120 minutes, vacuum degree is 0.1mbar. Then 120 minutes to 10℃, maintained for 120 minutes, and 120 minutes to 20℃, maintained for 120 minutes, vacuum degree is maintained at 0.1mbar.

[0058] Secondary drying: heat to 40°C in 60 minutes, maintain for 420 minutes, vacuum degree is 0 mbar.

[0059] Comparative Example 2

[0060] This comparative example provides a method for preparing gabexate mesylate, which specifically comprises the following steps:

[0061] Preparation: Dissolve the prescribed amount of gabexate mesylate in 80% of the prepared amount of water for injection, stir until completely dissolved, make up to volume, the pH value should be between 4.0 and 5.5, filter through a 0.22 micron filter membrane, fill in a vial, and half-stopper.

[0062] Pre-freezing: Cool to -40°C for 180 minutes, maintain for 120 minutes to freeze, raise the temperature of the frozen powder cake to -7°C for 30 minutes to anneal for 180 minutes, cool the powder cake to -40°C for 60 minutes again, maintain for 120 minutes to freeze.

[0063] Primary drying: heat to 0°C over 360 min, maintain for 420 min, vacuum degree 0.1 mbar.

[0064] Secondary drying: temperature increased to 15°C in 300 min, maintained for 420 min, vacuum degree 0.1 mbar.

[0065] Comparative Example 3

[0066] This comparative example provides a method for preparing gabexate mesylate, which specifically comprises the following steps:

[0067] Preparation: Dissolve the prescribed amount of gabexate mesylate in 80% of the prepared amount of water for injection, stir until completely dissolved, make up to volume, the pH value should be between 4.0 and 5.5, filter through a 0.22 micron filter membrane, fill in a vial, and half-stopper.

[0068] Pre-freezing: Cool to -40°C for 180 minutes, maintain for 120 minutes to freeze, raise the temperature of the frozen powder cake to -7°C for 30 minutes to anneal for 180 minutes, cool the powder cake to -40°C for 60 minutes again, maintain for 120 minutes to freeze.

[0069] Primary drying: heat to 20°C in 240 minutes, maintain for 300 minutes, vacuum degree is 0.1 mbar.

[0070] Secondary drying: heat to 40°C in 240 min, maintain for 300 min, vacuum degree is 0 mbar.

[0071] Comparative Example 4

[0072] This comparative example provides a method for preparing gabexate mesylate, which specifically comprises the following steps:

[0073] Preparation: Dissolve the prescribed amount of gabexate mesylate in 80% of the prepared amount of water for injection, stir until completely dissolved, make up to volume, the pH value should be between 4.0 and 5.5, filter through a 0.22 micron filter membrane, fill in a vial, and half-stopper.

[0074] Prefreezing: cool down to -45℃ at a rate of 1℃ / min, freeze for 30min, then heat up to -8℃ at a rate of 0.7℃ / min, and keep warm for 120min.

[0075] Primary drying: Raise the temperature to 5°C and keep warm for 6 hours under a vacuum of 15Pa.

[0076] Secondary drying: heat to 30°C at a rate of 1°C / min and keep warm for 1h under vacuum of 15Pa.

[0077] The appearance of the lyophilized preparations of gabexate mesylate prepared in each embodiment and each comparative example is compared, and the results are shown in Tables 1 and Figure 1-3 As shown. The results show that the appearance of the samples obtained in Examples 1-4 is a complete white powder cake, without melting or collapse. The sample obtained in Comparative Example 1 is completely melted and can hardly be formed. Although the sample obtained in Comparative Example 2 can be formed, it all shows melting and collapse. The appearance of the sample in Comparative Example 3 is also a relatively complete white powder cake, without melting or collapse. The appearance of some samples in Comparative Example 4 is similar to that of the samples in Examples 1-4, which are white freeze-dried blocks, and some samples melt and collapse. According to the above results, it can be seen that without annealing, the sample melts during the first drying process and cannot be formed, such as Comparative Example 1. Comparative Example 2 adds annealing on the basis of Comparative Example 1, and the sample can be formed, but because the gradient heating rate of the first drying is too slow, the water flow is also relatively slow, resulting in partial melting and collapse of the sample. On the basis of Comparative Example 2, Examples 1-4 and Comparative Example 3 increase the heating rate of the first drying, and the appearance of the sample is significantly improved. The inventors analyzed that this is because the crystals are rearranged and combined during the annealing process, which increases the proportion of large ice crystals in the powder cake, causing the collapse temperature and eutectic point temperature to move up, allowing water to drain quickly during a single drying process, removing a large amount of water, and making the sample less likely to melt or collapse. In addition, the inventors also analyzed that in Comparative Example 4, due to the low temperature during the single heating, the removal of water was partially affected, resulting in the melting and collapse of some samples.

[0078] Table 1 Comparison of freeze-dried sample appearance

[0079] category Appearance Example 1 Complete white powder Example 2 Complete white powder Example 3 Complete white powder Example 4 Complete white powder Comparative Example 1 The sample is completely melted Comparative Example 2 The sample melts and collapses Comparative Example 3 Complete white powder Comparative Example 4 White freeze-dried mass (part of the sample melted and collapsed)

[0080] The lyophilized preparations of gabexate mesylate prepared in Examples 1-4 and Comparison 3-4 and the reference preparations were tested according to Part II of the 2020 edition of the Chinese Pharmacopoeia, and the product appearance was compared (the appearance of the samples obtained in Comparative Examples 1-2 was incomplete, so no subsequent samples were retained and tested, and Comparative Example 4 selected samples with complete appearance without melting or collapse). Among them, the manufacturer of the reference preparation is Maruishi Pharmaceutical Co., Ltd., with a batch number of 312421 and a specification of 100 mg. The test and comparison results are shown in Table 2.

[0081] Table 2 Detection and comparison results of different freeze-dried samples

[0082]

[0083]

[0084] According to the results in Table 2, the samples in Examples 1-4, the samples in Comparative Example 3 and the samples with intact appearance in Comparative Example 4 did not change significantly in appearance after stability sintering (60°C-10 days), but after inspection, the ethyl paraben content in the samples in Comparative Examples 3-4 was significantly increased. The inventors measured the microstructures of different samples, and the results are as follows: Figure 4 and Figure 5 As shown. Among them, Figure 4 The cross-sectional electron microscopy image of the sample after freeze-drying in Comparative Example 3 (the cross-sectional electron microscopy image of the sample in Comparative Example 4 is the same as Figure 4 Very similar), Figure 5 The cross-sectional electron microscope scanning diagram of the sample after freeze-drying in Example 1 (the cross-sectional electron microscope scanning diagrams of the samples in Examples 2-4 are similar to Figure 5 same), Figure 4 The ice crystals are large in size, which can increase the water flow rate during the primary drying process. However, large ice crystals are easy to wrap moisture and are difficult to remove, resulting in incomplete water flow during the secondary drying process. According to the inventor's analysis, this is also one of the reasons why the sample in Comparative Example 4 melted and collapsed. Figure 5 In the drying process, ice crystals are broken to form small pieces of ice crystals, which improves the water removal efficiency during the secondary drying process, and the water removal rate is increased, which is conducive to maintaining the integrity of the appearance. In addition, the surface layer of large ice crystals is too dense, and the water in the ice crystals is not easy to detect in the drying loss detection method. Therefore, although the water content detection values ​​of the samples in Comparative Examples 3 and 4 are not excessively high, ethyl paraben increases significantly during the stability sampling process.

[0085] In summary, the present invention improves the freeze-drying process of Gabexate mesylate, that is, on the one hand, large ice crystals are formed in the pre-freezing stage through an annealing process, so that water can be quickly drained during the temperature rise process of primary drying, a large amount of water is removed, melting and collapse are avoided, and the appearance is kept intact. On the other hand, rapid cooling and freezing are performed in the later stage of primary drying, so that the water melted from the ice that is not dried and removed in the large ice crystals during primary drying is cooled and frozen into ice again, the volume expands, and it is cracked into small ice crystals during the secondary drying process, thereby forming a channel for draining water, further removing water, so that the water removal is high, the product quality is more stable, and the clinical application of the lyophilized preparation of Gabexate mesylate is effectively promoted.

Claims

1. A method for preparing a lyophilized preparation of gabexate mesylate, characterized in that: Add injection water to dissolve gabexate mesylate into a solution, filter the solution to obtain a drug solution, and freeze-dry the drug solution to obtain a gabexate mesylate lyophilized preparation; wherein the freeze-drying comprises: Pre-freezing: Cool the liquid medicine to below -25℃ to freeze it, raise the temperature of the frozen solid to -2~-18℃ and anneal for more than 30min, cool the powder cake to below -25℃ again and keep it for 60-180min; Primary drying: heat up to 10-30℃ at a rate of 0.1-0.5℃ / min, keep it for no less than 200min, then quickly cool it down to below -10℃ and freeze it, and keep it for 60-120min; Secondary drying: Slowly return the temperature to 30-60℃ for secondary drying and maintain for no less than 120 minutes.

2. The preparation method according to claim 1, characterized in that: During the pre-freezing process, the drug solution is cooled to -30 to -40°C before annealing.

3. The preparation method according to claim 2, characterized in that: The annealing temperature is -5 to -10°C and the time is 60 to 240 minutes.

4. The preparation method according to claim 3, characterized in that: After annealing, the powder cake is cooled again to -30 to -40°C.

5. The preparation method according to claim 1, characterized in that: During the primary drying process, the heating rate is 0.2-0.3℃ / min.

6. The preparation method according to claim 5, characterized in that: Raise the temperature to 15-20℃.

7. The preparation method according to claim 6, characterized in that: Cool down to -45~-30℃, with a cooling rate of 1.5~3℃ / min.

8. The preparation method according to claim 1, characterized in that: During the secondary drying process, the heating rate of the temperature recovery is 0.2 to 1°C / min.

9. The preparation method according to any one of claims 1 to 8, characterized in that: The drug solution is prepared by the following method: a prescribed amount of gabexate mesylate is added to injection water to prepare a gabexate mesylate solution with a concentration of 2% to 25% W / V; the gabexate mesylate solution is filtered through a 0.22 micron filter membrane to obtain the drug solution.

10. A lyophilized preparation of gabexate mesylate, characterized in that: The preparation method is prepared by the preparation method described in any one of claims 1 to 9.

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