Manufacturing method for large-grade boronophenylalanine freeze-dried powder injection and freeze-dried powder injection

Abstract

The present invention provides a method for producing a large-grade boronophenylalanine lyophilized powder for injection, and the lyophilized powder for injection, which belongs to the pharmaceutical technical field. The method for producing a large-grade boronophenylalanine lyophilized powder for injection includes the steps of mixing boronophenylalanine, a polyol, an alkali, and a portion of a solvent to obtain a first mixed solution; adjusting the pH of the first mixed solution with a pH adjuster and adding the remaining solvent to obtain a second mixed solution, where the second mixed solution has a boronophenylalanine concentration of 160-250 mg / mL; and dispensing the second mixed solution and then lyophilizing it, which includes pre-freezing, sublimation drying, and analytical drying. The production method of the present invention enables the production of a highly stable, large-grade boronophenylalanine lyophilized powder for injection. Furthermore, the method of the present invention is characterized by a stable process and a short lyophilization time, significantly reducing lyophilization costs.

Description

[Technical Field]

[0001] The present invention belongs to the field of medicine, and particularly relates to a method for preparing large-scale boronophenylalanine lyophilized powder for injection and the lyophilized powder for injection prepared by the method. [Background technology]

[0002] L-BPA is a boron-containing agent used in boron neutron capture therapy, which involves the production of 10 9 However, because L-BPA has a low boron loading and limited targeting, the dosage is very large, reaching 100-500 mg / kg, which is 30 g per dose for an adult weighing 60 kg. L-BPA formulations are usually L-BPA-sugar alcohol solutions, but because L-BPA-sugar alcohol solutions are temperature sensitive and difficult to store, they are generally freeze-dried.

[0003] Due to limitations on freeze-drying concentration and filling volume, conventional freeze-drying processes are generally limited to products with specifications of less than 1g. Low-specification freeze-dried products have the following disadvantages compared to L-BPA, which requires a single 30g dose. Multiple vials must be used at a time, each one opened and dissolved, resulting in cumbersome operations and increased sterility risks. The increased packaging materials not only increase costs but also place a burden on the environment. However, large-specification freeze-dried formulations have a high solids content, making water sublimation difficult. This generally results in long freeze-drying times and high freeze-drying costs. Furthermore, melting during heating or removal of the ice crystal support structure after drying can cause various morphological collapses, making it difficult to form stabilizers, which can affect quality indicators such as dissolution and product stability. Large-specification freeze-dried formulations are difficult to develop, and commercially available freeze-dried powder formulations rarely exceed 3g specifications.

[0004] Patent CN103100094B discloses a freeze-drying process for L-BPA, but the L-BPA content per vial is only 30 mg, which is insufficient for clinical administration. Patent CN113546048A discloses a method for reducing the osmotic pressure of boron-containing injections, but the osmotic pressure of the solution is reduced by nanofiltration before freeze-drying, which is complicated to operate. In addition, the freeze-drying process for a 3 g / vial specification includes an annealing step, which takes 109 hours, which is time-consuming, energy-intensive, and costly.

[0005] Therefore, in order to produce large-grade boronophenylalanine freeze-dried powder for injection and solve the existing technical problems, it is urgently necessary to provide a method for producing large-grade boronophenylalanine freeze-dried powder for injection. Summary of the Invention

[0006] The object of the present invention is to provide a method for producing large-grade boronophenylalanine freeze-dried powder for injection, which can produce high-stability, large-grade boronophenylalanine freeze-dried powder for injection, and the method of the present invention is characterized by a stable process and a reasonable freeze-drying time.

[0007] To achieve the above objectives, the present invention adopts the following technical solutions:

[0008] In a first aspect of the present invention, Mixing boronophenylalanine, a polyol, an alkali, and a part of a solvent to obtain a first mixed solution; adjusting the pH value of the first mixed solution to 7.5 to 8.5 with a pH adjuster, and adding the remaining solvent to obtain a second mixed solution, wherein the concentration of boronophenylalanine in the second mixed solution is 160 to 250 mg / mL; dispensing the second mixed solution and then freeze-drying, the freeze-drying including pre-freezing, sublimation drying and analytical drying.

[0009] In some embodiments of the present invention, the boronophenylalanine includes at least one of 2-boronophenylalanine, 3-boronophenylalanine, and 4-boronophenylalanine (BPA). For example, the boronophenylalanine may be 4-boronophenylalanine (BPA), such as L-BPA or D-BPA. For example, the boronophenylalanine may be 2-boronophenylalanine and / or 3-boronophenylalanine.

[0010] In some embodiments of the present invention, the concentration of boronophenylalanine in the second mixed solution may be 160 mg / mL, 170 mg / mL, 180 mg / mL, 190 mg / mL, 200 mg / mL, 210 mg / mL, 220 mg / mL, 230 mg / mL, 240 mg / mL, or 250 mg / mL. The concentration of the lyophilization liquid used for lyophilization of boronophenylalanine lyophilized powder formulations is generally low, and the use of a lyophilization liquid exceeding 100 mg / mL has rarely been disclosed in the prior art. The product concentration significantly affects the compactability and resolubility of the lyophilized powder.

[0011] In some embodiments of the present invention, the pre-freezing conditions include a temperature of -60°C to -20°C and a pressure of normal pressure. Preferably, the pre-freezing temperature is -50°C to -40°C. For example, the pre-freezing temperature is -60°C, -59°C, -57°C, -55°C, -52°C, -50°C, -49°C, -48°C, -47°C, -46°C, -45°C, -44°C, -43°C, -42°C, -41°C, -40°C, -38°C, -35°C, -33°C, -30°C, -28°C, -26°C, -24°C, -22°C, or -20°C. The normal pressure for pre-freezing is typically 1 atmosphere.

[0012] In some embodiments of the present invention, the pre-freezing time is 2 to 10 hours. Preferably, the pre-freezing time is 2 to 5 hours. For example, the pre-freezing time may be 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, or 10 hours.

[0013] In some embodiments of the present invention, the pre-freezing is performed by maintaining the temperature at -50°C for 2 to 10 hours. The pre-freezing process of the present invention does not require an annealing step, which shortens the freeze-drying time and reduces the process cost and operational difficulty.

[0014] In an embodiment of the present invention, the pre-freezing step of the manufacturing method requires a short time, which effectively reduces the pre-freezing cost.

[0015] In some embodiments of the present invention, the conditions for the sublimation drying include a temperature of -10°C to -2°C and a pressure of 10 to 20 Pa. For example, the temperature for the sublimation drying may be -10°C, -9°C, -8°C, -7°C, -6°C, -5°C, -4°C, -3°C, or -2°C. For example, the pressure for the sublimation drying is 10 Pa, 11 Pa, 12 Pa, 13 Pa, 14 Pa, 15 Pa, 16 Pa, 17 Pa, 18 Pa, 19 Pa, or 20 Pa. Through the above pre-freezing step, the sublimation drying of the present invention can achieve sublimation drying of a highly concentrated boronophenylalanine solution in a single sublimation, and can effectively remove moisture.

[0016] In some embodiments of the present invention, the sublimation drying time is 24 to 54 hours, for example, 24 hours, 25 hours, 26 hours, 27 hours, 30 hours, 32 hours, 35 hours, 37 hours, 39 hours, 40 hours, 42 hours, 45 hours, 47 hours, 49 hours, 50 hours, 53 hours, or 54 hours.

[0017] In some embodiments of the present invention, the sublimation drying is performed at a temperature of -5°C for 24 to 54 hours. Further, the sublimation drying is performed at a temperature of -5°C and a pressure of 10 to 20 Pa for 24 to 54 hours. The sublimation drying process of the present invention does not require an annealing step, which increases the primary drying temperature, shortens the freeze-drying time, achieves the desired drying effect, and reduces process costs and operational difficulties.

[0018] In the preparation method of the embodiment of the present invention, the high concentration of boronophenylalanine in the mixed solution of the present invention and the processing temperature of the sublimation drying step can be combined to avoid the collapse phenomenon of the freeze-dried product. As can be seen from the above, the sublimation drying temperature of the present invention can not only obtain a freeze-dried powder with a loose appearance, but also improve the efficiency of sublimation drying, reduce the drying time, and further reduce the drying cost.

[0019] In some embodiments of the present invention, the conditions for the analysis drying include a temperature of 25° C. to 35° C. and a pressure of 10 to 20 Pa. For example, the temperature for the analysis drying may be 25° C., 26° C., 27° C., 28° C., 29° C., 30° C., 31° C., 32° C., 33° C., 34° C., or 35° C. For example, the pressure for the analysis drying may be 10 Pa, 11 Pa, 12 Pa, 13 Pa, 14 Pa, 15 Pa, 16 Pa, 17 Pa, 18 Pa, 19 Pa, or 20 Pa.

[0020] In some embodiments of the present invention, the analytical drying time is 5 to 15 hours. Preferably, the analytical drying time is 12 to 14 hours. For example, the analytical drying time may be 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 12.5 hours, 13 hours, 13.5 hours, 14 hours, or 15 hours.

[0021] In the production method of the present invention, the solution can be frozen by the pre-freezing, and this state is advantageous for the later drying operation. The sublimation drying of the present invention positions the water line at the bottom of the bottle, and further analytical drying can produce a freeze-dried powder with a loose, clumpy appearance.

[0022] In some embodiments of the present invention, the mass ratio of the boronophenylalanine to the polyol is 1:1 to 1.3, for example, the mass ratio of the boronophenylalanine to the polyol may be 1:1, 1:1.2, or 1:1.3.

[0023] The polyol is at least one selected from fructose, sorbitol, mannitol, erythritol, maltitol, lactitol, and xylitol.

[0024] In some embodiments of the present invention, the alkali is at least one selected from sodium hydroxide, magnesium hydroxide, potassium hydroxide, and calcium hydroxide.

[0025] In some embodiments of the present invention, the pH adjuster is at least one selected from hydrochloric acid, acetic acid, phosphoric acid, nitric acid, and sulfuric acid.

[0026] In some embodiments of the present invention, the solvent is water for injection.

[0027] In some embodiments of the present invention, the second mixed solution is sterilized before the freeze-drying, and the sterilization treatment includes filtering the second mixed solution through a microfiltration membrane, specifically filtering the second mixed solution through 0.45 micron and 0.22 micron filtration membranes.

[0028] In some embodiments of the present invention, the height of the liquid after dispensing is 15 to 30 mm. Specifically, the height of the liquid in the container after dispensing may be 15 mm, 17 mm, 19 mm, 20 mm, 22 mm, 24 mm, 26 mm, 28 mm, or 30 mm.

[0029] In some embodiments of the present invention, the volume of each bottle after dispensing is 15 to 30 mL, for example, 15 mL, 16 mL, 18 mL, 20 mL, 22 mL, 25 mL, 26 mL, 28 mL, or 30 mL.

[0030] In some embodiments of the present invention, the height of a 10 milliliter fill volume in a container after dispensing is approximately 10 millimeters, meaning that in the following examples, the height of the liquid can be estimated based on the fill volume after dispensing.

[0031] In some embodiments of the present invention, the amount of boronophenylalanine contained in each dispensed bottle is 3 to 7.5 grams, for example, 3 grams, 3.5 grams, 4 grams, 5 grams, 6 grams, 7 grams, or 7.5 grams.

[0032] In a second aspect of the present invention, there is provided a lyophilized powder for injection produced by the method according to the first aspect of the present invention. The lyophilized powder for injection contains boronophenylalanine. Furthermore, the content of boronophenylalanine in each tube of the lyophilized powder for injection is 3 to 7.5 grams.

[0033] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (e.g., in the Examples) can be combined with each other to form new or preferred technical solutions, and due to space limitations, detailed descriptions will be omitted here.

[0034] Regarding pre-freezing, if the product is not frozen beforehand, sublimation drying cannot be carried out, and the pre-freezing process not only protects the main properties of the material, but also gives the product a reasonable structure after freeze-drying to help the water sublimate. Pre-freezing is a very important part of the freeze-drying process, if the temperature is too high, the product will not be able to freeze, which will cause bubbles and eruptions during the subsequent sublimation and may result in product loss, and if the temperature is too low, the freezing time will be longer, especially for large-scale formulations.

[0035] Sublimation drying, also known as the first stage of drying, is the most critical and problem-prone stage of freeze-dried products, and temperature must be strictly controlled. The sublimation drying temperature must be lower than the product's collapse temperature or the maximum allowable temperature. If the sublimation drying temperature is too low, the drying efficiency will be slow, the drying time will be long, and it will be difficult to obtain a loose product in a short time.

[0036] Regarding analytical drying, analytical drying is also called second-stage drying. In the first-stage drying, water is removed in the form of ice crystals, but because the adsorption energy of the ice crystals is high compared to adsorbed water, the water cannot be resolved from the adsorption unless sufficient energy is provided.

[0037] The beneficial effects of the present invention are as follows: The method for producing large-grade boronophenylalanine lyophilized powder for injection of the present invention simplifies the operation, reduces sterility risks and production costs, the content of boronophenylalanine in each bottle of produced lyophilized powder for injection is high, and the number of bottles used is relatively reduced. Furthermore, the method of the present invention has a short lyophilization time and can reduce lyophilization costs.

[0038] The manufacturing method of the present invention is easy to operate, and its freeze-drying process does not have an annealing step, the primary drying temperature is increased, the freeze-drying time is shortened, and the process cost and operation difficulty are reduced.

[0039] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, specific embodiments of the present application will be described in detail with reference to the drawings, to make the technical solutions and other beneficial effects of the present application more apparent. [Brief explanation of the drawings]

[0040] [Figure 1] FIG. 1 is an external view of the freeze-dried powder according to Example 1 of the present invention. [Figure 2] FIG. 1 is an external view of the freeze-dried powder according to Example 2 of the present invention. [Figure 3] FIG. 1 is an external view of the freeze-dried powder according to Example 3 of the present invention. [Figure 4] FIG. 1 is an external view of the freeze-dried powder according to Comparative Example 1. DETAILED DESCRIPTION OF THE INVENTION

[0041] The present invention will be further described below with reference to specific examples. It should be understood that the following description is merely the most preferred embodiment of the present invention and should not be considered to limit the scope of protection of the present invention. With a full understanding of the present invention, experimental methods in the following examples, for which specific conditions are not specified, are generally performed under general conditions or conditions suggested by manufacturers. Those skilled in the art may make modifications that are not essential to the technical solutions of the present invention, and such modifications should be considered to fall within the scope of protection of the present invention. The terms "first," "second," "third," etc. (if present) in the present specification and claims and the above drawings are not necessarily intended to describe a specific order or chronological order, but are intended to distinguish between similar objects. It should be understood that the objects described in this manner may be substituted for each other where appropriate. Furthermore, the terms "comprise" and "have," and any variations thereof, are intended to be non-exclusive.

[0042] Example 1 This example provides a method for preparing large-scale boronophenylalanine lyophilized powder injection, including the following steps: Prepare the following formulation: [Table 1]

[0043] The manufacturing process is as follows: S1, the formulation of BPA and fructose were weighed and added to 300 mL of water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4: Water was added to make the volume 600 mL to obtain a solution containing 160 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 25 mL of intermediate solution. S7: freeze-dried, the BPA content in the resulting product was 4 g / bottle.

[0044] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying, where pre-freezing is performed at a temperature of -50°C for 5 hours, sublimation drying is performed at a temperature of -5°C and a pressure of 10 Pa for 35 hours, and analytical drying is performed at a temperature of 30°C and a pressure of 10 Pa for 12 hours.

[0045] Example 2 This example provides a method for preparing large-scale boronophenylalanine lyophilized powder injection, including the following steps: Prepare the following formulation: [Table 2]

[0046] The manufacturing process is as follows: S1, the formulation of BPA and fructose were weighed and added to 300 mL of water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4: Water was added to make the volume 600 mL to obtain a solution containing 200 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 25 mL of intermediate solution. S7: freeze-dried, and the BPA content in the resulting product was 5 g / bottle.

[0047] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying, where pre-freezing is performed at a temperature of -50°C for 5 hours, sublimation drying is performed at a temperature of -5°C and a pressure of 10 Pa for 35 hours, and analytical drying is performed at a temperature of 30°C and a pressure of 10 Pa for 12 hours.

[0048] Example 3 This example provides a method for preparing large-scale boronophenylalanine lyophilized powder injection, including the following steps: Prepare the following formulation: [Table 3]

[0049] The manufacturing process is as follows: S1, the formulation of BPA and fructose were weighed and added to 300 mL of water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4: Water was added to make the volume 600 mL to obtain a solution containing 200 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 30 mL of intermediate solution. S7: freeze-dried, the BPA content in the resulting product was 6 g / bottle.

[0050] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying, where pre-freezing is performed at a temperature of -50°C for 5 hours, sublimation drying is performed at a temperature of -5°C and a pressure of 10 Pa for 35 hours, and analytical drying is performed at a temperature of 30°C and a pressure of 10 Pa for 12 hours.

[0051] Example 4 Example 4 provides a method for preparing large-scale boronophenylalanine lyophilized powder injection, which differs from Example 1 in the following respects: As shown in Table 3, the BPA concentration of Example 4 is 250 mg / mL, and the BPA content of each dispensed bottle is 6.25 grams. Other conditions are similar to those of Example 1.

[0052] Example 5 Example 5 provides a manufacturing method for large-scale boronophenylalanine lyophilized powder injection, which differs from Example 1 in the following respects: As shown in Table 3, the filling volume of each bottle in Example 5 is 15 mL, and the BPA content of each bottle after dispensing is 2.4 grams. Other conditions are similar to those in Example 1.

[0053] Examples 6 to 9 Examples 6 to 9 provide methods for preparing large-scale boronophenylalanine lyophilized powder injections, which differ from Example 1 in the following respects: As shown in Table 3, the pre-freezing temperatures in Examples 6 to 9 are -60°C, -40°C, -30°C, and -20°C, respectively; the other conditions are almost the same as those in Example 1.

[0054] Examples 10-11 Examples 10 and 11 provide methods for preparing large-scale boronophenylalanine lyophilized powder injections, which differ from Example 1 in the following respects: As shown in Table 3, the pre-freezing durations of Examples 10 and 11 are 2 hours and 10 hours, respectively; other conditions are almost the same as those of Example 1.

[0055] Examples 12-13 Examples 12 and 13 provide a method for producing large-scale boronophenylalanine freeze-dried powder injections, which differ from Example 1 in the following respects: As shown in Table 3, the sublimation drying temperatures in Examples 12 and 13 are -10°C and -2°C, respectively. The other conditions are almost the same as those in Example 1.

[0056] Examples 14 to 16 Examples 14 to 16 provide methods for preparing large-scale boronophenylalanine lyophilized powder injections, which differ from Example 1 in the following respects: As shown in Table 3, the duration of sublimation drying in Examples 14 to 16 is 24 hours, 54 hours, and 30 hours, respectively; other conditions are similar to those in Example 1.

[0057] Examples 17-18 Examples 17 and 18 provide a method for preparing large-scale boronophenylalanine lyophilized powder injections, which differ from Example 1 in the following respects: As shown in Table 3, the analytical drying temperatures in Examples 17 and 18 are 25°C and 35°C, respectively; other conditions are almost the same as those in Example 1.

[0058] Examples 19-20 Examples 19 and 20 provide a method for preparing large-scale boronophenylalanine lyophilized powder injections, which differ from Example 1 in the following respects: As shown in Table 3, the analytical drying durations of Examples 19 and 20 are 5 hours and 14 hours, respectively; other conditions are almost the same as those of Example 1.

[0059] Comparative Example 1 Comparative Example 1 provides a preparation method for boronophenylalanine lyophilized powder injection, which includes the following steps: S1, BPA and fructose were weighed and added to water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4, water was added to the volume to obtain a solution containing 160 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 25 mL of intermediate solution. S7: freeze-dried, the BPA content in the resulting product was 4 g / bottle.

[0060] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying, where pre-freezing is performed at a temperature of -50°C for 5 hours, sublimation drying is performed at a temperature of -20°C and a pressure of 10 Pa for 35 hours, and analytical drying is performed at a temperature of 30°C and a pressure of 10 Pa for 12 hours.

[0061] Comparative Example 2 Comparative Example 2 provides a preparation method for boronophenylalanine lyophilized powder injection, which includes the following steps: S1, BPA and fructose were weighed and added to water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4, water was added to the solution to obtain a solution containing 50 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 80 mL of intermediate solution. S7: freeze-dried, the BPA content in the resulting product was 4 g / bottle.

[0062] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying, where pre-freezing is performed at a temperature of -50°C for 5 hours, sublimation drying is performed at a temperature of -5°C and a pressure of 10 Pa for 35 hours, and analytical drying is performed at a temperature of 30°C and a pressure of 10 Pa for 12 hours.

[0063] Comparative Example 3 Comparative Example 3 provides a preparation method for boronophenylalanine lyophilized powder injection, which includes the following steps: S1, BPA and fructose were weighed and added to water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4, water was added to the solution to obtain a solution containing 80 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 50 mL of intermediate solution. S7: freeze-dried, the BPA content in the resulting product was 4 g / bottle.

[0064] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying, where pre-freezing is performed at a temperature of -50°C for 5 hours, sublimation drying is performed at a temperature of -5°C and a pressure of 10 Pa for 35 hours, and analytical drying is performed at a temperature of 30°C and a pressure of 10 Pa for 12 hours.

[0065] Comparative Example 4 Comparative Example 4 provides a preparation method for boronophenylalanine lyophilized powder injection, which includes the following steps: S1, BPA and fructose were weighed and added to water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4, water was added to the volume to obtain a solution containing 160 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 25 mL of intermediate solution. S7: freeze-dried, the BPA content in the resulting product was 4 g / bottle.

[0066] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying, where pre-freezing is performed at a temperature of -10°C for 5 hours, sublimation drying is performed at a temperature of -5°C and a pressure of 10 Pa for 35 hours, and analytical drying is performed at a temperature of 30°C and a pressure of 10 Pa for 12 hours.

[0067] Comparative Example 5 Comparative Example 5 provides a method for preparing boronophenylalanine lyophilized powder injection, which includes the following steps: S1, BPA and fructose were weighed and added to water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4: Water was added to the solution to obtain a solution containing 40 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 25 mL of intermediate solution. S7: freeze-dried, and the BPA content in the resulting product was 1 g / bottle.

[0068] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying, where pre-freezing is performed at a temperature of -50°C for 5 hours, sublimation drying is performed at a temperature of -5°C and a pressure of 10 Pa for 35 hours, and analytical drying is performed at a temperature of 30°C and a pressure of 10 Pa for 12 hours.

[0069] Comparative Example 6 Comparative Example 6 provides a method for preparing boronophenylalanine lyophilized powder injection, which includes the following steps: S1, BPA and fructose were weighed and added to water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4, water was added to the volume to obtain a solution containing 160 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 25 mL of intermediate solution. S7: freeze-dried, the BPA content in the resulting product was 4 g / bottle.

[0070] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying. Pre-freezing includes steps 1, 2, and 3, where step 1 is held at -50°C for 5 hours, step 2 is held at -35°C for 3 hours, and step 3 is held at -55°C for 4 hours. As shown in Table 1, sublimation drying is held at -5°C and 10 Pa pressure for 28 hours, and analytical drying is held at 30°C and 10 Pa pressure for 12 hours.

[0071] Comparative Example 7 Comparative Example 7 provides a method for preparing boronophenylalanine lyophilized powder injection, which includes the following steps: S1, BPA and fructose were weighed and added to water for injection. S2, an appropriate amount of sodium hydroxide was added and stirred until the solution became clear. S3, hydrochloric acid was added to adjust the pH to 7.5 to 8.5. S4, water was added to the volume to obtain a solution containing 160 mg / mL BPA. S5, the solution was sterilized and filtered through 0.45 micron and 0.22 micron filter membranes to obtain an intermediate solution. S6, Filled and filled the vessel with 25 mL of intermediate solution. S7: freeze-dried, the BPA content in the resulting product was 4 g / bottle.

[0072] In this example, freeze-drying includes pre-freezing, sublimation drying, and analytical drying. Pre-freezing includes steps 1, 2, and 3, where step 1 is holding at -50°C for 5 hours, step 2 is holding at -35°C for 3 hours, and step 3 is holding at -55°C for 4 hours. As shown in Table 1, sublimation drying is holding at -5°C and 10 Pa pressure for 35 hours, and analytical drying is holding at 30°C and 10 Pa pressure for 12 hours.

[0073] Test Example 1 In this test example, the freeze-dried powders of Example 1 of the present invention and Comparative Examples 6 to 7 were studied, and some parameters of Example 1 and Comparative Examples 6 to 7 are shown in Table 1. The appearance of the freeze-dried powder obtained in each group was observed, and the freeze-dried powder was subjected to detection of moisture and redissolution, and the structure was recorded, and the details are shown in Table 2.

[0074] [Table 4]

[0075] [Table 5]

[0076] From Tables 1 and 2, it can be seen that the freeze-dried powder prepared in Example 1 of the present invention has the appearance of white loose lumps, a reconstitution time of 60 seconds, a moisture content of 0.57%, and the advantages of short freeze-drying time and complete drying, and the preparation method of the present invention can achieve excellent freeze-drying effect.

[0077] The pre-freezing in Comparative Examples 6 and 7 employed an annealing process, and a comparison with Example 1 of the present invention reveals the following: In Comparative Example 7, after the annealing process was employed, the total freeze-drying time had to be lengthened to achieve the technical effect of freeze-drying, and the freeze-dried powder produced was the same as in Example 1 of the present invention, both of which were white and loose lumps; in Comparative Example 6, after the annealing process was employed, the total freeze-drying time of Example 1 of the present invention was used, but complete freeze-drying could not be achieved, and the resulting freeze-dried powder was not completely dry, and the reconstitution time was long at 180 seconds, with a high moisture content of 4.87%.

[0078] Although an annealing process was used in Comparative Example 7, the appearance, reconstitution time, and water content of the freeze-dried powder produced were almost the same as those of the freeze-dried powder produced in Example 1 of the present invention. Therefore, it can be seen that the annealing process of Comparative Example 7 does not have a better effect and has drawbacks such as being time-consuming and expensive.

[0079] From the above, it can be seen that the manufacturing method of the present invention not only can produce large-scale freeze-dried powder products with excellent performance, but also can significantly shorten the total freeze-drying time of large-scale BPA, improve production efficiency, and save production costs, making it worthy of widespread application.

[0080] Test Example 2 In this test example, performance studies were carried out on the freeze-dried powders produced in Examples 1 to 20 of the present invention and Comparative Examples 1 to 5, respectively, and some parameters for Examples 1 to 20 and Comparative Examples 1 to 5 are shown in Table 3. The appearance of the freeze-dried powders produced in each example was observed, and moisture and redissolution were detected for the freeze-dried powders, and their structures were recorded. The details are shown in Table 4.

[0081] [Table 6] JPEG2026508416000007.jpg56170

[0082] [Table 7]

[0083] According to the descriptions in Tables 3 and 4, As can be seen from Examples 1 to 5, the manufacturing method of the present invention has excellent freeze-drying effect for high concentrations of BPA, and within the concentration loading range of the present invention, good technical effects can be achieved with all of the above process parameters. Specifically, see Figures 1, 2, and 3, where Figure 1 is an external view of the freeze-dried powder manufactured in Example 1, Figure 2 is an external view of the freeze-dried powder manufactured in Example 2, and Figure 3 is an external view of the freeze-dried powder manufactured in Example 3, all of which are white loose lumps. It is clear that the manufacturing process of the present invention can produce white loose lumps of freeze-dried powder, i.e., high-quality BPA injection, and has a short total freeze-drying time, resulting in significant technical effects.

[0084] According to Examples 1, 12, and 13, high-concentration BPA can be effectively freeze-dried at the high sublimation drying temperature of -10°C to -2°C of the present invention, and the appearance, moisture content, and reconstitution time of the freeze-dried powder all meet the application needs. Furthermore, this temperature can improve the efficiency of sublimation drying, reduce the sublimation drying time, and further reduce costs. Referring to Examples 12 and 13, any sublimation temperature within the range of the present invention can achieve good technical effects, and the higher the temperature, the higher the drying efficiency and lower the moisture content in the same time.

[0085] Compared to Examples 1, 12, and 13 of the present invention, Comparative Example 1 had a lower sublimation temperature and lower drying efficiency. Comparative Example 1 was unable to completely sublimate within the same time as Example 1, did not reach the sublimation endpoint, and the bottom of the final product was not completely dry, was pale yellow, had a high moisture content of 4.87%, and ultimately was not completely redissolved even after 180 seconds. Specifically, see Figures 1 and 4. Figure 1 is an external view of the freeze-dried powder produced in Example 1, which appeared as white loose clumps. Figure 4 is an external view of the freeze-dried powder produced in Comparative Example 1, which appeared as a pale yellow bottom that was not completely dry.

[0086] In addition to the drying effect, the manufacturing method of the present invention improves the efficiency of sublimation drying, further shortens the overall freeze-drying time, and saves costs. Generally, the sublimation drying temperature is low, because the sublimation interface temperature of the product must be lower than the collapse temperature; otherwise, the freeze-dried product will collapse and the drying effect will be poor. Thus, the time required for complete sublimation drying of the product is generally relatively long, resulting in high costs. Therefore, the manufacturing method of the present invention has significant technical advantages.

[0087] In Comparative Example 2, the filling volume was 80 mL and the liquid height was approximately 80 mm; in Comparative Example 3, the filling volume was 50 mL and the liquid height was approximately 50 mm. The higher the filling height, the more difficult it was to dry; in Comparative Example 2, one-half was not completely dried and the color was light yellow; in Comparative Example 3, one-third was not completely dried and the color was light yellow. The lyophilized powders produced in Comparative Examples 2 and 3 had high final moisture contents, as high as 8.78%, and could not be completely redissolved even after 180 seconds. Comparing Example 1 with Comparative Examples 2 and 3, it can be seen that the manufacturing method of the present application can produce lyophilized powder injections with a high BPA content, and the performance of the lyophilized powders is excellent, meeting the application requirements.

[0088] In Examples 1, 6, and 7, the pre-freezing temperature was -60°C to -40°C, and the freeze-dried powder produced had a loose, lumpy appearance, low water content, and short re-dissolution time. In Examples 8 and 9, the freeze-dried powder produced had a low water content and short re-dissolution time, but a small amount of erupting occurred. This shows that in the method for producing large-grade BPA of the present invention, the higher the pre-freezing temperature, the worse the freezing effect, and therefore a small amount of erupting may occur during subsequent sublimation.

[0089] In Comparative Example 4, the pre-freezing temperature was -10°C, which was too high, and the high-concentration BPA product could not be completely frozen, and the subsequent sublimation would cause bubbles and relatively serious eruptions, and the re-dissolution time would be long. From the above, it can be seen that in the freeze-drying process of high-concentration BPA, exceeding the pre-freezing temperature of the present invention would result in obvious product loss, and it would take a long time to completely dissolve the drug on the bottle wall during re-dissolution, about 180 seconds.

[0090] In Comparative Example 5, the concentration of the freeze-dried solution was 40 mg / mL. The freeze-dried powder produced by the same freeze-drying process as in Example 1 of the present invention had a collapsed appearance, was unable to form a complete block structure, and the freeze-dried powder had a long redissolution time and a high water content. It is clear that the freeze-dried solution of Comparative Example 5, when combined with the freeze-drying conditions of the Examples of the present invention under the conditions of low concentration and high loading, causes the phenomenon of product collapse, which further affects the freeze-drying effect. As can be imagined, the present invention can not only reduce the collapse of the freeze-dried solution by combining a high-concentration freeze-drying solution with the freeze-drying process of the present invention, but also increase the drying efficiency and reduce the freeze-drying cost.

[0091] According to Examples 10 and 11, the pre-freezing time within the process range of the present invention can achieve a good freezing effect, and the overall pre-freezing time of the present invention is short, which can effectively shorten the total freeze-drying time and save costs.

[0092] According to Examples 14 to 16, the sublimation time within the process range of the present invention can all achieve good drying results. At the same temperature, the time of the present invention can all achieve good drying results, and the longer the time, the better the drying effect at the same temperature and the lower the moisture content, but at the same time the cost also increases. As can be imagined, in addition to pre-freezing, the present invention combines the sublimation drying temperature and holding time of the present invention, all of which have excellent drying effects and further reduce freeze-drying costs.

[0093] According to Examples 17 to 20, good drying results can be achieved under the analytical temperature and analytical time conditions within the process range of the present invention. As can be imagined, within the scope of the present invention, the higher the analytical drying temperature, the higher the drying efficiency and the lower the moisture content in the same time, and the longer the analytical drying time, the higher the drying effect and the lower the moisture content in the same temperature, but at the same time, the cost increases. Because boronophenylalanine is unstable to heat, if the analytical drying temperature exceeds the tolerance temperature of boronophenylalanine, boronophenylalanine will react with polyols, for example, causing a Maillard reaction with fructose, resulting in a yellow appearance and further affecting the quality of the freeze-dried powder.

[0094] In the embodiment of the present invention, by combining the freeze-drying process of the present invention with a high concentration freeze-drying solution, the overall freeze-drying time can be effectively shortened and the drying efficiency can be improved, thereby improving the shortcomings of the prior art.

[0095] In the examples of the present invention, the following method can be used to detect the freeze-dried powder. (1) Appearance: Inspection method: The product was visually inspected under natural light.

[0096] (2) Re-melting time: Test method: Take one bottle of the test sample, pour in an appropriate amount of purified water until the BPA concentration reaches 100 mg / ml, start timing, shake, and stop timing when the contents are completely dissolved, and record the time it takes for the contents to redissolve.

[0097] (3) Moisture: Reagents and reagents: Phosphorus pentoxide (analytical reagent) Equipment and tools: vacuum oven, electronic balance (1:10,000) A weighing bottle was taken and dried under reduced pressure at 40°C using phosphorus pentoxide as a desiccant until it reached a constant weight, designated as M1. Two bottles were taken and crushed by striking the bottle with a blunt object. The contents were removed and quickly crushed, approximately 1g of the product was weighed accurately, and the weight was designated as M2. The bottle was placed in a weighing bottle with a constant weight, dried under reduced pressure at 40°C for 24 hours, cooled in a desiccator, and then weighed again. The weight was designated as M3. The weight loss was calculated using the following formula. Two parallel tests were conducted, and the average value was calculated.

[0098]

number

[0099]

number

[0100] Test Example 3 In this test example, the stability of the freeze-dried powder prepared in Example 1 of the present invention was investigated, including the resolubility, pH, related substances, and content of the freeze-dried powder. Specifically, two batches of samples were prepared according to the method of Example 1, and the content and related substances were measured after standing at 25°C for 0 and 3 months, respectively. The detailed results are shown in Tables 5 and 6.

[0101] The following test method can be used in the present invention. (1) Properties: Inspection method: Visual inspection. Specific test procedure: The product was taken and observed under natural light.

[0102] (2) pH: Testing method: pH value measurement method (Chinese Pharmacopoeia 2015 Edition, Four Parts General Provisions 0631).

[0103] (3) Related substances: HPLC method (Chinese Pharmacopoeia 2015 Edition, Four Parts General Rules 0512).

[0104] (4) Content: HPLC method (Chinese Pharmacopoeia 2015, Part 4 General Rules 0512). The content in this test example was measured as the BPA content.

[0105] [Table 8]

[0106] [Table 9]

[0107] As can be seen from Tables 5 and 6, the properties and pH of the freeze-dried powder are stable after storage at 25°C for 3 months, the content of related impurities is very low, and the content of the active ingredient BPA is stable, meeting the requirements for use. As can be seen from the above, the two batches of samples produced in Example 1 of the present invention have good stability after storage at 25°C for 3 months. This shows that the large-scale freeze-dried powder produced by the production method of the present invention has excellent stability and can meet the requirements for use.

[0108] As described above, the preparation method of the present invention can produce a large-grade boronophenylalanine freeze-dried powder for injection, and the prepared large-grade boronophenylalanine freeze-dried powder has a loose appearance, a low water content, meets the requirements for use and storage, has an excellent freeze-drying effect, and the reconstitution time and stability of the freeze-dried powder are both excellent, so as to meet the requirements for use.In addition, the preparation method of the present invention can significantly shorten the overall freeze-drying time of the large-grade boronophenylalanine, further improve production efficiency, and save production costs, which has great economic value.

[0109] The above are only preferred embodiments of the present invention, and it should be pointed out that those skilled in the art can make several improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered as part of the protection scope of the present invention.

Claims

1. Mixing boronophenylalanine, a polyol, an alkali, and a portion of a solvent to obtain a first mixed solution; adjusting the pH value of the first mixed solution to 7.5-8.5 with a pH adjuster, and adding the remaining solvent to obtain a second mixed solution, wherein the concentration of boronophenylalanine in the second mixed solution is 160-250 mg / mL; a step of dispensing the second mixed solution and then freeze-drying, wherein the freeze-drying includes steps of pre-freezing, sublimation drying and analytical drying.

2. The method according to claim 1, wherein the pre-freezing conditions include a temperature of −60 to −20° C. and atmospheric pressure.

3. 2. The method according to claim 1, wherein the conditions for the sublimation drying include a temperature of -10°C to -2°C and a pressure of 10 to 20 Pa.

4. The manufacturing method according to claim 1, wherein the conditions for the analytical drying include a temperature of 25°C to 35°C and a pressure of 10 to 20 Pa.

5. The method according to any one of claims 1 to 4, wherein the pre-freezing time is 2 to 10 hours.

6. 6. The method according to claim 5, wherein the pre-freezing is performed by maintaining the product at a temperature of -50°C for 2 to 10 hours.

7. The method according to any one of claims 1 to 4, wherein the sublimation drying time is 24 to 54 hours.

8. 8. The method according to claim 7, wherein the sublimation drying is carried out at a temperature of −5° C. for 24 to 54 hours.

9. The manufacturing method according to any one of claims 1 to 4, wherein the analytical drying time is 5 to 15 hours.

10. The mass ratio of the boronophenylalanine to the polyol is 1:1 to 1.

3.

2. The method according to claim 1, wherein the polyol is at least one selected from the group consisting of fructose, sorbitol, mannitol, erythritol, maltitol, lactitol, and xylitol.

11. the alkali is at least one selected from sodium hydroxide, magnesium hydroxide, potassium hydroxide, and calcium hydroxide; and / or the pH adjuster is at least one selected from hydrochloric acid, acetic acid, phosphoric acid, nitric acid, and sulfuric acid; and / or The method according to claim 1, wherein the solvent is water for injection.

12. The method according to claim 1 , wherein the second mixed solution is sterilized before the freeze-drying, and the sterilization treatment includes filtering the second mixed solution through a microfiltration membrane for sterilization.

13. The method according to claim 1, wherein the volume of each bottle after dispensing is 15 to 30 mL.

14. The manufacturing method according to claim 1, wherein the height of the liquid after dispensing is 15 to 30 mm.

15. The method according to claim 1, wherein the amount of boronophenylalanine contained in each bottle after dispensing is 3 to 7.5 grams.

16. A freeze-dried powder injection, produced by the production method according to any one of claims 1 to 15.

17. The freeze-dried powder for injection according to claim 16, wherein the content of boronophenylalanine in each tube of the freeze-dried powder for injection is 3 to 7.5 g.

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