Drying process of botulinum toxin preparations

Abstract

FIELD OF THEINVENTION The present invention relates to a method for drying botulinum toxin under reduced pressure. FIELD OF THEINVENTION The present invention relates to a method for producing a botulinum toxin dried cake under reduced pressure, and the drying time can be significantly shortened while maintaining the efficacy of the botulinum toxin.

Description

Technical Field

[0001] The present invention relates to the drying process of toxin preparations, and specifically to a method for manufacturing a botulinum toxin dry cake by a vacuum drying method.

Background Art

[0002] Botulinum toxin (BTX) is a neurotoxin produced by the anaerobic bacterium Clostridium botulinum, and there are a total of seven types (types A to G). Currently, two types, botulinum A (BTX-A) and B (BTX-B), are purified and used medically.

[0003] Botulinum toxin plays a role in blocking muscle contraction signal transmission and relaxing muscles by suppressing the release of the neurotransmitter acetylcholine. That is, it blocks the release of the neurotransmitter acetylcholine secreted at the presynaptic terminal at the neuromuscular junction, inducing nerve paralysis. There is a difference in that a filler is a medical device that fills a substance at a site where the volume of the skin is insufficient, while a botulinum toxin preparation is a pharmaceutical containing a component that prevents the release of neurotransmitters that cause muscle contraction and reduces muscle use.

[0004] Botulinum toxin is mainly used to suppress or remove wrinkles between the eyebrows and around the eyes, and is also used in the treatment of upper limb rigidity after stroke, eyelid spasm, clubfoot deformity, etc., and its indications are gradually expanding.

[0005] Commercially available botulinum toxin products can be supplied in solution form with excipients including sodium chloride and human serum albumin, or as a solid (dried cake) after a drying process. Most botulinum toxin manufacturers produce products after a freeze-drying process. Examples include Meditoxin®, Xeomin®, and Dysport®.

[0006] On the other hand, while many conventional freeze-drying methods for the drying process of botulinum toxin are publicly known (e.g., KR10-2012-0112248 A), these methods require a freezing process and then remove water through a sublimation process, which takes a considerable amount of time (approximately 18 to 48 hours). In particular, the freezing process, which is an essential step in freeze-drying, can cause ice nuclei to form or imbalances in excipient concentration that occur partially during freezing, leading to a decrease in activity through damage to the protein structure.

[0007] While this freeze-drying process is generally known as an advanced form of drying for desiccants of protein-based pharmaceuticals, it is unsuitable for botulinum toxin preparations that use minute amounts of protein.

[0008] Therefore, there is a need to develop a drying process suitable for the characteristics of botulinum toxin preparations. Furthermore, given the characteristics of the freeze-drying process, a method is needed to efficiently utilize the long process time required for primary and secondary drying. [Overview of the project] [Problems that the invention aims to solve]

[0009] The inventors diligently conducted research to develop an optimized vacuum drying method for botulinum toxin that maintains effectiveness and stability. As a result, they discovered that by efficiently adjusting parameters related to the drying process, such as pressure and (shelf) temperature, it is possible to protect the botulinum toxin, which is a protein, from external stimuli that occur during the process and significantly shorten the drying time, thereby completing the present invention.

[0010] Therefore, the object of the present invention is to provide a method for producing botulinum toxin dried cake. [Means for solving the problem]

[0011] The inventors diligently conducted research to develop an optimized vacuum drying method for botulinum toxin that maintains effectiveness and stability. As a result, they discovered that by efficiently adjusting parameters related to the drying process, such as pressure and (shelf) temperature, it is possible to protect the botulinum toxin, which is a protein, from external stimuli that occur during the process and significantly shorten the drying time.

[0012] This invention relates to a method for producing dried botulinum toxin cake.

[0013] The present invention will now be described in more detail.

[0014] According to one aspect of the present invention, the present invention provides a method for producing a dried botulinum toxin cake, comprising the step of drying botulinum toxin under reduced pressure conditions of 1,500 to 60,000 mTorr and a temperature of 3°C to 25°C.

[0015] In this specification, the term "botulinum toxin (BTX)" refers to the toxin obtained by purification from the bacterium Clostridium botulinum. Botulinum toxin produced by the bacterium Clostridium botulinum has a size of approximately 150 kDa. Botulinum toxin complexes, which are complex substances in which botulinum toxin is bound to one or more non-toxic proteins, have a size of approximately 300 kDa to 900 kDa.

[0016] The aforementioned botulinum toxin can be used for a variety of medical purposes, not only to suppress or improve frown lines and wrinkles around the eyes.

[0017] In this specification, the term "cake" refers to the dried substance on the bottom of a vial-type desiccant form and can be used to distinguish it from powder-type dosage forms.

[0018] In one embodiment of the present invention, the botulinum toxin-producing bacterial strain of the present invention is Clostridium botulinum. More specifically, the Clostridium botulinum Type A (ATCC 19397) strain may be used, but is not limited thereto. Botulinum toxin type A is approved by the U.S. Food and Drug Administration (FDA) for essential blepharospasm, strabismus, and hemifacial spasm in patients 12 years of age and older, and for the treatment of cervical dystonia, frown lines, and hyperhidrosis.

[0019] In another embodiment of the present invention, the botulinum toxin of the present invention may be produced using a variety of conventionally known processes, and is not particularly limited.

[0020] To give a specific example, it may be a culture medium of a botulinum toxin-producing bacterial strain that has undergone one or more precipitation, filtering, redissolution, and purification processes.

[0021] To give a more specific example, the botulinum toxin of the present invention may be a culture solution of a botulinum toxin-producing strain prepared by purifying a botulinum toxin complex from a culture of Clostridium botulinum (C. botulinum) cultured in a specific medium that would be obvious to those skilled in the art, through a series of acid precipitates, into a crystalline complex composed of an active high molecular weight toxin protein and associated hemagglutinin protein, and then dissolving the purified crystalline complex in a solution containing saline and a stabilizer.

[0022] The culture conditions for the botulinum toxin-producing bacterial strain culture can be appropriately adjusted according to the common technical knowledge of those skilled in the art, depending on the culture environment.

[0023] In yet another embodiment of the present invention, the concentration of the botulinum toxin of the present invention may be 250 U / mL to 5,000 U / mL.

[0024] In a specifically preferred embodiment, the concentration of the botulinum toxin is 250 U / mL to 5,000 U / mL, 300 U / mL to 5,000 U / mL, 400 U / mL to 5,000 U / mL, 500 U / mL to 5,000 U / mL, 600 U / mL to 5,000 U / mL, 700 U / mL to 5,000 U / mL, 800 U / mL to 5,000 U / mL, 900 U / mL to 5,000 U / mL, 1,000 U / mL to 5,000 U / mL, 1,000 U / mL to 4,500 U / mL, 1,000 U / mL to 4,000 U / mL, and 1,000 U / mL to 3 The concentrations may be 500 U / mL, 1,000 U / mL to 3,000 U / mL, 1,000 U / mL to 2,500 U / mL, 1,000 U / mL to 2,000 U / mL, 1,000 U / mL to 1,500 U / mL, 1,000 U / mL to 1,200 U / mL, 800 U / mL to 1,200 U / mL, 800 U / mL to 1,100 U / mL, 800 U / mL to 1,000 U / mL, 800 U / mL to 900 U / mL, 900 U / mL to 1,200 U / mL, 1,000 U / mL to 1,200 U / mL, or 1,100 U / mL to 1,200 U / mL.

[0025] In yet another embodiment of the present invention, the method for producing a dried botulinum toxin cake of the present invention may be one in which the above-mentioned botulinum toxin is dried under reduced pressure at a pressure of 1,500 to less than 60,000 mTorr.

[0026] In a specifically preferred embodiment, the vacuum drying of the present invention is carried out at a pressure of 1,500 to 60,000 mTorr, 1,500 to 55,000 mTorr, 1,500 to 50,000 mTorr, 1,500 to 45,000 mTorr, 1,500 to 40,000 mTorr, 1,500 to 35,000 mTorr, 1,500 to 30,000 mTorr, 1,500 to 25,000 mTorr, 1,500 to 20,000 mTorr, 1,500 to 15,000 mTorr, 1,500 to 14,000 mTorr, 1,500 to 13,000 mTorr, 1,500 to 12,000 mTorr, 1,500 to 11,000 mTorr, 1,500 to 10,000 mTorr, 1,500 to 9,000 mTorr, 1,500 to 8,000 mTorr, 1,500 to 7,000 mTorr, 1,500 to 6,000 mTorr, 1,500 to 5,000 mTorr, 1,500 to 4,000 mTorr, 1,500 to 3,500 mTorr, 1,500 to 3,000 mTorr, 1,500 to 2,500 mTorr, 1,500 to 2,000 mTorr, 2,500 to 3,000 mTorr, 2,500 to 3,500 mTorr, 2,500 to 4,000 mTorr, 2,500 to 4,500 mTorr, 2,500 to 5,000 mTorr, 2,500 to 6,000 mTorr, 2,500 to 7,000 mTorr, 2,500 to 8,000 mTorr, 2,500 to 9,000 mTorr, 2,500 to 10,000 mTorr, 2,500 to 11,000 mTorr, 2,500 to 12,000 mTorr, 2,500 to 13,000 mTorr, 2,500 to 14,000 mTorr, 2,500 to 15,000 mTorr, 2,500 to 20,000 mTorr, 2,500 to 25,000 mTorr, 2,500 to 30,000 mTorr, 2,500 to 35,000 mTorr, 2,500 to 40,000 mTorr, 2,500 to 45,000 mTorr, 2,500 to 50,000 mTorr, 2,500 to 55,000 mTorr, or 2,500 to 60,000 mTorr.

[0027] When outside the above range, damage to the protein may occur due to phenomena such as boiling, and there is a problem that a perfect structure cannot be obtained because sufficient drying does not proceed.

[0028] On the other hand, in the case of the vacuum drying, it is necessary to reach a constant rate in a state where it is stably controlled to the target pressure at normal atmospheric pressure which is a general atmospheric condition.

[0029] In yet another embodiment of the present invention, the method for producing the dried cake of botulinum toxin of the present invention may be produced by vacuum drying the above-described botulinum toxin at a temperature of 3 to 25°C.

[0030] In a specifically preferred embodiment, the vacuum drying of the present invention may be performed at a temperature of 3°C to 25°C, 5°C to 25°C, 7°C to 25°C, 9°C to 25°C, 11°C to 25°C, 12°C to 25°C, 3°C to 20°C, 5°C to 20°C, 7°C to 20°C, 9°C to 20°C, 11°C to 20°C, 12°C to 20°C, 3°C to 18°C, 3°C to 16°C, 3°C to 14°C, or 3°C to 12°C.

[0031] When outside the above range, it is possible to increase the structural instability of the botulinum toxin depending on the high or low temperature, or to cause physical damage and deformation, and there is a problem that a reduction in efficacy may occur thereby.

[0032] In yet another embodiment of the present invention, the method for producing the dried cake of botulinum toxin of the present invention may be performed until the moisture content of the dried cake of botulinum toxin of the present invention reaches within 3%.

[0033] Specifically, the method for producing the dried cake of botulinum toxin of the present invention may be produced by vacuum drying the above-described botulinum toxin for 0.5 hours or more, preferably between 0.5 hours and 4 hours.

[0034] In a specifically preferred embodiment, the vacuum drying of the present invention may be carried out for a period of 0.5 to 4 hours, 0.5 to 3 hours, 0.5 to 2 hours, 0.5 to 1 hour, 1 to 4 hours, 2 to 4 hours, or 3 to 4 hours.

[0035] The final product produced by the method for producing botulinum toxin dried cake of the present invention, i.e., botulinum toxin dried cake, may have a standard titer of 80% to 120%, or 85% to 115%.

[0036] In the manufacturing process of the botulinum toxin dried cake of the present invention, depending on the volume or surface area of ​​the solvent used for drying the product, boiling over may occur due to changes in the amount of dissolved oxygen or the concentration of the composition concentrated during the drying process, which can change the freezing point and lead to the formation of unintended ice nuclei. If the drying of the product proceeds under such conditions, it can induce non-uniformity of the product's properties and a decrease in product strength due to protein damage. Therefore, it is preferable that the drying process proceeds under conditions in which the volume or surface area of ​​the solvent used for drying the product is appropriately maintained, and this is within a range that can be easily adjusted by an ordinary technician familiar with the contents of this specification.

[0037] The present invention provides a method for producing botulinum toxin dried cake using the vacuum drying process described above, thereby eliminating the freeze-drying process of the prior art. In one embodiment of the present invention, the present invention uses a vacuum drying process, thereby shortening the process time and minimizing damage to the protein pharmaceutical during drying, and thus the present invention can have advantages in terms of more efficient and commercially viable production compared to the prior art. [Effects of the Invention]

[0038] The present invention relates to a method for vacuum drying botulinum toxin. When using the vacuum drying method of the present invention to produce a dried botulinum toxin cake, the drying time can be significantly reduced while maintaining the efficacy of the botulinum toxin. [Brief explanation of the drawing]

[0039] [Figure 1] The results of comparing the cake properties of vacuum-dried and freeze-dried products are shown. [Figure 2a-b] The results of our investigation into the potential for denaturation and loss of botulinum toxin under typical low-vacuum, reduced-pressure drying conditions are shown below. [Figure 3a-b] The results of confirming the difference in cake properties (Figure 3a) and the change in vial temperature (Figure 3b) under pressure conditions ranging from 1,000 to 70,000 mTorr in the vacuum drying method of the present invention are shown. [Figure 4] The results of confirming the change in temperature inside the vial under pressure conditions in the range of 50,000 to 70,000 mTorr in the vacuum drying method of the present invention are shown. [Modes for carrying out the invention]

[0040] The present invention will be explained in more detail below through the examples. These examples are solely for the purpose of illustrating the present invention in more detail, and it will be obvious to those with ordinary skill in the art that the scope of the present invention is not limited by these examples, as is the essence of the invention.

[0041] Unless otherwise indicated in the examples, “toxin” or “botulinum toxin” means botulinum toxin type A complex having a molecular weight of approximately 900 kDa. The methods disclosed herein are applicable to preparations of toxins, complexes, botulinum toxin serotypes, and botulinum neurotoxin components of other molecular weights, as well as those of approximately 150 kDa, approximately 300 kDa, and approximately 500 kDa.

[0042] Preparation example: Preparation of the final stock solution of botulinum toxin. The final stock solution of botulinum toxin was prepared by adding 0.9% sodium chloride (Merck, 1.37017.5000), 0.5% human serum albumin (Green Cross, 161B19508), and 1000 U / mL botulinum toxin type A stock solution (Inibio Co., Ltd., South Korea).

[0043] Comparative Example 1. Comparison of botulinum toxin potency by freeze-drying. 1-1. Drying Experiment The formulations dried under freeze-drying conditions and under vacuum drying conditions were compared. The freeze-drying and vacuum drying conditions are shown in Table 1 below.

[0044] [Table 1]

[0045] As can be seen in Figure 1, freeze-drying (right vial in Figure 1) formed a thicker white dried cake compared to vacuum drying (left vial in Figure 1). Such dried cakes are prone to cracking due to impacts applied to the vial, and there is a possibility that the broken dried cake will scatter.

[0046] 1-2. Titer Experiment The final stock solution of botulinum toxin prepared in the above preparation example was dried under freeze-drying conditions (see Table 2 below). The prepared solution was dissolved in 2.8 mL of physiological saline and administered intraperitoneally (0.1 mL / mice) to 10 ICR-mice (4 weeks old, body weight 18-22 g; Coretech Co., Ltd., South Korea). The number of dead and surviving animals was checked for 3 days. The titer was calculated using a statistical program (CombiStats 6.1, EDQM). The appropriate titer standard was set to an interval that included the error range of the animal study (see Table 3 below), and the results are shown in Table 4 below.

[0047] [Table 2]

[0048] [Table 3]

[0049] [Table 4]

[0050] As can be seen from Table 4, the potency of botulinum toxin preparations that have undergone a typical, conventional freeze-drying process is 56-65%, which is outside the range listed in Table 3, indicating that the effect of protein damage has been reduced.

[0051] These results indicate that the vacuum-dried form of botulinum toxin can maintain its activity more effectively than the freeze-dried form.

[0052] Reference example: Confirmation of denaturation and loss of botulinum toxin by vacuum drying. The final stock solution of botulinum toxin prepared in the above preparation example was filled into vials in 5 mL or 1 mL increments, and the physical changes and freezing point changes due to typical low-vacuum drying pressure changes were observed.

[0053] Specifically, vacuum drying was performed by varying the pressure from 1 bar to 0.0019 bar (approximately 1500 mTorr). The shelf temperature was maintained at 5°C. Distilled water (DW) and a 0.9% NaCl solution were used as control groups.

[0054] Figure 2a shows the phase change observed in response to pressure after filling vials with 5 mL of each solution. Under the conditions described above, boiling of the final stock solution of botulinum toxin occurred at pressures below 10,000 mTorr, but not in the control group.

[0055] Figure 2b shows the phase changes observed in response to pressure after filling vials with 1 mL of each solution. Under the conditions described above, boiling of the final stock solution and formation of ice crystals occurred at 1,500 mTorr, but not in the control group.

[0056] Such phenomena are understood to occur because the volume or surface area of ​​the solvent is not suitable for the solvent's properties and drying conditions. This can make it difficult to properly control the drying process, or cause sublimation instead of complete evaporation due to the formation of ice crystals during the process, making it difficult to achieve normal vacuum drying. Furthermore, if the drying of the product proceeds under such conditions where proper control is impossible, it can induce non-uniformity of product properties and a decrease in product strength due to protein damage, etc.

[0057] Example 1. Establishment of vacuum drying conditions through confirmation of botulinum toxin titer. The final stock solution of botulinum toxin prepared in the above preparation example was filled into vials in 0.1 mL increments, and the formulations were dried under reduced pressure under the conditions described in Table 5 below. The potency was calculated using the same method as in Comparative Examples 1-2. The appropriate potency standard was set to an interval that included the error range of the animal test (see Table 3 above), and the results are shown in Table 5 below.

[0058] [Table 5]

[0059] As can be seen from Table 5, the potency of botulinum toxin preparations dried at 1,500-30,000 mTorr and 3-25°C for 1 hour met the range described in Table 3. In contrast, it was found that when the drying process was performed outside the range of 1,500-30,000 mTorr (Comparative Examples 4 and 5), the potency of the botulinum toxin preparations did not meet the acceptable range.

[0060] Example 2. Confirmation of drying time under the vacuum drying conditions of the present invention #1 Drying can consist of two main steps: separation of water molecules linked by hydrogen bonds, and removal of water molecules that are interacting in various ways with other excipients and proteins that make up the final stock solution. When water molecules are bonded to each other, separation is possible with relatively less energy than when water molecules are interacting with other molecules. In the majority of cases, water molecules are bonded to each other, and this separation leads to primary evaporation. During this process, the liquid product undergoes a transformation into a gaseous state. Secondary evaporation occurs when the bonds between water molecules and other excipients are broken, and drying is considered complete when this process is almost finished. The moisture content must be 3% or less to meet the standards for dried cake injection.

[0061] Therefore, the rate of water evaporation and suitability can be confirmed through the phase change of the product.

[0062] 2-1. Confirmation of phase change (liquid → gas) time under different pressure conditions Based on the results of Example 1, 0.1 mL each of the empty vial and the final stock solution of botulinum toxin prepared in the above preparation example were filled into the vial, and the phase change time under different pressure conditions was confirmed.

[0063] Specifically, as shown in Table 6 below, a drying process was carried out for 1 hour under conditions of 1,000 to 70,000 mTorr and 25°C, and a temperature sensor was placed inside the vial to confirm the temperature change due to phase change. The rate at which phase change occurs is faster at lower pressures, and through this, it can be confirmed that evaporation is occurring.

[0064] [Table 6]

[0065] As can be seen from Table 6 and Figures 3a and 3b, a white, dry cake formed at the bottom of most vials, indicating a phase change due to evaporation, with the time required for this phase change being 11 to 43 minutes. However, this process did not appear within one hour at 50,000 mTorr, 60,000 mTorr, and 70,000 mTorr.

[0066] These results suggest that the time required for complete drying (evaporation) increases with increasing pressure, and that it is difficult to produce the botulinum toxin dried cake targeted by this invention at pressures exceeding a certain range (Comparative Example 7).

[0067] 2-2. Confirmation of the moisture content of the manufactured botulinum toxin dried cake. Based on the results of Examples 1 and 2-1, the moisture content of vials containing botulinum toxin dried cake, which was prepared by filling empty vials with 0.1 mL each of the final stock solution of botulinum toxin prepared in the above preparation example and drying under reduced pressure at 3,500 mTorr and 5°C, was measured in an environment with a humidity of 10% or less. The humidity content (%) was calculated using the following formula, and the appropriate humidity standards were shown in Table 7 below, with reference to the pharmacopoeia.

[0068]

number

[0069] [Table 7]

[0070] [Table 8]

[0071] As can be seen from Table 8, the humidity of the botulinum toxin preparations that underwent a drying process for 0.5 to 4 hours met the ranges described in Table 7.

[0072] Example 3. Confirmation of drying time under the vacuum drying conditions of the present invention #2 Based on the results of Example 2, 0.1 mL each of the empty vial and the final stock solution of botulinum toxin prepared in the above preparation example were filled into the vial, and the phase change time under different pressure conditions was confirmed.

[0073] Specifically, as shown in Table 9 below, a drying process was carried out for 3 hours under conditions of 50,000-70,000 mTorr and 25°C, and a temperature sensor was placed inside the vial to confirm the temperature change due to phase change. The rate at which phase change occurs is faster at lower pressures, and through this, it can be confirmed that evaporation is occurring.

[0074] [Table 9]

[0075] As can be seen from Table 9 and Figure 4, the time required for the phase change to occur under 50,000 and 60,000 mTorr conditions was 143 minutes and 183 minutes, respectively. However, the relevant process did not appear at 70,000 mTorr.

[0076] These results suggest that the time required for complete drying (evaporation) increases with increasing pressure, and that it is difficult to produce the botulinum toxin dried cake targeted by this invention at pressures exceeding a certain range (Comparative Example 8).

Claims

1. The process includes the step of drying the botulinum toxin under reduced pressure conditions of 1,500 mTorr to 60,000 mTorr. A method for producing a dried botulinum toxin cake, characterized in that the vacuum drying is carried out at a temperature of 3°C to 25°C for 0.5 hours to 4 hours.

2. The method for producing a dried botulinum toxin cake according to claim 1, characterized in that the concentration of the botulinum toxin is 250 U / mL to 5,000 U / mL.

3. A method for producing a dried botulinum toxin cake according to claim 1 or 2, characterized in that the produced dried botulinum toxin cake has a standard potency of 80% to 120% or 85% to 115%.

4. The method for producing a dried botulinum toxin cake according to claim 1 or 2, characterized in that the reduced-pressure drying is carried out until the humidity content of the dried botulinum toxin cake reaches 3% or less.

5. The method for producing a dried botulinum toxin cake according to claim 1 or 2, characterized in that the botulinum toxin-producing bacterial strain is Clostridium botulinum Type A.

6. The method for producing a dried botulinum toxin cake according to claim 1 or 2, characterized in that the pressure conditions for reduced-pressure drying are formed by reaching a constant speed while stably controlling the pressure from atmospheric pressure to a target pressure.

7. The aforementioned vacuum drying is performed at 1,500-60,000 mTorr, 1,500-55,000 mTorr, 1,500-50,000 mTorr, 1,500-45,000 mTorr, 1,500-40,000 mTorr, 1,500-35,000 mTorr, 1,500-30,000 mTorr, 1,500-25,000 mTorr, 1,500-20,000 mTorr, 1,500-15,000 mTorr, 1,500-14,000 mTorr, 1,500-13,000 mTorr, and 1,500- 12,000mTorr, 1,500-11,000mTorr, 1,500-10,000mTorr, 1,500-9,000mTorr, 1,500-8,000mTorr, 1,500-7,000mTorr, 1,500-6,000mTorr , 1,500-5,000mTorr, 1,500-4,000mTorr, 1,500-3,500mTorr, 1,500-3,000mTorr, 1,500-2,500mTorr, 1,500-2,000mTorr, 2,500-3,000m Torr, 2,500-3,500mTorr, 2,500-4,000mTorr, 2,500-4,500mTorr, 2,500-5,000mTorr, 2,500-6,000mTorr, 2,500-7,000mTorr, 2,500-8, 000mTorr, 2,500-9,000mTorr, 2,500-10,000mTorr, 2,500-11,000mTorr, 2,500-12,000mTorr, 2,500-13,000mTorr, 2,500-14,000mTorr A method for producing a dried botulinum toxin cake according to claim 1 or 2, characterized in that the process is carried out at a pressure of 2,500 to 15,000 mTor, 2,500 to 20,000 mTor, 2,500 to 25,000 mTor, 2,500 to 30,000 mTor, 2,500 to 35,000 mTor, 2,500 to 40,000 mTor, 2,500 to 45,000 mTor, 2,500 to 50,000 mTor, 2,500 to 55,000 mTor, or 2,500 to 60,000 mTor.

8. The method for producing a dried botulinum toxin cake according to claim 1 or 2, characterized in that the vacuum drying is carried out at a temperature of 5°C to 25°C, 7°C to 25°C, 9°C to 25°C, 11°C to 25°C, 12°C to 25°C, 3°C to 20°C, 5°C to 20°C, 7°C to 20°C, 9°C to 20°C, 11°C to 20°C, 12°C to 20°C, 3°C to 18°C, 3°C to 16°C, 3°C to 14°C, or 3°C to 12°C.

9. The method for producing a dried botulinum toxin cake according to claim 1 or 2, characterized in that the vacuum drying is carried out for a period of 0.5 to 4 hours, 0.5 to 3 hours, 0.5 to 2 hours, 0.5 to 1 hour, 1 to 4 hours, 2 to 4 hours, or 3 to 4 hours.

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