Search

JP-7855814-B2 - Drying process of botulinum toxin preparations

JP7855814B2JP 7855814 B2JP7855814 B2JP 7855814B2JP-7855814-B2

Inventors

  • キム チョンセ
  • イム ヒョンア
  • キム ヨンジェ
  • アン ヨンドク

Assignees

  • イニバイオ カンパニー リミテッド
  • テウン カンパニー リミテッド

Dates

Publication Date
20260511
Application Date
20220726
Priority Date
20211111

Claims (9)

  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 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.

Description

This invention relates to a drying process for toxin preparations, and more specifically, to a method for producing botulinum toxin dried cake by a vacuum drying method. Botulinum toxin (BTX) is a neurotoxin produced by the anaerobic bacterium Clostridium botulinum (C. botulinum). There are seven types (A-G), and currently, two types, Botulinum A (BTX-A) and B (BTX-B), are purified and used medically. Botulinum toxin works by inhibiting the release of the neurotransmitter acetylcholine, thereby blocking muscle contraction signaling and causing muscle relaxation. Specifically, it inhibits the release of acetylcholine, a neurotransmitter secreted at the presynaptic terminal at the neuromuscular junction, inducing nerve paralysis. While fillers are medical devices that fill areas with insufficient skin volume with a specific substance, botulinum toxin preparations are pharmaceuticals containing components that reduce muscle use by preventing the release of neurotransmitters that cause muscle contraction. Botulinum toxin is primarily used to reduce or eliminate frown lines and wrinkles around the eyes, but its indications are gradually expanding as it is also used to treat upper limb rigidity from stroke, eyelid spasms, and equinus deformities. 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 through a freeze-drying process. Examples include Meditoxin®, Xeomin®, and Dysport®. On the other hand, while numerous conventional freeze-drying methods for 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, thus taking a considerably long time (approximately 18 to 48 hours). In particular, the freezing process, which is essential to freeze-drying, can lead to the formation of ice nuclei or, due to the partial imbalance of excipient concentrations that occurs during freezing, it can cause a decrease in activity through damage to the protein structure. While this type of 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. 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 both primary and secondary drying. The results of comparing the cake properties of vacuum-dried and freeze-dried products are shown.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.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.The results of observing the change in temperature inside the vial under pressure conditions ranging from 50,000 to 70,000 mTorr in the vacuum drying method of the present invention are shown. 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 more concretely, 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. Unless otherwise specified in the examples, “toxin” or “botulinum toxin” refers to 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. Preparation Example: Preparation of 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). Comparative Example 1. Comparison of botulinum toxin potency by freeze-drying 1-1. Drying experiment Preparations dried under freeze-drying conditions and reduced-pressure drying conditions were compared. The freeze-drying conditions and reduced-pressure drying conditions are shown in Table 1 below. 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 a dried cake is pro