Stable defibrating enzyme preparation

Abstract

The invention provides a stable defibrase preparation. The defibrase preparation consists of defibrase, dextran 40, trehalose and a solvent for injection. The invention also provides a preparation method and a quality control method of the injection, the preparation is simple in prescription, the prepared freeze-dried preparation is better in stability during storage and convenient to use, and compared with the injection prepared from defibrase, the stability is better.

Description

Technical Field

[0001] The invention relates to the pharmaceutical field, in particular to a stable defibrase preparation. Background Art

[0002] Defibrase for injection is used for acute cerebral infarction, including cerebral thrombosis, cerebral embolism, transient ischemic attack (TIA), and prevention of recurrence of cerebral infarction. Myocardial infarction, unstable angina pectoris and prevention of recurrence of myocardial infarction. Limb vascular disease, including femoral artery embolism, thromboangiitis obliterans, Raynaud's disease, blood in a hyperviscosity state, hypercoagulable state, and pre-thrombotic state. Summary of the invention

[0003] The invention aims to provide a defibrase preparation with simple formula, scientific characteristics and good stability.

[0004] To achieve the above-mentioned purpose of the invention, the present invention provides a stable defibrase preparation, and the specific implementation scheme is as follows: The invention discloses a stable defibrase preparation, which consists of a defibrase solution, dextran 40, trehalose and an injection solvent.

[0005] The invention discloses a stable defibrase preparation, wherein the defibrase is a proteolytic enzyme extracted from the venom of Agkistrodon halys or Agkistrodon acutus in Changbai Mountain.

[0006] The stable defibrase preparation of the present invention, the injection solvent is water for injection.

[0007] The stable defibrase preparation of the present invention has a formula weight percentage of: Defibrase 5000 units or 10000 units Dextran 40 1-10 parts 1-10 parts of trehalose Add water for injection to 1000ml.

[0008] The stable defibrase preparation of the present invention has a formula weight percentage of: Defibrase 5000 units or 10000 units Dextran 40 5 parts 5-10 parts of trehalose Add water for injection to 1000ml.

[0009] The present invention discloses a stable defibrase preparation, wherein the preparation method is as follows: 1) Weigh an appropriate amount of water for injection into the preparation tank; 2) Add dextran 40 and trehalose to the batching tank, stir to dissolve completely, add defibrase and then add water for injection. The solution temperature should not be higher than 20℃; measure the potency and determine the filling amount; 3) Filter with 0.45μm and 0.25μm microporous membranes, and package after the clarity of the solution is qualified; 4) freeze-dry the solution after packaging in step 3), cool it to -40°C within 1 hour, pre-freeze it for 1-2 hours, heat it to -30°C within 1 hour, and then heat it to -20°C at a slope of 0.5°C~1°C, continue sublimation drying, and the final drying temperature is not higher than 20°C.

[0010] 5) Capping, rolling, packaging, full inspection and warehousing.

[0011] The stable defibrase preparation of the present invention has a quality control method as follows: 1) Properties, including observation of appearance and solubility; 2) Identification items, using two methods: coagulation reaction identification and chemical identification; 3) Inspection items, including acidity, potency, loss on drying, hemorrhagic toxicity, abnormal toxicity, filling volume difference, allergy test, bacterial endotoxin and sterility test; Analysis of the beneficial effects of the formulation of the present invention: the injection of defibrase is mainly small-volume injection and freeze-dried powder injection. Defibrase, as a protease, is unstable when prepared into freeze-dried preparations and injections. Polysaccharides such as dextran can increase the glass transition temperature of the product and prevent protein damage caused by product collapse. Glucose can partially remain amorphous during the freeze-drying process, and has a certain protective effect on certain protein drugs. It also has a very good scaffolding effect during the freeze-drying process and can obtain a good freeze-dried appearance. Trehalose has a higher glass transition temperature, lower hygroscopicity, and is less reducing, which can reduce the damage of the freeze-drying process to defibrase. The hydroxyl groups of trehalose form hydrogen bonds with the polar groups of proteins to form a hydration film around the protein, stabilize the protein structure, and prevent the protein from denaturing due to freeze-drying. Therefore, this patent selects dextran 40 and trehalose as protective agents for freeze-dried preparations. Implementation

[0012] The present invention is further illustrated by the following examples, but are not intended to limit the present invention. Example

[0013] Defibrase 5000 units Dextran 40 5g Trehalose 1g Add water for injection to 1000ml Produced 1000 bottles Its preparation process is: 1. Weigh an appropriate amount of water for injection and place it in the preparation tank, keeping the solution temperature no higher than 30°C; 2. Add 5g of dextran 40 and 1g of trehalose to the batching tank, stir to dissolve completely, lower the temperature to no more than 20°C, add defibrase and stir to dissolve, add water for injection to 1000ml, measure the potency, and determine the filling amount; 3. Filter with 0.45um and 0.25um microporous membranes, and pack the solution into 1ml bottles after the clarity is qualified. 4. Freeze drying: Quickly freeze and cool to -40℃ and maintain for 2 hours. Heat to -30℃ within 0.5h, then heat to -20℃ at a rate of 1℃ per hour, taking 11h, with a vacuum degree of 15Pa. Heat to -10℃ within 5h, then heat to 10℃ within 6h, with a vacuum degree of 15Pa. Finally, dry at 10℃ for 5 hours, with a vacuum degree of 30Pa.

[0014] 5. Capping, rolling, packaging, full inspection and warehousing. Example

[0015] Defibrase 5000 units Dextran 40 5g Trehalose 5g Add water for injection to 1000ml Produced 1000 bottles Its preparation process is: 1. Weigh an appropriate amount of water for injection and place it in the preparation tank, keeping the solution temperature no higher than 30°C; 2. Add 5g of dextran 40 and 5g of trehalose to the batching tank, stir to dissolve completely, lower the temperature to no more than 20°C, add defibrase and stir to dissolve, add water for injection to 1000ml, measure the potency and determine the filling amount; 3. Filter with 0.45um and 0.25um microporous membranes, and pack the solution into 1ml bottles after the clarity is qualified. 4. Freeze drying: Quickly freeze and cool to -40℃ and maintain for 2 hours. Heat to -30℃ within 0.5h, then heat to -20℃ at a rate of 1℃ per hour, taking 11h, with a vacuum degree of 15Pa. Heat to -10℃ within 5h, then heat to 10℃ within 6h, with a vacuum degree of 15Pa. Finally, dry at 10℃ for 5 hours, with a vacuum degree of 30Pa.

[0016] 5. Capping, rolling, packaging, full inspection and warehousing. Example

[0017] Defibrase 5000 units Dextran 40 5g Trehalose 10g Add water for injection to 1000ml Produced 1000 bottles Its preparation process is: 1. Weigh an appropriate amount of water for injection and place it in the preparation tank, keeping the solution temperature no higher than 30°C; 2. Add 5g of dextran 40 and 10g of trehalose to the batching tank, stir to dissolve completely, lower the temperature to no more than 20°C, add defibrase and stir to dissolve, add water for injection to 1000ml, measure the potency and determine the filling amount; 3. Filter with 0.45um and 0.25um microporous membranes, and pack the solution into 1ml bottles after the clarity is qualified. 4. Freeze drying: Quickly freeze and cool to -40℃ and maintain for 2 hours. Heat to -30℃ within 0.5h, then heat to -20℃ at a rate of 1℃ per hour. It takes 11h and the vacuum degree is 15Pa. Heat to -10℃ within 5h, then heat to 10℃ for 6h, and the vacuum degree is 15Pa. Finally, dry at 10℃ for 5 hours, and the vacuum degree is 30Pa.

[0018] 5. Capping, rolling, packaging, full inspection and warehousing.

[0019] Comparative Example 1: The defibrase preparation was prepared using the formulation and preparation method of Example 1 in the Chinese patent application number CN201810115670 as Comparative Example 1.

[0020] Comparative Example 2: As Comparative Example 2, a defibrase preparation was prepared using the formulation and preparation method of Example 1 in Chinese Patent Application No. CN200510044516.

[0021] The stability of the defibrase for injection of Examples 1-3 of the present invention and Comparative Examples 1-2 was studied, and the test results of the influencing factors are as follows: Table 1 Quality comparison of various embodiments

[0022] Table 2 Comparison of influencing factors of various embodiments (high temperature)

[0023] The above experimental results prove that Examples 1-3 are significantly better than the comparative example, and the stability of Examples 1-3 is better than that of the comparative example 1.

[0024] Although the present invention has been described in detail above by means of general description, specific implementation methods and experiments, it is obvious to those skilled in the art that some modifications or improvements can be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention are within the scope of protection claimed by the present invention.

Claims

1. A stable defibrase preparation, It is characterized in that The defibrase preparation consists of defibrase, trehalose and a solvent for injection.

2. A stable defibrase preparation according to claim 1, It is characterized in that The solvent for injection is water for injection.

3. A stable defibrase preparation according to claim 1, It is characterized in that The weight parts of the formulation are as follows: Defibrase 5000 units or 10000 units Dextran 40 1-10 parts 1-10 parts of trehalose Add water for injection to 1000ml.

4. A stable defibrase preparation according to claim 1, It is characterized in that The weight parts of the formulation are as follows: Defibrase 5000 units or 10000 units Dextran 40 5 parts 5-10 parts of trehalose Add water for injection to 1000ml.

5. A stable defibrase preparation according to any one of claims 1 to 4, It is characterized in that The defibrase is a proteolytic enzyme extracted from the venom of Agkistrodon agkistrodon.

6. A stable defibrase preparation according to any one of claims 1 to 5, It is characterized in that The preparation method of the preparation is: 1) Weigh an appropriate amount of water for injection into the preparation tank; 2) Add dextran 40 and trehalose to the batching tank, stir to dissolve completely, add defibrase and then add water for injection. The solution temperature should not be higher than 20℃; measure the potency and determine the filling amount; 3) Filter with 0.45μm and 0.25μm microporous membranes, and package after the clarity of the solution is qualified; 4) freeze-drying the solution after packaging in step 3), cooling to -40°C within 1 hour, pre-freezing for 1-2 hours, heating to -30°C within 1 hour, heating to -20°C at a slope of 0.5°C~1°C, and continuing sublimation drying, with the final drying temperature not higher than 20°C; 5) Capping, rolling, packaging, full inspection and warehousing.

7. A stable defibrase preparation according to claim 6, It is characterized in that The quality control method of this preparation is: 1) Properties, including observation of appearance and solubility; 2) Identification items, using two methods: coagulation reaction identification and chemical identification; 3) Inspection items, including acidity, potency, loss on drying, hemorrhagic toxicity, abnormal toxicity, filling volume difference, allergy test, bacterial endotoxin and sterility test.

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