NAD + freeze-dried powder injection preparation and preparation process thereof

Abstract

The invention belongs to the field of pharmaceutical preparations, and particularly relates to an NAD + freeze-dried powder injection preparation and a preparation process thereof. The freeze-dried powder injection preparation comprises NAD < + >, a freeze-drying protective agent, an auxiliary protective agent, a stabilizer and a buffer system. The NAD + freeze-dried powder injection preparation prepared by adding the freeze-drying protective agent, the auxiliary protective agent and the stabilizer, screening the types of the freeze-drying protective agent, the auxiliary protective agent and the stabilizer, optimizing the prescription composition of the freeze-dried powder injection preparation and simplifying the freeze-drying process is stable in quality and can be stored for a long time, meanwhile, the freeze-drying process is simple in temperature control and easy to operate, and industrial production is easy.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations and relates to a NAD + Freeze-dried powder injection preparation and its preparation process. Background Art

[0002] NAD + ), chemically known as nicotinamide adenine dinucleotide or nicotinamide diphosphate, NAD + It is an essential coenzyme in living organisms. It plays a role in transferring hydrogen in the biological oxidation process. It can activate various enzyme systems, promote the synthesis and metabolism of nucleic acids, proteins, and polysaccharides, increase material transport and regulation, and improve metabolic function. + It is clinically used as an auxiliary treatment for coronary heart disease to improve symptoms such as chest tightness and angina pectoris.

[0003] However, NAD + It is a small molecule polypeptide with unstable structure, easy to absorb moisture, easy to decompose when heated, and easy to deteriorate in alkaline solution. The commonly used dosage form is coenzyme I for injection, which is white or off-white freeze-dried block or powder, and is injected intramuscularly and dissolved with sodium chloride injection before use.

[0004] Patent CN105534927B provides a stable oxidized coenzyme I preparation and its preparation method, using lactose as a freeze-drying protective agent and a suitable freeze-drying process to prepare a stable oxidized coenzyme I preparation, but the temperature control operation during the freeze-drying process is relatively complicated. Therefore, it is necessary to develop a NAD lyophilization process that is simple and easy to operate, has stable quality, and has good storage stability. + Freeze-dried powder injection preparation. Summary of the invention

[0005] The main purpose of the present invention is to provide a NAD + Freeze-dried powder injection preparation and preparation process thereof, prepared NAD + The freeze-dried powder injection preparation has stable quality and good storage stability. The freeze-drying process temperature control is simple and easy to operate, and it is easy to industrialize production.

[0006] The present invention provides a NAD + A freeze-dried powder injection preparation, wherein the freeze-dried powder injection preparation comprises, by weight: NAD + 1 part, lyophilizing protective agent 5-20 parts, auxiliary protective agent 1-10 parts, stabilizer 0.5-8 parts, citric acid-sodium citrate amount.

[0007] Preferably, the lyophilized powder injection preparation comprises, by weight: NAD + 1 part, lyophilizing protective agent 8-15 parts, auxiliary protective agent 2-6 parts, stabilizer 1-5 parts, citric acid-sodium citrate amount.

[0008] Preferably, the lyoprotectant is selected from one or more of sucrose, trehalose, lactose, glucose, maltose, mannitol, and sorbitol. More preferably, the lyoprotectant is selected from trehalose and sucrose. Most preferably, the lyoprotectant is selected from trehalose.

[0009] Preferably, the auxiliary protective agent is selected from one or more of histidine, lysine, arginine, glycine, glutamic acid, and aspartic acid. More preferably, the auxiliary protective agent is selected from arginine and glycine. Most preferably, the auxiliary protective agent is selected from arginine.

[0010] Preferably, the stabilizer is selected from one or more of polyvinyl pyrrolidone, polyethylene glycol, and dextran. More preferably, the stabilizer is selected from polyvinyl pyrrolidone, and the polyvinyl pyrrolidone model is selected from K15, K25, K30, K60, and K90.

[0011] The present invention also provides a NAD + The preparation method of the freeze-dried powder injection preparation comprises the following steps:

[0012] The prescribed amount of lyophilized protective agent, auxiliary protective agent, stabilizer and NAD + Add an appropriate amount of citric acid-sodium citrate buffer, stir to dissolve, adjust the pH with citric acid or sodium hydroxide, make up the volume with citric acid-sodium citrate buffer, filter, fill, and freeze-dry; the freeze-drying comprises the following steps: cool the solution before freeze-drying to -50°C to -40°C, maintain for 1 to 3 hours; raise the temperature to -20°C to -10°C at 0.2-0.3 mbar, maintain for 15 to 20 hours; raise the temperature of the sample to 20 to 30°C, maintain for 2 to 6 hours, and finally obtain NAD + Freeze-dried powder injection preparation.

[0013] Preferably, the concentration of the citric acid-sodium citrate buffer is 10-50 mM.

[0014] Preferably, the pH is adjusted to 5.0-6.0 with citric acid or sodium hydroxide.

[0015] Compared with the prior art, the present invention has the following beneficial effects: the present invention optimizes the formulation composition of the freeze-dried powder injection preparation by adding freeze-drying protective agents, auxiliary protective agents, and stabilizers, and screens their types, simplifies the freeze-drying process, and prepares NAD + Freeze-dried powder injection preparations have stable quality and can be stored for a long time. At the same time, the temperature control of the freeze-drying process is simple and easy to operate, and it is easy to industrialize production. DETAILED DESCRIPTION

[0016] In view of the defects of the prior art, the inventor has proposed the technical solution of the present invention after long-term research and extensive practice. The present invention is further illustrated by embodiments below, but these embodiments do not constitute a limitation of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0017] Example 1 Screening of Lyoprotectant Types

[0018]

[0019]

[0020] Preparation method: freeze-dried protective agent, auxiliary protective agent, stabilizer and NAD + Add to 80% citric acid-sodium citrate buffer, stir to dissolve, adjust pH to 5.5 with citric acid solution or sodium hydroxide solution, and make up to volume with citric acid-sodium citrate buffer, where NAD + The concentration is 2.5 mg / ml; filtering, filling (specification 5 mg: 2 ml), and freeze drying; the freeze drying includes the following steps: cooling the solution before freeze drying to -45°C and maintaining for 2 hours; heating to -15°C at 0.2 mbar and maintaining for 18 hours; raising the temperature of the sample to 25°C for 4 hours, extreme vacuum, and finally obtaining NAD + Freeze-dried powder injection preparation.

[0021] The freeze-dried samples of F1-F5 were placed in a 40°C thermostat for 10 days, and then sampled for testing. The clarity, pH, and content were measured after being reconstituted with 2 ml of water for injection. The results are as follows:

[0022]

[0023] From the results in the table above, it can be seen that compared with other prescriptions, when F5 does not add lyophilization protective agent, the prepared lyophilized powder injection preparation will collapse, and there will be fine particles after reconstitution, the content will be reduced, and the stability will be reduced. In addition, compared with lactose and mannitol, the lyophilized powder injection preparation prepared by using trehalose and sucrose as lyophilization protective agents has a more stable quality when placed at 40°C for 10 days, which is better than the lyophilized products in the prior art, and trehalose has the best stability as a lyophilization protective agent.

[0024] Example 2 Screening of auxiliary protective agent types

[0025]

[0026] The preparation method is the same as that in Example 1.

[0027] The freeze-dried samples of F6-F9 were placed in a 40°C thermostat for 10 days, and samples were taken for testing. The testing method was the same as in Example 1, and the results were as follows:

[0028]

[0029] From the results in the above table, it can be seen that compared with other prescriptions, F9 does not add auxiliary protective agents, and the stability of the freeze-dried powder injection prepared by it is reduced after being placed under the influence of 40°C for 10 days. In addition, compared with histidine and aspartic acid, the freeze-dried powder injection prepared by using arginine and glycine as auxiliary protective agents is more stable in quality after being placed under the influence of 40°C for 10 days, which is better than the freeze-dried products in the prior art, and arginine has the best stability as an auxiliary protective agent.

[0030] Example 3 Stabilizer Type Screening

[0031]

[0032] The preparation method is the same as that in Example 1.

[0033] The freeze-dried samples of F10-F12 were placed in a 40°C thermostat for 10 days, and samples were taken for testing. The testing method was the same as in Example 1, and the results were as follows:

[0034]

[0035] From the results in the above table, it can be seen that compared with other prescriptions, the stability of the freeze-dried powder injection prepared by F12 without adding stabilizer and F11 adding dextran decreased after being placed under the influence of 40°C for 10 days. In addition, compared with PEG4000, the freeze-dried powder injection prepared by using polyvinyl pyrrolidone as a stabilizer was more stable after being placed under the influence of 40°C for 10 days. Therefore, polyvinyl pyrrolidone was used as a stabilizer.

[0036] Example 4 Stability Study

[0037]

[0038] The preparation method is the same as that in Example 1.

[0039] The freeze-dried samples of F1-F2, F8, F13-F18 were placed in a 40°C constant temperature box for 10 days, and placed in a 25°C / 60% RH constant temperature and humidity box for 24 months. The samples were taken for testing. The testing method was the same as that in Example 1. The results are as follows:

[0040]

[0041]

[0042] From the results in the table above, it can be seen that the NAD prepared by using trehalose and sucrose as freeze-drying protective agents, arginine and glycine as auxiliary protective agents, and polyvinyl pyrrolidone as a stabilizer + The freeze-dried powder injection preparation has good stability when placed at 40°C for 10 days, and has stable quality when stored at 25°C for two years.

[0043] It should be understood that the technical solution of the present invention is not limited to the above-mentioned specific implementation cases. Any technical deformation made according to the technical solution of the present invention without departing from the scope of protection of the purpose of the present invention and the claims shall fall within the protection scope of the present invention.

Claims

1. A type of NAD + The freeze-dried powder injection preparation is characterized in that: The freeze-dried powder injection preparation comprises, by weight: NAD + 1 part, lyophilization protective agent 5-20 parts, auxiliary protective agent 1-10 parts, stabilizer 0.5-8 parts, citric acid-sodium citrate appropriate amount; the lyophilization protective agent is selected from trehalose and sucrose, the auxiliary protective agent is selected from arginine and glycine, and the stabilizer is selected from polyvinyl pyrrolidone.

2. NAD according to claim 1 + The freeze-dried powder injection preparation is characterized in that: The freeze-dried powder injection preparation comprises, by weight: NAD + 1 part, lyophilizing protective agent 8-15 parts, auxiliary protective agent 2-6 parts, stabilizer 1-5 parts, citric acid-sodium citrate amount.

3. NAD according to claim 2 + The freeze-dried powder injection preparation is characterized in that: The freeze-dried powder injection preparation comprises, by weight: NAD + 1 part, lyophilizing protective agent 11.5 parts, auxiliary protective agent 4 parts, stabilizer 2.5 parts, citric acid-sodium citrate amount.

4. The NAD according to any one of claims 1 to 3 + The freeze-dried powder injection preparation is characterized in that: The lyoprotectant is selected from trehalose.

5. The NAD according to any one of claims 1 to 3 + The freeze-dried powder injection preparation is characterized in that: The auxiliary protective agent is selected from arginine.

6. The NAD according to any one of claims 1 to 3 + The freeze-dried powder injection preparation is characterized in that: The polyvinyl pyrrolidone model is selected from K15, K25, K30, K60, and K90.

7. NAD according to claim 5 + The freeze-dried powder injection preparation is characterized in that: The polyvinyl pyrrolidone model is selected from K25.

8. NAD according to any one of claims 1 to 7 + The preparation method of freeze-dried powder injection comprises the following steps: adding a prescribed amount of freeze-dried protective agent, auxiliary protective agent, stabilizer and NAD + Add an appropriate amount of citric acid-sodium citrate buffer, stir to dissolve, adjust the pH with citric acid or sodium hydroxide, make up the volume with citric acid-sodium citrate buffer, filter, fill, and freeze-dry; the freeze-drying comprises the following steps: cool the solution before freeze-drying to -50°C to -40°C, maintain for 1 to 3 hours; raise the temperature to -20°C to -10°C at 0.2-0.3 mbar, maintain for 15 to 20 hours; raise the temperature of the sample to 20 to 30°C, maintain for 2 to 6 hours, and finally obtain NAD + Freeze-dried powder injection preparation.

9. The preparation method according to claim 8, characterized in that: The concentration of the citric acid-sodium citrate buffer is 10-50 mM.

10. The preparation method according to claim 8, characterized in that: Adjust pH to 5.0-6.0 with citric acid or sodium hydroxide.

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