A type of NAD + Lyophilized powder injection formulations and their preparation process

Abstract

This invention belongs to the field of pharmaceutical preparations, specifically relating to an NAD+ formulation. + Lyophilized powder for injection and its preparation process. This lyophilized powder for injection includes NAD+. + This invention utilizes lyophilization protectants, auxiliary protectants, stabilizers, and buffer systems. By adding lyophilization protectants, auxiliary protectants, and stabilizers, and screening their types, the formulation composition of lyophilized powder injections is optimized, and the lyophilization process is simplified to prepare NAD2. + Freeze-dried powder injections have stable quality, can be stored for a long time, and the freeze-drying process is simple and easy to operate with temperature control, making it easy to industrialize.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical formulation technology and relates to an NAD3 inhibitor. + Lyophilized powder injection formulations and their preparation process. Background Technology

[0002] Coenzyme I (NAD) + Its chemical name is nicotinamide adenine dinucleotide or nicotinamide diphosphate, NAD+ + It is an essential coenzyme in living organisms, playing a role in hydrogen transfer during biological oxidation. It can activate various enzyme systems, promote the synthesis and metabolism of nucleic acids, proteins, and polysaccharides, increase substance transport and regulation, and improve metabolic function. + Clinically, it is used as an adjunct treatment for coronary heart disease to improve symptoms such as chest tightness and angina.

[0003] However, NAD + It is a small molecule polypeptide with an unstable structure, is highly hygroscopic, easily decomposes upon heating, and is extremely prone to deterioration in alkaline solutions. The commonly used dosage form is Coenzyme I for injection, which is a white or off-white lyophilized block or powder, administered via intramuscular injection, and dissolved in sodium chloride injection solution just before use.

[0004] Patent CN105534927B provides a stable oxidized coenzyme I preparation and its preparation method. It uses lactose as a lyophilization protectant and employs a suitable lyophilization process to prepare a stable oxidized coenzyme I preparation. However, the temperature control during the lyophilization process is relatively complex. Therefore, there is a need to develop a NAD+ preparation with a simple and easy-to-operate lyophilization process, stable quality, and good storage stability. + Lyophilized powder for injection. Summary of the Invention

[0005] The main objective of this invention is to provide a NAD + Lyophilized powder injection formulations and their preparation process, and the preparation of NAD3 + Freeze-dried powder injections have stable quality and good storage stability. The freeze-drying process is simple and easy to operate with temperature control, making it easy to industrialize.

[0006] This invention provides an NAD + Lyophilized powder for injection, wherein the lyophilized powder for injection comprises, by weight: NAD + 1 part, 5-20 parts freeze-drying protectant, 1-10 parts auxiliary protectant, 0.5-8 parts stabilizer, and appropriate amount of citric acid-sodium citrate.

[0007] Preferably, the lyophilized powder injection formulation comprises, by weight: NAD + 1 part, 8-15 parts freeze-drying protectant, 2-6 parts auxiliary protectant, 1-5 parts stabilizer, and appropriate amount of citric acid-sodium citrate.

[0008] Preferably, the freeze-drying protectant is selected from one or more of sucrose, trehalose, lactose, glucose, maltose, mannitol, and sorbitol. More preferably, the freeze-drying protectant is selected from trehalose and sucrose. Most preferably, the freeze-drying protectant 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 polyvinylpyrrolidone, polyethylene glycol, and dextran. More preferably, the stabilizer is selected from polyvinylpyrrolidone, and the polyvinylpyrrolidone type is selected from K15, K25, K30, K60, and K90.

[0011] This invention also provides an NAD + The preparation method of lyophilized powder injection includes the following steps:

[0012] The prescription amounts of lyophilization protectant, auxiliary protectant, stabilizer, and NAD were added. + Add an appropriate amount of citrate-sodium citrate buffer solution, stir to dissolve, adjust the pH with citrate or sodium hydroxide, bring the volume to a final volume with the citrate-sodium citrate buffer solution, filter, fill, and freeze-dry. The freeze-drying process includes the following steps: cooling the solution before freeze-drying to -50℃ to -40℃ and maintaining it for 1 to 3 hours; raising the temperature to -20℃ to -10℃ at 0.2-0.3 mbar and maintaining it for 15 to 20 hours; raising the temperature of the sample to 20 to 30℃ and maintaining it for 2 to 6 hours under ultimate vacuum, finally obtaining NAD. + Lyophilized powder for injection.

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

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

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: The present invention optimizes the formulation composition of lyophilized powder injection by adding lyophilization protectants, auxiliary protectants, and stabilizers, and by screening their types, thus simplifying the lyophilization process and obtaining NAD2. + Freeze-dried powder injections have stable quality, can be stored for a long time, and the freeze-drying process is simple and easy to operate with temperature control, making it easy to industrialize. Detailed Implementation

[0016] In view of the deficiencies of the prior art, the inventors, through long-term research and extensive practice, have proposed the technical solution of this invention. The invention is further illustrated below by means of embodiments, but these embodiments do not constitute a limitation of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without inventive effort are within the scope of protection of this invention.

[0017] Example 1: Screening of Lyophilization Protectants

[0018]

[0019]

[0020] Preparation method: The prescribed amounts of lyophilization protectant, auxiliary protectant, stabilizer, and NAD are added... + Add to 80% citrate-sodium citrate buffer, stir to dissolve, adjust pH to 5.5 with citrate solution or sodium hydroxide solution, and bring to volume with citrate-sodium citrate buffer. The NAD content should be... + The concentration was 2.5 mg / ml; the solution was filtered, filled (5 mg: 2 ml), and freeze-dried. The freeze-drying process included the following steps: cooling the solution to -45°C and maintaining it for 2 hours; heating it to -15°C at 0.2 mbar and maintaining it for 18 hours; raising the temperature of the sample to 25°C and maintaining it for 4 hours under ultimate vacuum, finally obtaining NAD. + Lyophilized powder for injection.

[0021] The lyophilized samples F1-F5 were placed in a 40℃ incubator for 10 days, and samples were taken for testing. Clarity, pH, and content were determined after reconstitution with 2 ml of water for injection. The results are as follows:

[0022]

[0023] The results in the table above show that, compared to other formulations, the lyophilized powder injection prepared with F5 without the addition of a lyophilization protectant collapses, exhibits fine particles after reconstitution, and shows reduced content and stability. Furthermore, compared to lactose and mannitol, the lyophilized powder injection prepared using trehalose and sucrose as lyophilization protectants shows greater quality stability after 10 days at 40°C, surpassing existing lyophilized products, with trehalose demonstrating the best stability as a lyophilization protectant.

[0024] Example 2: Screening of Auxiliary Protective Agents

[0025]

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

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

[0028]

[0029] The results in the table above show that, compared to other formulations, F9, without the addition of auxiliary protective agents, exhibits reduced stability after 10 days of storage at 40°C. Furthermore, compared to histidine and aspartic acid, lyophilized powder injections prepared using arginine and glycine as auxiliary protective agents demonstrate greater quality stability after 10 days of storage at 40°C, surpassing existing lyophilized products. Arginine shows the best stability among auxiliary protective agents.

[0030] Example 3: Screening of Stabilizer Types

[0031]

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

[0033] The freeze-dried samples F10-F12 were placed in a 40℃ constant temperature incubator for 10 days, and samples were taken for testing. The testing method was the same as in Example 1, and the results are as follows:

[0034]

[0035] The results in the table above show that, compared to other formulations, the lyophilized powder injections prepared by F12 (without a stabilizer) and F11 (with dextran) exhibited reduced stability after 10 days of storage at 40°C. Furthermore, compared to PEG4000, the lyophilized powder injections prepared using polyvinylpyrrolidone (PVP) as a stabilizer showed greater stability after 10 days of storage at 40°C. Therefore, PPVP is recommended as the stabilizer.

[0036] Example 4 Stability Study

[0037]

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

[0039] Freeze-dried samples F1-F2, F8, and F13-F18 were placed in a 40℃ constant temperature incubator for 10 days and then in a 25℃ / 60%RH constant temperature and humidity incubator for 24 months. Samples were taken for testing, and the determination method was the same as in Example 1. The results are as follows:

[0040]

[0041]

[0042] The results in the table above show that, using trehalose and sucrose as freeze-drying protectants, arginine and glycine as auxiliary protectants, and polyvinylpyrrolidone as a stabilizer, the prepared NAD3... + The lyophilized powder injection preparation showed good stability after being placed at 40℃ for 10 days, and maintained stable quality after being stored at 25℃ for two years.

[0043] It should be understood that the technical solutions of the present invention are not limited to the specific embodiments described above. Any technical modifications made to the technical solutions of the present invention without departing from the spirit and scope of the claims are within the scope of protection of the present invention.

Claims

1. A type of NAD + Lyophilized powder for injection, characterized in that, The lyophilized powder injection formulation comprises, by weight, NAD: + 1 part, 5-20 parts of freeze-drying protectant, 1-10 parts of auxiliary protectant, 0.5-8 parts of stabilizer, and an appropriate amount of citric acid-sodium citrate; the freeze-drying protectant is selected from trehalose and sucrose, the auxiliary protectant is selected from arginine and glycine, and the stabilizer is selected from polyvinylpyrrolidone.

2. The NAD according to claim 1 + Lyophilized powder for injection, characterized in that, The lyophilized powder injection formulation comprises, by weight, NAD: + 1 part, 8-15 parts freeze-drying protectant, 2-6 parts auxiliary protectant, 1-5 parts stabilizer, and appropriate amount of citric acid-sodium citrate.

3. The NAD according to claim 2 + Lyophilized powder for injection, characterized in that, The lyophilized powder injection formulation comprises, by weight, NAD: + 1 part, 11.5 parts freeze-drying protectant, 4 parts auxiliary protectant, 2.5 parts stabilizer, and appropriate amount of citric acid-sodium citrate.

4. The NAD according to any one of claims 1-3 + Lyophilized powder for injection, characterized in that, The freeze-drying protectant is selected from trehalose.

5. The NAD according to any one of claims 1-3 + Lyophilized powder for injection, characterized in that, The auxiliary protective agent is selected from arginine.

6. The NAD according to any one of claims 1-3 + Lyophilized powder for injection, characterized in that, The polyvinylpyrrolidone model is selected from K15, K25, K30, K60, and K90.

7. The NAD according to claim 5 + Lyophilized powder for injection, characterized in that, The polyvinylpyrrolidone is selected from model K25.

8. The NAD according to any one of claims 1-7 + The preparation method of lyophilized powder for injection includes the following steps: mixing the prescribed amounts of lyophilization protectant, auxiliary protectant, stabilizer, and NAD+. + Add an appropriate amount of citrate-sodium citrate buffer solution, stir to dissolve, adjust the pH with citrate or sodium hydroxide, bring the volume to a final volume with the citrate-sodium citrate buffer solution, filter, fill, and freeze-dry. The freeze-drying process includes the following steps: cooling the solution before freeze-drying to -50℃ to -40℃ and maintaining it for 1 to 3 hours; raising the temperature to -20℃ to -10℃ at 0.2-0.3 mbar and maintaining it for 15 to 20 hours; raising the temperature of the sample to 20 to 30℃ and maintaining it for 2 to 6 hours under ultimate vacuum, finally obtaining NAD. + Lyophilized powder for injection.

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

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

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