A kind of vitamin C freeze-dried powder injection for injection and preparation method thereof

Abstract

The present invention provides a vitamin C freeze-dried powder injection for injection and a preparation method thereof, and belongs to the technical field of pharmaceutical preparations. The vitamin C freeze-dried powder injection for injection of the present invention is prepared into 1000 bottles by including the following components: 900-1100g of vitamin C, 650-800g of excipient, 250-500g of L-cysteine, 120-250g of tert-butyl alcohol, and 300-500g of β-cyclodextrin. The present invention utilizes components such as L-cysteine, tert-butyl alcohol, and β-cyclodextrin to change the eutectic point temperature, glass transition temperature, and collapse temperature of vitamin C, significantly shorten the freeze-drying time and the re-dissolution time, and effectively prevent the degradation, discoloration, and content reduction of vitamin C.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and in particular to a vitamin C freeze-dried powder injection for injection and a preparation method thereof. Background Art

[0002] Vitamin C is a water-soluble vitamin that is sensitive to temperature and easily oxidized. Currently, the products on the market are mainly water injections (injections). Due to its poor stability, it is easy to oxidize and discolor during storage. A large amount of antioxidants and preservatives need to be added to the water injection dosage form. The State Food and Drug Administration is increasingly cautious about the use of antioxidants in injections, requiring that they be added as little as possible. Even if antioxidants are added, vitamin C water injections are still affected by the external environment, with unstable pH values, easy degradation of related substances, rapid decrease in content, and easy discoloration. Strict storage conditions are required, including protection from light, sealing, and storage at low temperatures. Powder injections are more stable.

[0003] Freeze-dried powder injection has the following advantages: 1. The drug is more stable: Freeze-dried powder injection is made by dissolving the main drug, filtering it through sterilization, and then packaging it through freeze-sublimation drying under low-temperature sterile vacuum conditions. During the preparation of the liquid medicine, nitrogen is filled, and after freeze drying, nitrogen is filled and sealed. The effect of oxygen on the product can be effectively controlled during the production process, avoiding the oxidation, decomposition and deterioration of the drug, increasing the stability of the drug, and avoiding the use of a large amount of antioxidants, which is in line with national policy orientation and makes clinical drug use safer; 2. Good drug resolubility: the product obtained by freeze drying has a loose texture, and it quickly dissolves and restores the original characteristics of the liquid medicine after adding water; 3. The product dosage is accurate and the appearance is excellent: quantitative infusion after preparing the solution is more accurate than direct packaging of powder; 4. Easy to achieve aseptic operation: the preparation and filling of the liquid medicine are carried out in a sterile clean area, which makes it easier to achieve aseptic production. The liquid medicine passes through a 0.22μm filter ultrafiltration, which can effectively remove bacteria including spores, making the liquid medicine sterile, and can also effectively remove insoluble particles and foreign matter to ensure product quality; 5. The fully automated aseptic production line improves the production environment, avoids harmful dust, and reduces human interference. 6. The production process has good reproducibility and is safer and controllable. However, the existing lyophilized vitamin C powder for injection is difficult to freeze-dry during the preparation process due to the low eutectic point temperature and collapse temperature, resulting in a long preparation process, high energy consumption, cumbersome process, and unstable quality of the prepared lyophilized powder. Therefore, it is necessary to provide a lyophilized vitamin C powder for injection with a simple preparation process, low energy consumption and stable quality. Summary of the invention

[0004] The object of the present invention is to provide a vitamin C freeze-dried powder injection for injection and a preparation method thereof. The present invention uses components such as L-cysteine, tert-butyl alcohol, and β-cyclodextrin to change the eutectic point temperature, glass transition temperature, and collapse temperature of the vitamin C injection, so that the injection is easier to be freeze-dried, the freeze-drying time and the re-dissolution time are significantly shortened, and the vitamin C degradation, discoloration, and content reduction are effectively prevented.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention provides a vitamin C freeze-dried powder injection for injection, which is prepared into 1000 bottles by comprising the following components: 900-1100 g of vitamin C, 650-800 g of excipient, 250-500 g of L-cysteine, 120-250 g of tert-butanol and 300-500 g of beta-cyclodextrin.

[0007] Preferably, the excipient is any one of mannitol, sorbitol, dextran and lactose.

[0008] The present invention also provides a method for preparing the above-mentioned vitamin C freeze-dried powder injection, comprising the following steps:

[0009] (1) Mixing vitamin C, L-cysteine ​​and water to obtain material 1;

[0010] (2) adding β-cyclodextrin to material 1 and subjecting to microwave treatment to obtain material 2;

[0011] (3) adding tert-butyl alcohol and excipients to material 2, adjusting the pH, and filtering to obtain a vitamin C solution;

[0012] (4) freeze-drying the vitamin C solution to obtain the vitamin C freeze-dried powder injection for injection.

[0013] Preferably, the mass of the water is 6 to 10 times the mass of the vitamin C.

[0014] Preferably, the mixing speed is 200 to 500 rpm and the mixing time is 10 to 25 min.

[0015] Preferably, the microwave power is 150-250W and the time is 3-5min.

[0016] Preferably, the pH is adjusted to 5.5-6.5.

[0017] Preferably, the filtration includes activated carbon adsorption and ultrafiltration; the ultrafiltration uses a 0.2-0.3 μm microporous membrane filter.

[0018] Preferably, the freeze-drying procedure includes pre-freezing, sublimation drying, and analytical drying;

[0019] The pre-freezing procedure is: cooling to -15 to -10°C, maintaining for 1 to 2 hours, and further cooling to -30 to -25°C, maintaining for 0.5 to 1 hour, to complete the pre-freezing;

[0020] The sublimation drying procedure is: under a vacuum degree of 0.2 to 0.3 mbar, heating to -20 to -18°C, maintaining for 1 to 2 hours; further heating to -15 to -10°C, maintaining for 1 to 2 hours; further heating to -10 to -5°C, maintaining for 4 to 6 hours; further heating to -5 to 0°C, maintaining for 2 to 3 hours;

[0021] The analytical drying procedure is: under a vacuum degree of 0.1-0.2 mbar, heating to 0-10°C and maintaining for 0.5-1h; further heating to 10-20°C and maintaining for 1-2h; further heating to 35-40°C and maintaining for 2-4h.

[0022] Preferably, during the pre-freezing process, the cooling rate is 10-15°C / min; during the sublimation drying process, the heating rate is 0.5-1.5°C / min; during the analytical drying process, the heating rate is 2-4°C / min.

[0023] The present invention provides a vitamin C freeze-dried powder injection for injection and a preparation method thereof. Compared with the prior art, the present invention has the following beneficial effects:

[0024] (1) The present invention uses tert-butyl alcohol in combination with an excipient, which can increase the eutectic point temperature, glass transition temperature, and collapse temperature of the vitamin C liquid system, thereby increasing the pre-freezing temperature and sublimation temperature during the freeze-drying process, significantly shortening the time required for freeze-drying, saving energy consumption, and simplifying the process.

[0025] (2) The present invention can also improve the molecular structure of the system through microwave treatment, thereby accelerating the re-dissolution speed of the lyophilized vitamin C powder injection.

[0026] (3) The present invention adds L-cysteine ​​to the vitamin C freeze-dried powder for injection. L-cysteine ​​can bind to vitamin C to fix the vitamin C, and further adds β-cyclodextrin to further encapsulate the vitamin C to improve the stability of the vitamin C and reduce the impact of the subsequent preparation process on the activity of the vitamin C, thereby extending the shelf life of the vitamin C freeze-dried powder for injection. DETAILED DESCRIPTION

[0027] The invention provides a vitamin C freeze-dried powder injection for injection, which is prepared into 1000 bottles by comprising the following components: 900-1100 g of vitamin C, 650-800 g of excipient, 250-500 g of L-cysteine, 120-250 g of tert-butanol and 300-500 g of beta-cyclodextrin.

[0028] In the present invention, the lyophilized powder injection is preferably prepared into 1000 bottles by comprising the following components: 900-1100 g of vitamin C, 650-800 g of excipient, 250-500 g of L-cysteine, 120-250 g of tert-butanol, and 300-500 g of β-cyclodextrin; further preferably, it is prepared into 1000 bottles by comprising the following components: 950-1050 g of vitamin C, 700-750 g of excipient, 300-450 g of L-cysteine, 150-200 g of tert-butanol, and 350-450 g of β-cyclodextrin; still further preferably, it is prepared into 1000 bottles by comprising the following components: 1000 g of vitamin C, 750 g of excipient, 400 g of L-cysteine, 180 g of tert-butanol, and 420 g of β-cyclodextrin.

[0029] In the present invention, the excipient is preferably any one of mannitol, sorbitol, dextran and lactose, and more preferably mannitol.

[0030] The present invention also provides a method for preparing the above-mentioned vitamin C freeze-dried powder injection, comprising the following steps:

[0031] (1) Mixing vitamin C, L-cysteine ​​and water to obtain material 1;

[0032] (2) adding β-cyclodextrin to material 1 and subjecting to microwave treatment to obtain material 2;

[0033] (3) adding tert-butyl alcohol and excipients to material 2, adjusting the pH, and filtering to obtain a vitamin C solution;

[0034] (4) freeze-drying the vitamin C solution to obtain the vitamin C freeze-dried powder injection for injection.

[0035] The present invention mixes vitamin C, L-cysteine ​​and water to obtain material 1.

[0036] In the present invention, the mass of the water is preferably 6 to 10 times the mass of the vitamin C, and more preferably 8 times the mass of the vitamin C.

[0037] In the present invention, the water is preferably water for injection.

[0038] In the present invention, the mixing rotation speed is preferably 200 to 500 rpm, and more preferably 300 rpm.

[0039] In the present invention, the mixing time is preferably 10 to 25 minutes, more preferably 20 minutes.

[0040] The present invention adds beta-cyclodextrin to material 1 and performs microwave treatment to obtain material 2.

[0041] In the present invention, the power of the microwave is preferably 150-250W, more preferably 200W.

[0042] In the present invention, the microwave time is preferably 3 to 5 minutes, more preferably 4 minutes.

[0043] The present invention adds tert-butyl alcohol and excipients to material 2, adjusts the pH, and obtains vitamin C injection through filtration.

[0044] In the present invention, the pH is preferably adjusted to 5.5 to 6.5, and more preferably adjusted to 6.0.

[0045] In the present invention, the pH adjusting agent is preferably sodium hydroxide.

[0046] In the present invention, the filtration preferably includes activated carbon adsorption and ultrafiltration.

[0047] In the present invention, the method of activated carbon adsorption is preferably to use 0.02-0.1% (w / v) activated carbon (for injection), adsorb for 10-20 minutes, and then filter to remove carbon; further preferably to use 0.02% (w / v) activated carbon (for injection), adsorb for 15 minutes, and then filter to remove carbon.

[0048] In the present invention, the ultrafiltration preferably uses a 0.2-0.3 μm microporous membrane filter, and more preferably uses a 0.22 μm microporous membrane filter.

[0049] The present invention freeze-dries the vitamin C medicinal solution to obtain the vitamin C freeze-dried powder injection for injection.

[0050] In the present invention, the freeze-drying procedure preferably includes pre-freezing, sublimation drying, and analytical drying.

[0051] In the present invention, the pre-freezing procedure is preferably: cooling to -15 to -10°C, maintaining for 1 to 2 hours, continuing to cool to -30°C to -25°C, maintaining for 0.5 to 1 hour, and completing the pre-freezing; further preferably, cooling to -15°C, maintaining for 1.5 hours, continuing to cool to -30°C, maintaining for 0.5 hour, and completing the pre-freezing.

[0052] In the present invention, the cooling rate is preferably 10-15°C / min, more preferably 12°C / min.

[0053] In the present invention, the sublimation drying procedure is preferably: under a vacuum degree of 0.2 to 0.3 mbar, heating to -20 to -18 ° C, maintaining for 1 to 2 hours, continuing to heat to -15 to -10 ° C, maintaining for 1 to 2 hours, continuing to heat to -10 to -5 ° C, maintaining for 4 to 6 hours, continuing to heat to -5 to 0 ° C, maintaining for 2 to 3 hours; further preferably, under a vacuum degree of 0.25 mbar, heating to -20 ° C, maintaining for 2 hours, continuing to heat to -10 ° C, maintaining for 1 hour, continuing to heat to -5 ° C, maintaining for 5 hours, continuing to heat to 0 ° C, maintaining for 2.5 hours.

[0054] In the present invention, the heating rate is preferably 0.5-1.5°C / min, more preferably 1.0°C / min.

[0055] In the present invention, the analytical drying procedure is preferably: under a vacuum degree of 0.1 to 0.2 mbar, heating to 0 to 10°C, maintaining for 0.5 to 1 h, continuing to heat to 10 to 20°C, maintaining for 1 to 2 h, continuing to heat to 35 to 40°C, maintaining for 2 to 4 h; further preferably, under a vacuum degree of 0.1 mbar, heating to 5°C, maintaining for 0.5 h, continuing to heat to 15°C, maintaining for 2 h, continuing to heat to 40°C, maintaining for 3 h.

[0056] In the present invention, the heating rate is preferably 2-4°C / min, more preferably 3°C / min.

[0057] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0058] Example 1

[0059] This embodiment provides a vitamin C freeze-dried powder injection for injection, and the specific preparation process is as follows:

[0060] (1) According to the dosage of 1000 bottles, prepare 1000g of vitamin C, 750g of mannitol, 400g of L-cysteine, 180g of tert-butyl alcohol, and 420g of β-cyclodextrin and set aside.

[0061] (2) Mix vitamin C, L-cysteine ​​and 8000 mL of water for injection, and stir at 300 rpm for 20 min to obtain material 1.

[0062] (3) Add β-cyclodextrin to material 1, stir evenly, and treat with a 200 W microwave for 4 min to obtain material 2.

[0063] (4) Add tert-butyl alcohol and mannitol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution.

[0064] (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0065] Cool down to -15°C at a rate of 12°C / min, keep for 1.5h, then continue to cool down to -30°C, keep for 0.5h, and complete pre-freezing;

[0066] Adjust the vacuum degree to 0.25 mbar, and heat at a rate of 1.0 °C / min, first heat to -20 °C, hold for 2 h, then heat to -10 °C, hold for 1 h, then heat to -5 °C, hold for 5 h, then heat to 0 °C, hold for 2.5 h, and complete sublimation drying;

[0067] The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 3°C / min, first to 5°C, maintained for 0.5 h, then to 15°C, maintained for 2 h, then to 40°C, maintained for 3 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

[0068] The obtained freeze-dried preparation has a full and fluffy appearance and good formability.

[0069] Example 2

[0070] This embodiment provides a vitamin C freeze-dried powder injection for injection, and the specific preparation process is as follows:

[0071] (1) Prepare 900 g of vitamin C, 650 g of mannitol, 250 g of L-cysteine, 120 g of tert-butyl alcohol, and 300 g of β-cyclodextrin according to the dosage of 1000 bottles and set aside.

[0072] (2) Vitamin C, L-cysteine ​​and 5400 mL of water for injection were mixed and stirred at 200 rpm for 20 min to obtain material 1.

[0073] (3) Add β-cyclodextrin to material 1, stir evenly, and treat with a 250W microwave for 3 minutes to obtain material 2.

[0074] (4) Add tert-butyl alcohol and mannitol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution.

[0075] (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0076] Cool down to -15°C at a rate of 10°C / min, keep for 1 hour, and continue to cool down to -30°C, keep for 1 hour to complete pre-freezing;

[0077] Adjust the vacuum degree to 0.2 mbar, and heat at a rate of 0.5 °C / min, first heat to -20 °C, hold for 1 h, then heat to -10 °C, hold for 2 h, then heat to -5 °C, hold for 6 h, then heat to 0 °C, hold for 2 h, and complete sublimation drying;

[0078] The vacuum degree was adjusted to 0.15 mbar, and the temperature was increased at a rate of 2°C / min, first to 5°C, maintained for 1 h, then to 15°C, maintained for 1 h, then to 40°C, maintained for 4 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

[0079] The obtained freeze-dried preparation has a full and fluffy appearance and good formability.

[0080] Example 3

[0081] This embodiment provides a vitamin C freeze-dried powder injection for injection, and the specific preparation process is as follows:

[0082] (1) Prepare 1100 g of vitamin C, 800 g of mannitol, 500 g of L-cysteine, 250 g of tert-butyl alcohol, and 500 g of β-cyclodextrin according to the dosage of 1000 bottles and set aside.

[0083] (2) Mix vitamin C and L-cysteine ​​with 10000 mL of water for injection and stir at 500 rpm for 20 min to obtain material 1.

[0084] (3) Add β-cyclodextrin to material 1, stir evenly, and treat with a 150 W microwave for 5 min to obtain material 2.

[0085] (4) Add tert-butyl alcohol and mannitol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution.

[0086] (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0087] Cool down to -15°C at a rate of 15°C / min, keep for 1.5h, continue to cool down to -25°C, keep for 1h, and complete pre-freezing;

[0088] Adjust the vacuum degree to 0.3 mbar, and heat at a rate of 1.5 °C / min, first heat to -20 °C, hold for 1 h, then heat to -15 °C, hold for 1 h, then heat to -10 °C, hold for 5 h, then heat to -5 °C, hold for 2.5 h, and complete sublimation drying;

[0089] The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 4°C / min, first to 10°C and maintained for 0.5 h, then to 20°C and maintained for 2 h, then to 35°C and maintained for 3 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

[0090] The obtained freeze-dried preparation has a full and fluffy appearance and good formability.

[0091] Comparative Example 1

[0092] The difference between this comparative example and Example 1 is that L-cysteine ​​is omitted. The specific process is as follows:

[0093] (1) Prepare 1000 g of vitamin C, 750 g of mannitol, 180 g of tert-butyl alcohol, and 420 g of β-cyclodextrin according to the dosage of 1000 bottles and set aside.

[0094] (2) Mix vitamin C with 8000 mL of water for injection and stir at 300 rpm for 20 min to obtain material 1.

[0095] (3) Add β-cyclodextrin to material 1, stir evenly, and treat with a 200 W microwave for 4 min to obtain material 2.

[0096] (4) Add tert-butyl alcohol and mannitol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution.

[0097] (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0098] Cool down to -15°C at a rate of 12°C / min, keep for 1.5h, then continue to cool down to -30°C, keep for 0.5h, and complete pre-freezing;

[0099] Adjust the vacuum degree to 0.25 mbar, and heat at a rate of 1.0 °C / min, first heat to -20 °C, hold for 2 h, then heat to -10 °C, hold for 1 h, then heat to -5 °C, hold for 5 h, then heat to 0 °C, hold for 2.5 h, and complete sublimation drying;

[0100] The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 3°C / min, first to 5°C, maintained for 0.5 h, then to 15°C, maintained for 2 h, then to 40°C, maintained for 3 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

[0101] The obtained freeze-dried preparation has a full and fluffy appearance and good formability.

[0102] Comparative Example 2

[0103] The difference between this comparative example and Example 1 is that the excipient is omitted. The specific process is as follows:

[0104] (1) Prepare 1000 g of vitamin C, 400 g of L-cysteine, 180 g of tert-butyl alcohol, and 420 g of β-cyclodextrin according to the dosage of 1000 bottles and set aside.

[0105] (2) Mix vitamin C, L-cysteine ​​and 8000 mL of water for injection, and stir at 300 rpm for 20 min to obtain material 1.

[0106] (3) Add β-cyclodextrin to material 1, stir evenly, and treat with a 200 W microwave for 4 min to obtain material 2.

[0107] (4) Add tert-butyl alcohol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution.

[0108] (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0109] Cool down to -15°C at a rate of 12°C / min, keep for 1.5h, then continue to cool down to -30°C, keep for 0.5h, and complete pre-freezing;

[0110] Adjust the vacuum degree to 0.25 mbar, and heat at a rate of 1.0 °C / min, first heat to -20 °C, hold for 2 h, then heat to -10 °C, hold for 1 h, then heat to -5 °C, hold for 5 h, then heat to 0 °C, hold for 2.5 h, and complete sublimation drying;

[0111] The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 3°C / min, first to 5°C, maintained for 0.5 h, then to 15°C, maintained for 2 h, then to 40°C, maintained for 3 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

[0112] The obtained lyophilized agent has an incomplete appearance, is collapsed, and has residual crystals.

[0113] Comparative Example 3

[0114] The difference between this comparative example and Example 1 is that tert-butyl alcohol is omitted. The specific process is as follows:

[0115] (1) Prepare 1000 g of vitamin C, 750 g of mannitol, 400 g of L-cysteine, and 420 g of β-cyclodextrin according to the dosage of 1000 bottles and set aside.

[0116] (2) Mix vitamin C, L-cysteine ​​and 8000 mL of water for injection, and stir at 300 rpm for 20 min to obtain material 1.

[0117] (3) Add β-cyclodextrin to material 1, stir evenly, and treat with a 200 W microwave for 4 min to obtain material 2.

[0118] (4) Add mannitol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution.

[0119] (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0120] Cool down to -15°C at a rate of 12°C / min, keep for 1.5h, then continue to cool down to -30°C, keep for 0.5h, and complete pre-freezing;

[0121] Adjust the vacuum degree to 0.25 mbar, and heat at a rate of 1.0 °C / min, first heat to -20 °C, hold for 2 h, then heat to -10 °C, hold for 1 h, then heat to -5 °C, hold for 5 h, then heat to 0 °C, hold for 2.5 h, and complete sublimation drying;

[0122] The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 3°C / min, first to 5°C, maintained for 0.5 h, then to 15°C, maintained for 2 h, then to 40°C, maintained for 3 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

[0123] The obtained lyophilized agent has an incomplete appearance, is collapsed, and has residual crystals.

[0124] Comparative Example 4

[0125] The difference between this comparative example and Example 1 is that β-cyclodextrin is omitted. The specific process is as follows:

[0126] (1) Prepare 1000 g of vitamin C, 750 g of mannitol, 400 g of L-cysteine, and 180 g of tert-butyl alcohol according to the dosage of 1000 bottles and set aside.

[0127] (2) Vitamin C, L-cysteine ​​and 8000 mL of water for injection were mixed and stirred at 300 rpm for 20 min to obtain material 1. Material 1 was treated with a 200 W microwave for 4 min to obtain material 2.

[0128] (3) Add tert-butyl alcohol and mannitol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution.

[0129] (4) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0130] Cool down to -15°C at a rate of 12°C / min, keep for 1.5h, then continue to cool down to -30°C, keep for 0.5h, and complete pre-freezing;

[0131] Adjust the vacuum degree to 0.25 mbar, and heat at a rate of 1.0 °C / min, first heat to -20 °C, hold for 2 h, then heat to -10 °C, hold for 1 h, then heat to -5 °C, hold for 5 h, then heat to 0 °C, hold for 2.5 h, and complete sublimation drying;

[0132] The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 3°C / min, first to 5°C, maintained for 0.5 h, then to 15°C, maintained for 2 h, then to 40°C, maintained for 3 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

[0133] The obtained lyophilized agent has an incomplete appearance, is collapsed, and has residual crystals.

[0134] Comparative Example 5

[0135] The difference between this comparative example and Example 1 is that the microwave treatment is omitted. The specific process is as follows:

[0136] This embodiment provides a vitamin C freeze-dried powder injection for injection, and the specific preparation process is as follows:

[0137] (1) According to the dosage of 1000 bottles, prepare 1000g of vitamin C, 750g of mannitol, 400g of L-cysteine, 180g of tert-butyl alcohol, and 420g of β-cyclodextrin and set aside.

[0138] (2) Mix vitamin C, L-cysteine ​​and 8000 mL of water for injection, and stir at 300 rpm for 20 min to obtain material 1.

[0139] (3) Add β-cyclodextrin to material 1 and stir evenly to obtain material 2.

[0140] (4) Add tert-butyl alcohol and mannitol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution.

[0141] (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0142] Cool down to -15°C at a rate of 12°C / min, keep for 1.5h, then continue to cool down to -30°C, keep for 0.5h, and complete pre-freezing;

[0143] Adjust the vacuum degree to 0.25 mbar, and heat at a rate of 1.0 °C / min, first heat to -20 °C, hold for 2 h, then heat to -10 °C, hold for 1 h, then heat to -5 °C, hold for 5 h, then heat to 0 °C, hold for 2.5 h, and complete sublimation drying;

[0144] The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 3°C / min, first to 5°C, maintained for 0.5 h, then to 15°C, maintained for 2 h, then to 40°C, maintained for 3 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

[0145] The obtained lyophilized agent has an incomplete appearance, is collapsed, and has residual crystals.

[0146] Comparative Example 6

[0147] The difference between this comparative example and Example 1 is that the freeze-drying procedure is adjusted. The specific process is as follows:

[0148] This embodiment provides a vitamin C freeze-dried powder injection for injection, and the specific preparation process is as follows:

[0149] (1) Prepare 1000 g of vitamin C, 750 g of mannitol, 400 g of L-cysteine, 180 g of tert-butyl alcohol, and 420 g of β-cyclodextrin according to the dosage of 1000 bottles and set aside.

[0150] (2) Mix vitamin C, L-cysteine ​​and 8000 mL of water for injection, and stir at 300 rpm for 20 min to obtain material 1.

[0151] (3) Add β-cyclodextrin to material 1, stir evenly, and treat with a 200 W microwave for 4 min to obtain material 2.

[0152] (4) Add tert-butyl alcohol and mannitol to material 2, stir evenly, and adjust the pH to 6.0 with 0.1M sodium hydroxide. Add 0.02% (w / v) activated carbon (for injection) to the solution after pH adjustment, stir and adsorb for 15 minutes, and then filter to remove carbon. Then filter and sterilize the solution with a 0.22μm microporous membrane filter to obtain a vitamin C solution. The resulting lyophilized agent

[0153] (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying:

[0154] Cool down to -30°C at a rate of 5°C / min and maintain for 6 hours to complete pre-freezing;

[0155] Adjust the vacuum degree to 0.25 mbar, and heat at a rate of 2.0 °C / min, first heat to -20 °C, hold for 2 h, then heat to -10 °C, hold for 4 h, then heat to -5 °C, hold for 5 h, then heat to 0 °C, hold for 3 h, and complete sublimation drying;

[0156] The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 5°C / min, first to 10°C and maintained for 2.5 h, then to 40°C and maintained for 6 h to complete the desorption and drying to obtain the lyophilized vitamin C powder injection.

[0157] The obtained freeze-dried preparation has a stratified appearance, and the lower layer is shrunken.

[0158] Test Example 1

[0159] This test example tested the relevant indicators of the vitamin C freeze-dried powder for injection prepared in Examples 1 to 3 and Comparative Examples 1 to 6. The specific process is as follows:

[0160] 1. Safety test:

[0161] 1.1 Intravenous vascular irritation test

[0162] 20 healthy rabbits weighing 2.0-2.5kg were selected and randomly divided into 10 groups, with 2 rabbits in each group. Among them, 9 groups were injected with the vitamin C solution for injection prepared by Examples 1-3 and Comparative Examples 1-6 into the right ear vein of the rabbits as the experimental group, and 1 group was injected with an equal amount of normal saline as the negative control group. The vitamin C solution for injection has a mass concentration of 10.0g / L (i.e., 1L solution contains VC 10.0g) according to vitamin C, and the injection volume is 1.0mL / kg body weight, once a day for 3 consecutive days. After administration, the local blood vessels of the administration were observed with the naked eye for thrombosis, and the surrounding tissues were observed for inflammatory reactions such as redness, swelling, and congestion. 96h after the last administration, the proximal and distal ends of the injection site were observed for erythema and edema.

[0163] result:

[0164] During the administration period, no thrombosis, redness, swelling or congestion was observed in the auricular vein of rabbits in each group, and no significant difference was observed between the treated side and the untreated side. After the last administration, no obvious erythema or edema was observed at the proximal and distal ends of the injection site of rabbits in each group.

[0165] 1.2 Hemolytic test

[0166] Take about 20 mL of rabbit blood, put it in a beaker, remove fibrinogen, add about 10 times the amount of physiological saline, shake well, centrifuge, remove the supernatant, and wash the precipitated red blood cells with physiological saline for 3 times until the supernatant is no longer red. Dilute the obtained red blood cells with physiological saline to a suspension with a volume fraction of 2%.

[0167] The vitamin C freeze-dried powder for injection prepared in Examples 1 to 3 and Comparative Examples 1 to 6 was prepared into a 10.0 g / L solution with water for injection as a test sample, and the following experiments were performed respectively:

[0168] According to the dosage in the reaction system shown in Table 1, add 2% red blood cell suspension and physiological saline in turn, mix well, place in a 37℃ constant temperature water bath for 0.5h, then add different amounts of test samples, tube 1 is the negative control tube, tube 7 is the positive control tube, shake well, place in a 37℃ constant temperature box, observe once every 15min at first, observe once every 1h after 1h, and observe continuously for 4h. If the solution is transparent red, it means hemolysis. If there is brown-red or reddish-brown flocculent precipitation in the solution, it means red blood cell hemolytic reaction.

[0169] Table 1 Reaction system

[0170]

[0171]

[0172] result:

[0173] During the hemolytic test of the lyophilized vitamin C powder for injection in each group, only tube 7 showed hemolysis, while tubes 1 to 6 showed no hemolysis or agglutination.

[0174] The above test results show that the lyophilized vitamin C powder for injection prepared in Examples 1 to 3 and Comparative Examples 1 to 6 are safe and can be used for clinical application.

[0175] 2. Determination of re-dissolution time

[0176] Three bottles of lyophilized vitamin C powder for injection prepared in Examples 1 to 3 and Comparative Examples 1 to 6 were taken, 10 mL of water for injection was added, and the dissolution time of the lyophilized powder for injection was calculated and the average value was taken. The results are shown in Table 2.

[0177] Table 2 Reconstitution time of each group of vitamin C lyophilized powder injection

[0178] Group Reconstitution time Example 1 4 minutes 12 seconds Example 2 4 minutes 18 seconds Example 3 4 minutes 15 seconds Comparative Example 1 4 minutes 32 seconds Comparative Example 2 5 minutes and 46 seconds Comparative Example 3 4 minutes 53 seconds Comparative Example 4 7 minutes 33 seconds Comparative Example 5 6 minutes 11 seconds Comparative Example 6 6 minutes 37 seconds

[0179] According to Table 2, the reconstitution time of the lyophilized vitamin C powder for injection prepared in Examples 1 to 3 and Comparative Examples 1 to 6 is within 5 minutes, and the dissolution rate is relatively fast. The reconstitution time of Comparative Examples 1 and 3 is also within 5 minutes, which indicates that L-cysteine ​​and tert-butyl alcohol have little effect on the reconstitution time of the lyophilized agent; while the reconstitution time of Comparative Examples 2, 3, 4, and 5 exceeds 5 minutes, which indicates that excipients, β-cyclodextrin, microwave treatment, and inappropriate lyophilization procedures have an effect on the reconstitution time of the lyophilized agent, especially the use of β-cyclodextrin, which has a significant effect on the reconstitution time.

[0180] 3. Vitamin C stability determination

[0181] The lyophilized powders of vitamin C for injection prepared in Examples 1 to 3 and Comparative Examples 1 to 6 were placed at 25 ± 2 ° C for 24 months, and samples were taken at the 0th, 3rd, 6th, 12th and 24th months. Referring to the content determination method of Part II of the 2020 edition of the Chinese Pharmacopoeia, each group of lyophilized powders of vitamin C for injection was accurately weighed, 0.2 g of vitamin C was calculated, 100 ml of freshly boiled cold water and 10 ml of dilute acetic acid were added to dissolve it, 1 ml of starch indicator solution was added, and iodine titrant (0.05 mol / L) was immediately titrated until the solution turned blue and did not fade within 30 seconds. Each 1 ml of iodine titrant (0.05 mol / L) is equivalent to 8.806 mg of C6H8O6.

[0182] The standard stipulates that based on the average packaging amount, the vitamin C content should be 95.0-105.0% of the labeled amount.

[0183] The results are shown in Table 3.

[0184] Table 3 Stability test results of each group of vitamin C lyophilized powder injection

[0185]

[0186] According to Table 3, it can be seen that the content of vitamin C in the vitamin C freeze-dried powder for injection prepared in Examples 1 to 3 is relatively stable, and no significant changes occurred within 24 months. The vitamin C content in Comparative Examples 1, 4, and 5 has been significantly reduced, which shows that L-cysteine, β-cyclodextrin and microwave treatment also have a significant effect on the stability of vitamin C, especially the use of L-cysteine ​​and β-cyclodextrin, which has the greatest effect on the stability of vitamin C. The vitamin C content in Comparative Examples 2, 3, and 5 has also decreased, but the degree is low and still meets the standard.

[0187] Test Example 2

[0188] In this test example, the collapse temperature, glass transition temperature and eutectic point temperature of the vitamin C solution in the preparation process of Examples 1 to 3 and Comparative Examples 1 to 5 were measured (the solution system of Comparative Example 6 was consistent with that of Examples 1 to 3, therefore, Comparative Example 6 was not tested). The results are shown in Table 4.

[0189] Table 4 Collapse temperature, glass transition temperature and eutectic point temperature of each group of vitamin C solution

[0190] Group Collapse temperature(℃) Glass transition temperature(℃) Eutectic temperature(℃) Example 1 -27.35 -30.1 -21.51 Example 2 -27.32 -30.3 -21.47 Example 3 -27.36 -30.1 -21.50 Comparative Example 1 -27.33 -30.8 -21.59 Comparative Example 2 -30.69 -37.5 -24.56 Comparative Example 3 -31.51 -36.7 -23.46 Comparative Example 4 -29.41 -32.5 -22.60 Comparative Example 5 -27.35 -30.1 -21.51

[0191] From the above test results, it can be seen that the collapse temperature, glass transition temperature and eutectic point temperature of the drug solution of Examples 1 to 3 are higher than those of Comparative Examples 1 to 5, which makes it easier for the drug solution to crystallize and sublimate during the freeze-drying process, and the water is easier to remove, thereby shortening the freeze-drying time. When the injection solution of Comparative Examples 1 to 5 adopts the freeze-drying procedure of Examples 1 to 3, the formation of the freeze-dried powder injection is not as good as that of Examples 1 to 3, and when the drug solution of Comparative Example 6 with the same system as the embodiment adopts a different freeze-drying procedure, its freeze-drying effect is also not as good as that of Examples 1 to 3. It can be seen that the drug solution system composition and freeze-drying procedure of the present invention need to be adopted to ensure the freeze-drying effect and stability effect of the freeze-dried powder injection of vitamin C for injection.

[0192] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A vitamin C freeze-dried powder injection, characterized in that: 1000 bottles are prepared by including the following components: 1000 g of vitamin C, 750 g of mannitol, 400 g of L-cysteine, 180 g of tert-butyl alcohol, and 420 g of β-cyclodextrin; The preparation method of the vitamin C freeze-dried powder injection for injection comprises the following steps: (1) According to the dosage of 1000 bottles, prepare 1000g of vitamin C, 750g of mannitol, 400g of L-cysteine, 180g of tert-butyl alcohol, and 420g of β-cyclodextrin for use. (2) Mix vitamin C, L-cysteine ​​and 8000 mL of water for injection, and stir at 300 rpm for 20 min to obtain material 1. (3) Add β-cyclodextrin to material 1, stir evenly, and treat with 200W microwave for 4min to obtain material 2. (4) tert-butyl alcohol and mannitol were added to material 2, and after stirring evenly, the pH was adjusted to 6.0 with 0.1 M sodium hydroxide, and 0.02% (w / v) activated carbon was added to the solution after pH adjustment. After stirring and adsorbing for 15 minutes, the solution was filtered to remove carbon, and then the solution was filtered and sterilized with a 0.22 μm microporous membrane filter to obtain a vitamin C solution. (5) The vitamin C solution was packaged and placed in a freeze dryer, and the following parameters were set for freeze drying: Cool down to -15°C at a rate of 12°C / min, keep for 1.5h, then continue to cool down to -30°C, keep for 0.5h, and complete pre-freezing; Adjust the vacuum degree to 0.25 mbar, and heat at a rate of 1.0 °C / min, first heat to -20 °C, hold for 2 h, then heat to -10 °C, hold for 1 h, then heat to -5 °C, hold for 5 h, then heat to 0 °C, hold for 2.5 h, and complete sublimation drying; The vacuum degree was adjusted to 0.1 mbar, and the temperature was increased at a rate of 3°C / min, first to 5°C, maintained for 0.5 h, then to 15°C, maintained for 2 h, then to 40°C, maintained for 3 h, to complete the desorption and drying, and obtain the lyophilized vitamin C powder for injection.

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