Vitamin B12 injection and preparation method thereof

Abstract

The invention discloses a vitamin B12 injection and a preparation method thereof, and relates to the technical field of pharmaceutical preparations. The preparation method of the injection comprises the following steps: (1) dissolving the raw material medicines: dissolving the vitamin B12 and the benzyl alcohol in the water for injection to obtain an injection A; (2) dissolving a stabilizer-synergist: firstly adding L-cysteine into the injection A, then adding liraglutide, and uniformly mixing to obtain an injection B; (3) pH value adjustment: adjusting the pH value of the injection B to 4.0-4.5 to obtain an injection C; (4) dissolution of an impurity reducing agent-osmotic pressure regulator: sequentially adding spleen aminopeptide and glucose into the injection C, and standing to obtain an injection D; (5) fixing the volume, filtering and sterilizing; and (6) filling. According to the vitamin B12 injection prepared by the method, the preparation time can be greatly shortened, and total impurities in a final finished product can be obviously reduced through reasonable dissolution sequence, selection of stirring materials and limitation of temperature and pH value.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and in particular to a vitamin B12 injection and a preparation method thereof. Background Art

[0002] Vitamin B12 is also known as cobalamin. Its chemical name is Coα-[α-(5,6-dimethylbenzimidazolyl)]-Coβ cyanocobalamide. It is a type of B vitamin composed of cobalt-containing porphyrin compounds. It is the only vitamin containing metal elements. It is dark red or red crystalline powder. It is easily soluble in water and ethanol. It is most stable under weak acid conditions of pH 4.5-5.0. It decomposes in strong acid or alkaline solutions. It can be damaged to a certain extent when exposed to heat. It is easily damaged when exposed to strong light or ultraviolet rays. It is odorless, hygroscopic, and easily absorbed by the human body. Vitamin B12 can participate in the synthesis of DNA as a cofactor of methyltransferase, promote the development and maturation of red blood cells, maintain the normal hematopoietic function of the body, prevent pernicious anemia, and maintain the health of the nervous system. In clinical practice, it is mostly administered by injection. Injectable vitamin B12 can be quickly absorbed by the body.

[0003] At present, the preparation process of existing vitamin B12 injection includes dissolution of raw materials, dissolution of osmotic pressure regulator, solution pH dissolution, constant volume, filtration and sterilization, and filling to obtain vitamin B12 injection. However, there are still many problems in the existing process, such as the rapid dissolution of vitamin B12 raw materials cannot be well achieved, thereby causing the vitamin B12 raw materials to degrade during the preparation process, resulting in high impurity content and variety of prepared vitamin B12 injection, and obvious impurity growth during storage. Therefore, developing a vitamin B12 injection with low impurity content and high storage stability is of great significance to improving the medication safety of patients. Summary of the invention

[0004] In view of the above-mentioned defects of the prior art, the present invention proposes a vitamin B12 injection and a preparation method thereof to solve the problems raised by the above-mentioned background technology.

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

[0006] A vitamin B12 injection, each mL of the injection contains the following ingredients: 0.8-1.2 mg of vitamin B12, 10-15 mg of liraglutide, 6-12 mg of spleen aminopeptide, 3-7 mg of L-cysteine, 0.5-5 mg of glucose, 0.3-1 mg of benzyl alcohol, 1-10 mg of a pH regulator, and the balance is water for injection.

[0007] Preferably, each mL of injection contains the following ingredients: 1.0 mg vitamin B12, 12 mg liraglutide, 8 mg spleen aminopeptide, 6 mg L-cysteine, 1 mg glucose, 0.8 mg benzyl alcohol, 5 mg pH regulator, and the balance is water for injection.

[0008] Preferably, the preparation method of vitamin B12 injection comprises the following steps:

[0009] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them to water for injection at a temperature of 30-35°C, stir and dissolve to obtain injection solution A;

[0010] (2) Dissolving the stabilizer-enhancer: first add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix, then add liraglutide, mix and mix to obtain injection solution B;

[0011] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.0-4.5 to obtain injection solution C;

[0012] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix, then add glucose, and let stand for 20 to 30 minutes to obtain D injection;

[0013] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection and filtered for sterilization;

[0014] (6) Filling: Fill, seal and sterilize the filtered and sterilized injection solution according to the specifications to obtain the vitamin B12 injection solution.

[0015] Preferably, the preparation method is carried out under light-proof conditions.

[0016] Preferably, the temperature of the water for injection in step (1) is 32°C.

[0017] Preferably, the stirring rod used for stirring is made of titanium.

[0018] Preferably, the pH adjusting agent in step (3) is 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution.

[0019] Preferably, in step (3), the pH value is adjusted to 4.2.

[0020] Preferably, in step (5), filtration is performed using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane in sequence.

[0021] Preferably, the sterilization process parameters in step (6) are a sterilization temperature of 120°C and a sterilization time of 10 to 13 minutes.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] (1) The present invention can reduce the overall dissolution time of vitamin B12 by controlling the dissolution order of each component and the temperature of the injection water during the preparation of the injection, thereby greatly reducing the preparation time of the vitamin B12 injection, thereby reducing the production cost and reducing the risk of exposure to degradation and contamination of the drug solution due to excessive preparation time, thereby ensuring a low degradation failure rate of vitamin B12;

[0024] (2) The stirring rod material selected in the present invention is titanium, which does not react with vitamin B12 and does not cause the formation of impurities in the injection, thereby effectively maintaining the stability of the vitamin B12 injection;

[0025] (3) The method of the present invention does not carry out nitrogen filling protection during the preparation process, which is different from the prior art. At the same time, the present invention utilizes the mutual matching reaction of liraglutide, spleen aminopeptide, L-cysteine ​​and glucose to give full play to the synergistic effect of liraglutide, the total impurity reduction effect of spleen aminopeptide, the stabilizing effect of L-cysteine ​​and the osmotic pressure regulating effect of glucose, and the vitamin B12 injection finally obtained is non-irritating, highly stable, not easy to oxidize, has less impurity increment in the sterilization process, does not have the problem of drug solution adhesion, and is easy to operate, so that the prepared vitamin B6 injection has good stability. DETAILED DESCRIPTION

[0026] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0027] Example 1

[0028] A vitamin B12 injection, each mL of injection contains the following ingredients: 0.8 mg of vitamin B12, 10 mg of liraglutide, 6 mg of spleen aminopeptide, 3 mg of L-cysteine, 0.5 mg of glucose, 0.3 mg of benzyl alcohol, 1 mg of pH regulator, and the balance is water for injection.

[0029] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0030] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 30°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0031] (2) Dissolution of stabilizer-enhancer: First, add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix using a titanium rod stirring rod, then add liraglutide and mix to obtain injection solution B;

[0032] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.0 to obtain injection solution C;

[0033] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, then add glucose, and let stand for 20 minutes to obtain D injection;

[0034] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane;

[0035] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it and sterilize it at 120°C for 10 min to obtain vitamin B12 injection solution.

[0036] Example 2

[0037] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.2 mg of vitamin B12, 15 mg of liraglutide, 12 mg of spleen aminopeptide, 7 mg of L-cysteine, 5 mg of glucose, 1 mg of benzyl alcohol, 10 mg of pH regulator, and the balance is water for injection.

[0038] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0039] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 35°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0040] (2) Dissolution of stabilizer-enhancer: First, add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix using a titanium stirring rod, then add liraglutide and mix to obtain injection solution B;

[0041] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.5 to obtain injection solution C;

[0042] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, then add glucose, and let stand for 30 minutes to obtain D injection;

[0043] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane;

[0044] (6) Filling: Fill the filtered and sterilized injection solution into a medium borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 13 min to obtain the vitamin B12 injection solution.

[0045] Example 3

[0046] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 12 mg of liraglutide, 8 mg of spleen aminopeptide, 6 mg of L-cysteine, 1 mg of glucose, 0.8 mg of benzyl alcohol, 5 mg of pH regulator, and the balance is water for injection.

[0047] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0048] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 32°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0049] (2) Dissolution of stabilizer-enhancer: First, add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix using a titanium rod stirring rod, then add liraglutide and mix to obtain injection solution B;

[0050] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.2 to obtain injection solution C;

[0051] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, then add glucose, and let stand for 25 minutes to obtain D injection;

[0052] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane in sequence;

[0053] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 12 min to obtain vitamin B12 injection solution.

[0054] Comparative Example 1

[0055] This comparative example is different from Example 3 in that liraglutide is not added to the ingredients, and the other ingredients and steps remain unchanged.

[0056] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 8 mg of spleen aminopeptide, 6 mg of L-cysteine, 1 mg of glucose, 0.8 mg of benzyl alcohol, 5 mg of pH regulator, and the balance is water for injection.

[0057] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0058] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 32°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0059] (2) Dissolution of stabilizer-enhancer: L-cysteine ​​was first added to the injection solution A obtained in step (1), and the mixture was stirred with a titanium stirring rod to obtain injection solution B;

[0060] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.2 to obtain injection solution C;

[0061] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, then add glucose, and let stand for 25 minutes to obtain D injection;

[0062] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane in sequence;

[0063] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 12 min to obtain vitamin B12 injection solution.

[0064] Comparative Example 2

[0065] This comparative example is different from Example 3 in that liraglutide is replaced with glutathione, and the other ingredients and steps remain unchanged.

[0066] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 12 mg of glutathione, 8 mg of spleen aminopeptide, 6 mg of L-cysteine, 1 mg of glucose, 0.8 mg of benzyl alcohol, 5 mg of pH regulator, and the balance is water for injection.

[0067] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0068] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 32°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0069] (2) Dissolution of stabilizer-enhancer: First, add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix using a titanium rod stirring rod, then add glutathione and mix to obtain injection solution B;

[0070] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.2 to obtain injection solution C;

[0071] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, then add glucose, and let stand for 25 minutes to obtain D injection;

[0072] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane in sequence;

[0073] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 12 min to obtain vitamin B12 injection solution.

[0074] Comparative Example 3

[0075] This comparative example is different from Example 3 in that spleen aminopeptide is not added to the ingredients, and the other ingredients and steps remain unchanged.

[0076] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 12 mg of liraglutide, 6 mg of L-cysteine, 1 mg of glucose, 0.8 mg of benzyl alcohol, 5 mg of a pH adjuster, and the balance is water for injection.

[0077] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0078] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 32°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0079] (2) Dissolution of stabilizer-enhancer: First, add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix using a titanium rod stirring rod, then add liraglutide and mix to obtain injection solution B;

[0080] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.2 to obtain injection solution C;

[0081] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: Add glucose to the injection solution C obtained in step (3) and let stand for 25 minutes to obtain injection solution D;

[0082] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane in sequence;

[0083] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 12 min to obtain vitamin B12 injection solution.

[0084] Comparative Example 4

[0085] This comparative example differs from Example 3 in that L-cysteine ​​is not added to the ingredients, and the other ingredients and steps remain unchanged.

[0086] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 12 mg of liraglutide, 8 mg of spleen aminopeptide, 1 mg of glucose, 0.8 mg of benzyl alcohol, 5 mg of pH regulator, and the balance is water for injection.

[0087] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0088] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 32°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0089] (2) Dissolution of stabilizer-enhancer: adding liraglutide to the injection solution A obtained in step (1) to obtain injection solution B;

[0090] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.2 to obtain injection solution C;

[0091] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, then add glucose, and let stand for 25 minutes to obtain D injection;

[0092] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane in sequence;

[0093] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 12 min to obtain vitamin B12 injection solution.

[0094] Comparative Example 5

[0095] The difference between this comparative example and Example 3 is that L-cysteine ​​in the ingredients is replaced with EDTA-2Na, and the other ingredients and steps remain unchanged.

[0096] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 12 mg of liraglutide, 8 mg of spleen aminopeptide, 6 mg of EDTA-2Na, 1 mg of glucose, 0.8 mg of benzyl alcohol, 5 mg of pH regulator, and the balance is water for injection.

[0097] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0098] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 32°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0099] (2) Dissolution of the stabilizer-enhancer: first add EDTA-2Na to the injection solution A obtained in step (1), stir and mix using a titanium rod stirring rod, then add liraglutide and mix to obtain injection solution B;

[0100] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.2 to obtain injection solution C;

[0101] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, then add glucose, and let stand for 25 minutes to obtain D injection;

[0102] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane;

[0103] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 12 min to obtain vitamin B12 injection solution.

[0104] Comparative Example 6

[0105] This comparative example differs from Example 3 in that glucose is not added to the ingredients, and the other ingredients and steps remain unchanged.

[0106] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 12 mg of liraglutide, 8 mg of spleen aminopeptide, 6 mg of L-cysteine, 0.8 mg of benzyl alcohol, 5 mg of pH regulator, and the balance is water for injection.

[0107] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0108] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 32°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0109] (2) Dissolution of stabilizer-enhancer: First, add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix using a titanium rod stirring rod, then add liraglutide and mix to obtain injection solution B;

[0110] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.2 to obtain injection solution C;

[0111] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, and let stand for 25 minutes to obtain D injection;

[0112] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane in sequence;

[0113] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 12 min to obtain vitamin B12 injection solution.

[0114] Comparative Example 7

[0115] This comparative example differs from Example 3 in that glucose is replaced with sodium chloride, and the other ingredients and steps remain unchanged.

[0116] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 12 mg of liraglutide, 8 mg of spleen aminopeptide, 6 mg of L-cysteine, 1 mg of sodium chloride, 0.8 mg of benzyl alcohol, 5 mg of pH regulator, and the balance is water for injection.

[0117] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0118] (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 32°C, and stir and dissolve them using a titanium stirring rod to obtain injection solution A;

[0119] (2) Dissolution of stabilizer-enhancer: First, add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix using a titanium stirring rod, then add liraglutide and mix to obtain injection solution B;

[0120] (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.2 to obtain injection solution C;

[0121] (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix with a titanium rod stirring rod, then add sodium chloride, and let stand for 25 minutes to obtain D injection;

[0122] (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection, and then sterilized by filtration using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane;

[0123] (6) Filling: Fill the filtered and sterilized injection solution into borosilicate glass ampoule (brown) according to the specifications, seal it, and sterilize it at 120°C for 12 min to obtain vitamin B12 injection solution.

[0124] Comparative Example 8

[0125] The difference between this comparative example and Example 3 is that the preparation method is to add the ingredients all at once without any order, and the other ingredients and steps remain unchanged.

[0126] A vitamin B12 injection, each mL of injection contains the following ingredients: 1.0 mg of vitamin B12, 12 mg of liraglutide, 8 mg of spleen aminopeptide, 6 mg of L-cysteine, 1 mg of glucose, 0.8 mg of benzyl alcohol, 5 mg of pH regulator, and the balance is water for injection.

[0127] A method for preparing a vitamin B12 injection, carried out under light-proof conditions, comprises the following steps:

[0128] Dissolution of raw materials: Weigh the prescribed amount of vitamin B12, benzyl alcohol, L-cysteine, liraglutide, spleen aminopeptide and glucose, add to water for injection at a temperature of 32°C, stir and dissolve with a titanium stirring rod, add a pH adjuster, adjust the pH value to 4.2, make up to volume with water for injection, and filter and sterilize using a 0.45μm polyethersulfone filter membrane and a 0.22μm polyethersulfone filter membrane in sequence; fill the filtered and sterilized injection into a borosilicate glass ampoule (brown) according to the specifications, seal and sterilize at a sterilization temperature of 120°C for 12 min to obtain vitamin B12 injection.

[0129] Comparative Example 9

[0130] Vitamin B12 injection is prepared according to the steps of the embodiments in the patent application No. 201310541073.7, entitled Vitamin B12 injection and its production process:

[0131] The filling process of vitamin B12 injection is carried out aseptically under class 100 laminar flow, and the sterilization step is eliminated. The drug solution is sterilized and filtered through a 0.22μm microporous filter membrane, and filled according to the filling job operation procedures. The sealing quality is checked at any time. The filling volume, visible foreign matter and residual oxygen content are randomly checked every 60 minutes. The filling volume is controlled at 1.00ml / vial, and the residual oxygen content is ≤2.0%.

[0132] Experimental Example 1 Composition Changes

[0133] 1.1 Select the vitamin B12 injection prepared in Examples 1 to 9, and use the reference preparation (ingredients: vitamin B12, sodium chloride, water for injection, manufacturer: Shandong Lukang Pharmaceutical Group Saite Co., Ltd., specification: 1 ml: 0.25 mg) as the control group to test the properties, impurities, content and stability of the vitamin B12 injection.

[0134] 1.2 Test Results

[0135] The results are shown in Table 1 and Table 2:

[0136] Table 1 Properties, impurities and content changes

[0137]

[0138] Table 2 Stability investigation

[0139]

[0140]

[0141] As can be seen from Table 1 and Table 2, the vitamin B12 injections prepared in Examples 1 to 3 all meet the requirements. In Table 1, Example 3 is compared with Comparative Examples 1 to 3 to show that spleen aminopeptide added to the injection as an impurity-reducing agent is beneficial to reducing the total impurity content in the injection, and liraglutide added to the injection as a synergist is beneficial to the stability of the effective content of vitamin B12; Example 3 is compared with Comparative Examples 4 to 7 to show that the present application has a certain selectivity for the ingredients in the vitamin B12 injection, and only by selecting the appropriate type of ingredients can the best effect be obtained. If a certain ingredient is missing or replaced with a similar ingredient, the experimental effect of Example 3 cannot be achieved; Example 3 is compared with Comparative Example 8 to show that the order of adding the ingredients in the preparation method of the vitamin B12 injection of the present application has a sequence. If the ingredients are not added in the prescribed order, the final injection product will have an increase in impurities and a low content of effective ingredients; at the same time, the injection prepared by Example 3 has the highest content of effective ingredients, indicating that the drug takes effect quickly after being injected into the body. In Table 2, the vitamin B12 injection prepared in Example 3 has higher stability than the reference preparation, and its total impurities increase slowly after long-term storage, and the content of active ingredients decreases slowly, which is conducive to long-term storage.

[0142] Experimental Example 2 Single Factor Investigation - Injection Water Temperature

[0143] 2.1 The temperature of the injection water during the production process will affect the dissolution rate of vitamin B12, which will cause the vitamin B12 raw materials to degrade during the preparation process, resulting in a high impurity content in the prepared vitamin B12 injection.

[0144] 2.2 Using the ingredient ratio of Example 3, vitamin B12 was dissolved in water for injection at different temperatures. Sample 1 was water for injection at 20°C, sample 2 was water for injection at 25°C, sample 3 was water for injection at 30°C, sample 4 was water for injection at 32°C, sample 5 was water for injection at 35°C, sample 6 was water for injection at 40°C, sample 7 was water for injection at 45°C, and sample 8 was water for injection at 50°C.

[0145] 2.3 Test results

[0146] As shown in Table 3:

[0147] Table 3 Detection of different injection water temperatures

[0148]

[0149]

[0150] As shown in Table 3, the total impurities are significantly reduced when the injection water temperature is between 30 and 35°C, especially at 32°C when the total impurity content is the lowest. Too high or too low injection water temperature can cause an increase in total impurities.

[0151] Experimental Example 3 Single Factor Investigation - Agitator Material

[0152] 3.1 During the production process, the liquid will directly contact materials such as stainless steel during the preparation, filtration, and filling processes, which may introduce impurities and affect the quality of the liquid. Therefore, the effect of using stirring rods of different materials during the preparation process on this product was investigated.

[0153] 3.2 Using the ingredient ratio of Example 3 and changing the material of the stirring rod, samples 9 to 12 were prepared, wherein sample 9 was a wooden stirring rod, sample 10 was a glass stirring rod, sample 11 was a silicone stirring rod, sample 12 was a stainless steel stirring rod, and the stirring rod of Example 3 was a titanium rod.

[0154] 3.3 Test results

[0155] As shown in Table 4:

[0156] Table 4 Agitator material test

[0157]

[0158] As shown in Table 4, wooden stirring rods, glass stirring rods and silicone stirring rods have no effect on the color of vitamin B12 injection, but they will introduce impurities and increase the total impurity content, which is not conducive to production and preparation; stainless steel stirring rods deepen the color of vitamin B12 injection and increase the total impurity content. Therefore, titanium stirring rods are required in the production process.

[0159] Experimental Example 4 Yield Calculation

[0160] The product yield of the vitamin B12 injection prepared in Example 3 and Comparative Examples 1 to 9 was calculated. The yield calculation formula is: yield = actual yield / theoretical yield. The results are shown in Table 5 as follows:

[0161] Table 5 Product yield

[0162]

[0163] As can be seen from the results in Table 5, since the vitamin B12 injection prepared in Comparative Examples 1 to 9 produces many bubbles in the liquid medicine during the filling process, it is easy to adhere to the ampoule bottle mouth when in contact with the ampoule bottle mouth, resulting in the carbonization of the liquid medicine adhering to the bottle mouth when the ampoule bottle is sealed at high temperature, making the injection solution unqualified with visible foreign matter, thereby reducing the product yield; and in Example 3, since the liquid medicine does not bubble and there is no situation of adhering to the ampoule bottle mouth, its yield has been greatly improved, especially in Comparative Examples 1, 3, 4 and 6, due to the lack of liraglutide, spleen aminopeptide, L-cysteine ​​and glucose, the composition changes, resulting in an unsatisfactory state of the liquid medicine, with a little adhesion at the bottle mouth, indicating that the use of complete ingredients is very important for the product yield.

[0164] Experimental Example 5 Safety Test of Injection

[0165] 5.1 The vitamin B12 injection prepared in Example 3 was used to perform an irritation test on the vitamin B12 injection of the present invention in accordance with the technical guidelines for drug irritation, allergy and hemolysis research in China. The experimental animals were rabbits, and a test group and a blank group (normal saline) were set. The test group and the blank group of this test were both subcutaneously injected at a dosing volume of 1.5 mg / kg on the back (rabbit hair removed), with a dose of 1.2 mg / kg, and 3 rabbits in each group.

[0166] 5.2 The injection site was observed with the naked eye before administration and 24h and 48h after administration, and it was found that the vitamin B12 injection of the present invention was non-irritating when used for subcutaneous injection and was safe to use.

[0167] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A vitamin B12 injection, characterized in that: Each mL of injection contains the following ingredients: vitamin B12 0.8-1.2 mg, liraglutide 10-15 mg, spleen aminopeptide 6-12 mg, L-cysteine ​​3-7 mg, glucose 0.5-5 mg, benzyl alcohol 0.3-1 mg, pH adjuster 1-10 mg, and the balance is water for injection.

2. The vitamin B12 injection according to claim 1, characterized in that Each mL of injection contains the following ingredients: vitamin B12 1.0 mg, liraglutide 12 mg, spleen aminopeptide 8 mg, L-cysteine ​​6 mg, glucose 1 mg, benzyl alcohol 0.8 mg, pH adjuster 5 mg, and the balance is water for injection.

3. The method for preparing vitamin B12 injection according to claim 1, characterized in that: The following steps are involved: (1) Dissolution of API: Weigh the prescribed amount of vitamin B12 and benzyl alcohol, add them into water for injection at a temperature of 30-35°C, stir and dissolve, and obtain injection solution A; (2) Dissolving the stabilizer-enhancer: first add L-cysteine ​​to the injection solution A obtained in step (1), stir and mix, then add liraglutide, mix and mix to obtain injection solution B; (3) pH adjustment: adding a pH adjuster to the injection solution B obtained in step (2) to adjust the pH value to 4.0-4.5 to obtain injection solution C; (4) Dissolution of the impurity reducing agent - osmotic pressure regulator: first add spleen aminopeptide to the C injection obtained in step (3), stir and mix, then add glucose, and let stand for 20 to 30 minutes to obtain D injection; (5) Volume adjustment and filtration sterilization: The D injection solution was adjusted to volume with water for injection and filtered for sterilization; (6) Filling: Fill, seal and sterilize the filtered and sterilized injection solution according to the specifications to obtain the vitamin B12 injection solution.

4. The method for preparing the vitamin B12 injection according to claim 3, characterized in that: The preparation method is carried out under light-proof conditions.

5. The method for preparing the vitamin B12 injection according to claim 3, characterized in that: The temperature of the water for injection in step (1) is 32°C.

6. The method for preparing the vitamin B12 injection according to claim 3, characterized in that: The stirring rod used for stirring is made of titanium.

7. The method for preparing the vitamin B12 injection according to claim 3, characterized in that: In step (3), the pH adjusting agent is 0.1 mol / L hydrochloric acid solution or 0.1 mol / L sodium hydroxide solution.

8. The method for preparing the vitamin B12 injection according to claim 3, characterized in that: In step (3), the pH value is adjusted to 4.

2.

9. The method for preparing the vitamin B12 injection according to claim 3, characterized in that: In step (5), filtration is performed using a 0.45 μm polyethersulfone filter membrane and a 0.22 μm polyethersulfone filter membrane in sequence.

10. The method for preparing vitamin B12 injection according to claim 3, characterized in that: The process parameters for sterilization in step (6) are a sterilization temperature of 120° C. and a sterilization time of 10 to 13 minutes.

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