Preparation method and application of compound amino acid injection

Abstract

The invention discloses a preparation method and application of a compound amino acid injection, and the preparation method comprises the following steps: adding water for injection into a closed container filled with protective gas; amino acids including tyrosine, leucine, isoleucine, phenylalanine, aspartic acid, glutamic acid, methionine, tryptophan, valine, threonine, histidine, alanine, glycine, arginine, lysine acetate, proline, serine and cysteine are sequentially added according to the following sequence, and liquid medicine is obtained; and heating and sterilizing the liquid medicine to obtain the compound amino acid injection. Specific amino acid components are added into the closed container filled with protective gas according to a specific sequence, a good anti-oxidation effect can be achieved under the condition that an antioxidant and an adsorbent are not added, and the 18-component compound amino acid injection which is high in active component content, low in impurity content, good in product quality stability and high in clinical use safety is obtained.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and in particular relates to a method for preparing a compound amino acid injection and its application. Background Technology

[0002] Compound Amino Acid Injection (18AA-III) is an amino acid-based intravenous nutrition drug used for clinical nutritional support, not limited to surgery. In the preparation of this type of product, if excessive sterilization processes are used without activated charcoal, problems such as low content of active ingredients, excessive degradation impurities, and yellowing of the solution during storage can easily occur, resulting in poor product quality stability. Amino acids are easily oxidized, and potential problems during preparation include: tryptophan is easily oxidized and degraded, leading to a darker solution color and an increase in related substances; methionine is easily oxidized and degraded to produce the impurity methionine sulfoxide; glutamic acid is easily affected by high temperatures, easily undergoing dehydration and condensation at high temperatures to form the degradation impurity pyroglutamic acid; and so on. In addition, this formulation usually contains the antioxidant sulfite, and the addition of antioxidants may cause allergic reactions such as asthma, resulting in poor clinical safety. Summary of the Invention

[0003] In order to overcome at least one of the problems existing in the prior art, one of the objectives of the present invention is to provide a method for preparing compound amino acid injection, which does not require the use of antioxidants and adsorbents in the preparation process, and the product has good quality stability and high safety for clinical use.

[0004] The second objective of this invention is to provide an application of the above-mentioned preparation method.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows: The first aspect of the present invention provides a method for preparing a compound amino acid injection, comprising the following steps: adding amino acids in the following order to a sealed container filled with water for injection and filled with a protective gas: tyrosine, leucine, isoleucine, phenylalanine, aspartic acid, glutamic acid, methionine, tryptophan, valine, threonine, histidine, alanine, glycine, arginine, lysine acetate, proline, serine, and cysteine ​​to obtain a drug solution; and sterilizing the drug solution by heating to obtain the compound amino acid injection.

[0006] In some embodiments of the present invention, the preparation method further includes the following step: controlling the headspace oxygen concentration of the sealed container to be ≤2% during the preparation process; for example, it can be any value or a range between any two of 0.1%, 0.3%, 0.5%, 0.7%, 1%, 1.3%, 1.5%, 1.7% or 2%.

[0007] In some embodiments of the present invention, the preparation method further includes the following step: controlling the dissolved oxygen concentration of the sealed container during the preparation process to be ≤2ppm; for example, it can be any value or a range between 0.1ppm, 0.3ppm, 0.5ppm, 0.7ppm, 1ppm, 1.3ppm, 1.5ppm, 1.7ppm or 2ppm.

[0008] The headspace oxygen concentration or dissolved oxygen concentration mentioned above can be adjusted by controlling the injection pressure and flow rate of the protective gas.

[0009] In some embodiments of the present invention, the protective gas includes at least one of nitrogen, argon, or helium; in some specific embodiments of the present invention, the protective gas is selected from nitrogen.

[0010] In some embodiments of the present invention, the preparation method further includes the following step: controlling the pH of the drug solution to be 5.2~6.8; for example, it can be any value or a range between any two of 5.2, 5.5, 5.7, 6, 6.3, 6.5 or 6.8; in some specific embodiments of the present invention, the preparation method further includes the following step: controlling the pH of the drug solution to be 5.5~6.5; in some more specific embodiments of the present invention, the preparation method further includes the following step: controlling the pH of the drug solution to be 5.7~6.3.

[0011] In some embodiments of the present invention, the reagents used to control the pH of the drug solution include acidic reagents, alkaline reagents, or combinations thereof; specifically, the acidic reagent may include at least one of hydrochloric acid, sulfuric acid, or acetic acid; the alkaline reagent may include at least one of ammonia, sodium hydroxide, or potassium hydroxide.

[0012] In some embodiments of the present invention, the heat sterilization temperature is 110~130℃; for example, it can be any value or a range between any two of 110℃, 113℃, 115℃, 117℃, 120℃, 121℃, 123℃, 125℃, 128℃, or 130℃; in some specific embodiments of the present invention, the heat sterilization temperature is 115~125℃; in some more specific embodiments of the present invention, the heat sterilization temperature is 117~123℃.

[0013] In some embodiments of the present invention, the heat sterilization time is 8 to 20 minutes; for example, it can be any value or a range between any two of 8 minutes, 9 minutes, 10 minutes, 11 minutes, 12 minutes, 13 minutes, 14 minutes, 15 minutes, 16 minutes, 18 minutes, or 20 minutes; in some specific embodiments of the present invention, the heat sterilization time is 10 to 14 minutes; in some more specific embodiments of the present invention, the heat sterilization time is 11 to 13 minutes.

[0014] In some embodiments of the present invention, the preparation method further includes the following step: before adding amino acids, controlling the temperature of the water for injection to 60~90℃; for example, it can be any value or a range between any two of 60℃, 62℃, 65℃, 67℃, 70℃, 72℃, 75℃, 80℃, 85℃, or 90℃; in some specific embodiments of the present invention, the preparation method further includes the following step: before adding amino acids, controlling the temperature of the water for injection to 62~80℃; in some more specific embodiments of the present invention, the preparation method further includes the following step: before adding amino acids, controlling the temperature of the water for injection to 65~75℃.

[0015] In some embodiments of the present invention, the preparation method further includes the following steps: before adding amino acids, the water for injection in the sealed container is 70-90% of the total mass of all water for injection; for example, it can be any value of 70%, 75%, 80%, 85% or 90% or a range between any two; after adding amino acids, the remaining water for injection is added.

[0016] In some embodiments of the present invention, the preparation method further includes the following step: filtering the drug solution before heating sterilization.

[0017] In some embodiments of the present invention, the preparation method further includes the following steps: filling the drug solution before heating sterilization; more specifically, the filling is done after nitrogen filling.

[0018] In some embodiments of the present invention, the medicinal solution comprises amino acids in the following amounts: Isoleucine 5.04~6.16 g / L; Leucine 11.25~13.75g / L; Lysine acetate 11.16~13.64 g / L; Methionine 3.15~3.85g / L; Phenylalanine: 8.41~10.29 g / L; Threonine 5.85~7.15 g / L; Tryptophan 1.17~1.43 g / L; Valine 4.05~4.95g / L; Alanine 5.58~6.82 g / L; Arginine 7.11~8.69 g / L; Aspartic acid 3.42~4.18 g / L; Glutamic acid 5.85~7.15 g / L; Histidine 5.40~6.60 g / L; Proline 2.97~3.63 g / L; Serine 1.98~2.42 g / L; Tyrosine 0.31~0.39 g / L; Glycine 9.63~11.77 g / L; Cysteine ​​0.90~1.10g / L.

[0019] In some specific embodiments of the present invention, the medicinal solution includes amino acids in the following amounts: Isoleucine 5.32~5.88 g / L; Leucine 11.88~13.13 g / L; Lysine acetate 11.78~13.02 g / L; Methionine 3.33~3.68 g / L; Phenylalanine: 8.88~9.82 g / L; Threonine 6.18~6.83 g / L; Tryptophan 1.24~1.37 g / L; Valine 4.28~4.73 g / L; Alanine 5.89~6.51 g / L; Arginine 7.51~8.30 g / L; Aspartic acid 3.61~3.99 g / L; Glutamic acid 6.18~6.83 g / L; Histidine 5.70~6.30 g / L; Proline 3.14~3.47 g / L; Serine 2.09~2.31 g / L; Tyrosine 0.33~0.37 g / L; Glycine 10.17~11.24 g / L; Cysteine ​​0.95~1.05g / L.

[0020] In some embodiments of the present invention, the amino acid composition of the drug solution is the same as that of compound amino acid injection 18AA-III.

[0021] In some embodiments of the present invention, the compound amino acid injection solution does not contain antioxidants.

[0022] In some embodiments of the present invention, no adsorbent is added during the preparation of the compound amino acid injection; in some specific embodiments of the present invention, the adsorbent includes activated carbon, ion exchange resin or a combination thereof.

[0023] In some embodiments of the present invention, the methionine sulfoxide content in the compound amino acid injection is 0.005-0.1% of the labeled amount of methionine; for example, it can be any value or a range between any two of 0.005%, 0.007%, 0.009%, 0.01%, 0.015%, 0.03%, 0.05%, 0.08%, or 0.1%; in some specific embodiments of the present invention, the methionine sulfoxide content in the compound amino acid injection is 0.007-0.05% of the labeled amount of methionine; in some more specific embodiments of the present invention, the methionine sulfoxide content in the compound amino acid injection is 0.009-0.015% of the labeled amount of methionine.

[0024] In some embodiments of the present invention, the pyroglutamic acid content in the compound amino acid injection is 1-5% of the labeled amount of glutamic acid; for example, it can be any value or a range between any two of 1%, 1.2%, 1.3%, 1.5%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, or 5%; in some specific embodiments of the present invention, the pyroglutamic acid content in the compound amino acid injection is 1.2-4% of the labeled amount of glutamic acid; in some more specific embodiments of the present invention, the pyroglutamic acid content in the compound amino acid injection is 1.3-3.5% of the labeled amount of glutamic acid.

[0025] In some embodiments of the present invention, the tyrosine content in the compound amino acid injection is 99-106% of the labeled amount; for example, it can be any value or a range between any two of 99%, 100%, 101%, 102%, 103%, 104%, 105%, or 106%; in some specific embodiments of the present invention, the tyrosine content in the compound amino acid injection is 100-105% of the labeled amount; in some more specific embodiments of the present invention, the tyrosine content in the compound amino acid injection is 102-104% of the labeled amount.

[0026] In some embodiments of the present invention, the tryptophan content in the compound amino acid injection is 99-106% of the labeled amount; for example, it can be any value or a range between any two of 99%, 100%, 101%, 102%, 103%, 104%, 105%, or 106%; in some specific embodiments of the present invention, the tryptophan content in the compound amino acid injection is 100-105% of the labeled amount; in some more specific embodiments of the present invention, the tryptophan content in the compound amino acid injection is 101-104% of the labeled amount.

[0027] In some embodiments of the present invention, the proline content in the compound amino acid injection is 99-106% of the labeled amount; for example, it can be any value or a range between any two of 99%, 100%, 101%, 102%, 103%, 104%, 105%, or 106%; in some specific embodiments of the present invention, the proline content in the compound amino acid injection is 101-105% of the labeled amount; in some more specific embodiments of the present invention, the proline content in the compound amino acid injection is 102-104% of the labeled amount.

[0028] In some embodiments of the present invention, the methionine content in the compound amino acid injection is 98-103% of the labeled amount; for example, it can be any value or a range between any two of 98%, 99%, 100%, 101%, 102%, or 103%; in some specific embodiments of the present invention, the methionine content in the compound amino acid injection is 99-102% of the labeled amount; in some more specific embodiments of the present invention, the methionine content in the compound amino acid injection is 100-101% of the labeled amount.

[0029] In some embodiments of the present invention, the glutamic acid content in the compound amino acid injection is 93-103% of the labeled amount; for example, it can be any value or a range between any two of 93%, 95%, 97%, 98%, 99%, 100%, 101%, 102%, or 103%; in some specific embodiments of the present invention, the glutamic acid content in the compound amino acid injection is 97-102% of the labeled amount; in some more specific embodiments of the present invention, the glutamic acid content in the compound amino acid injection is 99-101% of the labeled amount.

[0030] In some embodiments of the present invention, the cysteine ​​content in the compound amino acid injection is 89-100% of the labeled amount; for example, it can be any value or a range between any two of 89%, 90%, 92%, 94%, 95%, 96%, 98%, or 100%; in some specific embodiments of the present invention, the cysteine ​​content in the compound amino acid injection is 92-98% of the labeled amount; in some more specific embodiments of the present invention, the cysteine ​​content in the compound amino acid injection is 94-96% of the labeled amount.

[0031] A second aspect of the present invention provides the application of the preparation method as described in the first aspect of the present invention in the preparation of amino acid-based intravenous nutrition drugs.

[0032] The beneficial effects of this invention are: by adding specific amino acid components in a specific order in a sealed container equipped with a protective gas, this invention can achieve good antioxidant effects without adding antioxidants and adsorbents, resulting in an 18-component compound amino acid injection with high active ingredients, low impurity content, good product quality stability, and high safety for clinical use. Furthermore, the production process of this invention is simple and easy to control, and it has good application prospects in the preparation of amino acid-based intravenous nutrition drugs. Detailed Implementation

[0033] The following specific embodiments further illustrate the content of the present invention in detail. It should also be understood that the following embodiments are only for further explanation of the present invention and should not be construed as limiting the scope of protection of the present invention. Non-essential improvements and adjustments made by those skilled in the art based on the principles described herein are all within the scope of protection of the present invention. The specific process parameters, etc., in the following examples are merely examples within a suitable range; that is, those skilled in the art can make selections within a suitable range based on the description herein, and are not intended to be limited to the specific data in the examples below. Unless otherwise specified, the raw materials, reagents, or apparatus used in the following embodiments and comparative examples can be obtained from conventional commercial sources or by existing known methods.

[0034] The components and contents contained in each 1000 mL of the drug solution in Examples 1-4 and Comparative Example 2 are as follows: Ingredient content Isoleucine 5.60g; Leucine 12.50g; Lysine acetate 12.40g; Methionine 3.50g; Phenylalanine 9.35g; Threonine 6.50g; Tryptophan 1.30g; Valine 4.50g; Alanine 6.20g; Arginine 7.90g; Aspartic acid 3.80g; 6.50g of glutamic acid; Histidine 6.00g; Proline 3.30g; Serine 2.20g; Tyrosine 0.35g; Glycine 10.70g; Cysteine ​​1.00g; Add water for injection to a final volume of 1000 mL.

[0035] The difference between the drug solution in Comparative Example 1 and those in Examples 1-4 and Comparative Example 2 is that each 1000 mL of the drug solution also contains: Sodium bisulfite 0.50g; All other ingredients are the same.

[0036] Example 1 A method for preparing a compound amino acid injection, the specific steps of which are as follows: 1) Fill the sealed preparation tank and pipeline with nitrogen to control the headspace oxygen to below 2.0% and dissolved oxygen to below 2ppm. Add 80% of the prescribed amount of water for injection to the preparation tank and start feeding. 2) Control the temperature of the water for injection to 70℃, start stirring, and add tyrosine, leucine, isoleucine, phenylalanine, aspartic acid, glutamic acid, methionine, tryptophan, valine, threonine, histidine, alanine, glycine, arginine, lysine acetate, proline, serine, and cysteine ​​in sequence, stirring until dissolved; add water to the total volume, stir evenly, and obtain the drug solution; 3) The finished product is obtained by filtration, nitrogen filling, and sterilization at 121℃ / 12min.

[0037] Example 2 A method for preparing a compound amino acid injection, the specific steps of which are as follows: 1) Fill the sealed preparation tank and pipeline with nitrogen to control the headspace oxygen to below 2.0% and dissolved oxygen to below 2ppm. Add 80% of the prescribed amount of water for injection to the preparation tank and start feeding. 2) Control the temperature of the water for injection to 70℃, start stirring, and add tyrosine, leucine, isoleucine, phenylalanine, aspartic acid, glutamic acid, methionine, tryptophan, valine, threonine, histidine, alanine, glycine, arginine, lysine acetate, proline, serine, and cysteine ​​in sequence. Stir until dissolved, and adjust the pH to 6.8 with sodium hydroxide. Add water to the total volume, stir evenly, and obtain the drug solution. 3) The finished product is obtained by filtration, nitrogen filling, and sterilization at 121℃ / 12min.

[0038] Example 3 A method for preparing a compound amino acid injection, the specific steps of which are as follows: 1) Fill the sealed preparation tank and pipeline with nitrogen to control the headspace oxygen to below 2.0% and dissolved oxygen to below 2ppm. Add 80% of the prescribed amount of water for injection to the preparation tank and start feeding. 2) Control the temperature of the water for injection to 70℃, start stirring, and add tyrosine, leucine, isoleucine, phenylalanine, aspartic acid, glutamic acid, methionine, tryptophan, valine, threonine, histidine, alanine, glycine, arginine, lysine acetate, proline, serine, and cysteine ​​in sequence. Stir until dissolved, and adjust the pH to 5.2 with hydrochloric acid. Add water to the total volume, stir evenly, and obtain the drug solution. 3) The finished product is obtained by filtration, nitrogen filling, and sterilization at 121℃ / 12min.

[0039] Example 4 A method for preparing a compound amino acid injection, the specific steps of which are as follows: 1) Fill the sealed preparation tank and pipeline with nitrogen to control the headspace oxygen to below 2.0% and dissolved oxygen to below 2ppm. Add 80% of the prescribed amount of water for injection to the preparation tank and start feeding. 2) Control the temperature of the water for injection to 70℃, start stirring, and add tyrosine, leucine, isoleucine, phenylalanine, aspartic acid, glutamic acid, methionine, tryptophan, valine, threonine, histidine, alanine, glycine, arginine, lysine acetate, proline, serine, and cysteine ​​in sequence, stirring until dissolved; add water to the total volume, stir evenly, and obtain the drug solution; 3) The finished product is obtained by filtration, nitrogen filling, and sterilization at 121℃ for 15 minutes.

[0040] Comparative Example 1 A method for preparing a compound amino acid injection, the specific steps of which are as follows: 1) Add 80% of the prescribed amount of water for injection to a sealed preparation vessel and begin feeding: 2) Control the temperature of the water for injection to 70℃, start stirring, and add sodium bisulfite, tyrosine, leucine, isoleucine, phenylalanine, aspartic acid, glutamic acid, methionine, tryptophan, valine, threonine, histidine, alanine, glycine, arginine, lysine acetate, proline, serine, and cysteine ​​in sequence, stirring until dissolved; add water to the total volume, stir evenly, and obtain the drug solution; 3) Add needle-grade activated carbon and stir; filter to remove carbon; after fine filtration, nitrogen filling, and sterilization at 100℃ / 30min, the finished product is obtained.

[0041] Comparative Example 2 A method for preparing a compound amino acid injection, the specific steps of which are as follows: 1) Fill the sealed preparation tank and pipeline with nitrogen to control the headspace oxygen to below 2.0% and dissolved oxygen to below 2ppm. Add 80% of the prescribed amount of water for injection to the preparation tank and start feeding. 2) Control the temperature of the water for injection to 70℃, start stirring, and add tyrosine, leucine, aspartic acid, glutamic acid, methionine, isoleucine, phenylalanine, tryptophan, histidine, alanine, glycine, arginine, valine, threonine, lysine acetate, proline, serine, and cysteine ​​in sequence, stirring until dissolved; add water to the total volume, stir evenly, and obtain the drug solution; 3) The finished product is obtained by filtration, nitrogen filling, and sterilization at 121℃ / 12min.

[0042] Performance testing Detection methods: The contents of related substances, methionine sulfoxide, pyroglutamic acid, tyrosine and tryptophan were determined by high performance liquid chromatography (Chinese Pharmacopoeia 2025 Edition, Part IV, General Chapter 0512); the contents of proline, methionine, glutamic acid and cysteine ​​were determined by an amino acid analyzer.

[0043] The test results are shown in Table 1. The standards for related substances, methionine sulfoxide, and pyroglutamic acid are also shown in Table 1.

[0044] Table 1. Test results of each embodiment and comparative example.

[0045] As shown in Table 1, the finished products prepared in Examples 1-4 have low contents of related substances and low contents of degradation impurities, while having high contents of amino acid active ingredients. Among them, compared with other examples, Example 1 has the best overall effect, with a total content of related substances and impurities as low as 0.06%, methionine sulfoxide content as low as 0.01%, pyroglutamic acid content as low as 3.0%, and tyrosine content as high as 102.8%, tryptophan content as high as 103.1%, proline content as high as 103.0%, methionine content as high as 100.7%, glutamic acid content as high as 99.3%, and cysteine ​​content as high as 95.0%.

[0046] Comparative Example 1 added the antioxidant sodium bisulfite and the adsorbent activated carbon. Although the content of related substances and degradation impurities was effectively reduced, the activated carbon easily adsorbed the raw materials, resulting in a significant reduction in the content of amino acid active ingredients. Its product quality stability was poor, and the antioxidant it contained could easily trigger allergic problems such as asthma, making its clinical use safety poor.

[0047] Comparative Example 2 used a different order of amino acid addition than Example 1. The content of degradation impurity methionine sulfoxide in its finished product was significantly increased, and the content of tyrosine was significantly decreased. The content of other amino acids also decreased. The antioxidant effect was obviously not as good as that of Example 1, and the product quality stability was poor.

[0048] In summary, this invention, by adding specific amino acid components in a specific order within a sealed container equipped with a protective gas, can achieve excellent antioxidant effects without the addition of antioxidants and adsorbents, resulting in an 18-component compound amino acid injection with high active ingredients, low impurity content, good product quality stability, and high clinical safety. Furthermore, the production process of this invention is simple and easy to control, and it has good application prospects in the preparation of amino acid-based intravenous nutrition drugs.

Claims

1. A method for preparing a compound amino acid injection, characterized by, The method comprises the following steps: The method comprises the following steps:

2. The production method according to claim 1, characterized by, The method further comprises the following steps: controlling the headspace oxygen concentration of the closed container during the preparation process to be less than or equal to 2%; And / or, controlling the dissolved oxygen concentration of the closed container during the preparation process to be less than or equal to 2 ppm.

3. The preparation method according to claim 1, characterized in that, The method further comprises the following step: controlling the pH of the liquid to be 5.2-6.

8.

4. The method of claim 1, wherein, The temperature of the heat sterilization is 110-130°C; And / or, the heat sterilization time is 8-20 min.

5. The preparation method according to claim 1, characterized in that, The method further comprises the following step: before adding the amino acids, controlling the temperature of the water for injection to be 60-90°C.

6. The method of claim 1, wherein, The method further comprises the following steps: before adding the amino acids, the water for injection in the closed container accounts for 70-90% of the total mass of the water for injection; and after adding the amino acids, the remaining water for injection is added.

7. The preparation method according to claim 1, characterized in that, The liquid comprises the following amounts of amino acids: Isoleucine 5.04-6.16 g / L; Leucine 11.25-13.75 g / L; Lysine acetate 11.16-13.64 g / L; Methionine 3.15-3.85 g / L; Phenylalanine 8.41-10.29 g / L; Threonine 5.85-7.15 g / L; Tryptophan 1.17-1.43 g / L; Valine 4.05-4.95 g / L; Alanine 5.58-6.82 g / L; Arginine 7.11-8.69 g / L; Aspartic acid 3.42-4.18 g / L; Glutamic acid 5.85-7.15 g / L; Histidine 5.40-6.60 g / L; Proline 2.97-3.63 g / L; Serine 1.98-2.42 g / L; Tyrosine 0.31-0.39 g / L; Glycine 9.63-11.77 g / L; Cysteine 0.90-1.10 g / L.

8. The method of claim 1, wherein, The compound amino acid injection does not contain an antioxidant; And / or, no adsorbent is added during the preparation process of the compound amino acid injection.

9. The method of claim 1, wherein, The methionine sulfoxide content in the compound amino acid injection is 0.005-0.1% of the methionine label amount; And / or, the pyroglutamic acid content in the compound amino acid injection is 1-5% of the glutamic acid label amount; And / or, the tyrosine content in the compound amino acid injection is 99-106% of the label amount; And / or, the tryptophan content in the compound amino acid injection is 99-106% of the label amount; And / or, the proline content in the compound amino acid injection is 99-106% of the label amount; And / or, the methionine content in the compound amino acid injection is 98-103% of the label amount; And / or, the glutamic acid content in the compound amino acid injection is 93-103% of the labeled amount; And / or, the cysteine content in the compound amino acid injection is 89-100% of the labeled amount.

10. Use of the preparation method according to any one of claims 1-9 in the preparation of amino acid intravenous nutritional drugs.

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