Compound amino acid injection, stabilizer and preparation method thereof

Abstract

The application provides a stabilizer for a compound amino acid injection, a compound amino acid injection containing the stabilizer and a preparation method of the compound amino acid injection. The stabilizer is composed of cysteine and cystine in a weight ratio of (5-80):1. The compound amino acid injection contains the stabilizer and an amino acid component. The preparation method of the compound amino acid injection comprises the following steps: preparing a first amino acid solution under the condition of nitrogen protection; adjusting the pH value of the first amino acid solution to neutral, adding cysteine and cystine, dissolving the remaining water for injection to obtain a second amino acid solution; sterilizing the second amino acid solution to obtain the compound amino acid injection. The application uses cysteine and cystine as the stabilizer of the compound amino acid injection, so that the impurities of other amino acids are stabilized, and the safety is high.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparation technology, specifically relating to a stabilizer for compound amino acid injection, a compound amino acid injection containing the stabilizer, and a method for preparing the compound amino acid injection. Background Technology

[0002] Compound amino acid injections are a type of parenteral nutrition drug that plays an important role in nutritional therapy. They are widely used for patients who cannot receive oral nutrition or whose nutritional needs cannot be met through oral administration. To ensure the stability of the amino acids in compound amino acid injection products, sulfites, such as sodium bisulfite, are usually added as antioxidants.

[0003] However, sulfites pose potential toxic risks, specifically inducing hypersensitivity reactions and damaging tissues and organs. Therefore, long-term use of amino acid injections containing sulfites may cause rashes, itching, and in severe cases, even anaphylactic shock. Furthermore, it may cause damage to organs such as the liver and kidneys.

[0004] Currently, some studies have attempted to use other substances to replace sulfites to stabilize compound amino acid injections. For example, patent application CN102861011A discloses a pharmaceutical composition and formulation containing 18 amino acids, in which EDTA and cysteine ​​hydrochloride are selected as antioxidants for the compound amino acid injection. However, EDTA is an ion chelating agent that can bind to calcium ions, causing calcium to be chelated from bones, thereby leading to osteoporosis.

[0005] Therefore, providing a stabilizer and compound amino acid injection that are both highly safe and effective in stabilizing specific amino acid impurities is a pressing technical problem that needs to be solved. Summary of the Invention

[0006] To address the above problems, the purpose of this invention is to provide a stabilizer for compound amino acid injection, a compound amino acid injection containing the stabilizer, and a method for preparing the compound amino acid injection.

[0007] The above-mentioned objective of the present invention is achieved by providing the following technical solution:

[0008] In a first aspect, the present invention provides a stabilizer for compound amino acid injection, which is composed of cysteine ​​and cystine, wherein the weight ratio of cysteine ​​to cystine is (5-80):1.

[0009] According to some embodiments of the present invention, the weight ratio of cysteine ​​to cystine is (10-80):1, preferably (10-55):1, and more preferably (15-55):1.

[0010] Secondly, the present invention provides a compound amino acid injection containing a stabilizer and amino acid components other than cysteine ​​and cystine as described in the present invention.

[0011] According to some embodiments of the present invention, the weight ratio of the stabilizer to the amino acid components other than cysteine ​​and cystine is 1:(60-150); preferably, the weight ratio of the stabilizer to the amino acid components is 1:(70-130).

[0012] According to some embodiments of the present invention, the compound amino acid injection contains 0.81 to 1.44 g of the stabilizer in a total volume of 1000 ml; preferably, the compound amino acid injection contains 0.81 to 1.21 g of the stabilizer; more preferably, the compound amino acid injection contains 0.81 to 1.05 g of the stabilizer.

[0013] According to some embodiments of the present invention, the compound amino acid injection contains 0.80–1.20 g cysteine ​​and 0.01–0.24 g cysteine, based on a total volume of 1000 ml; preferably, the compound amino acid injection contains 0.80–1.10 g cysteine ​​and 0.01–0.11 g cysteine; more preferably, the compound amino acid injection contains 0.80–1.10 g cysteine ​​and 0.02–0.11 g cysteine; and even more preferably, the compound amino acid injection contains 0.80–1.00 g cysteine ​​and 0.02–0.05 g cysteine.

[0014] According to some embodiments of the present invention, the compound amino acid injection contains 80-125g of amino acid components other than cysteine ​​and cystine, based on a total volume of 1000ml; preferably, the compound amino acid injection contains 82-123g of amino acid components other than cysteine ​​and cystine; more preferably, the compound amino acid injection contains 90-115g of amino acid components other than cysteine ​​and cystine.

[0015] According to some embodiments of the present invention, the amino acid components other than cysteine ​​and cystine include isoleucine, leucine, lysine acetate, methionine, phenylalanine, threonine, tryptophan, valine, alanine, arginine, aspartic acid, glutamic acid, histidine, proline, serine, tyrosine, and glycine.

[0016] According to some embodiments of the present invention, the compound amino acid injection solution, in a total volume of 1000 ml, comprises 6.40–9.60 g isoleucine, 11.20–16.80 g leucine, 11.84–17.76 g lysine acetate, 3.12–4.68 g methionine, 5.60–8.40 g phenylalanine, 4.56–6.84 g threonine, 1.60–2.40 g tryptophan, and 6.40–9.60 g valine. 6.40–9.60g alanine, 6.72–10.08g arginine, 0.80–1.20g aspartic acid, 0.80–1.20g glutamic acid, 4.00–6.00g histidine, 4.00–6.00g proline, 2.40–3.60g serine, 0.40–0.60g tyrosine, 4.72–7.08g glycine, 0.80–1.20g cysteine, and 0.01–0.24g cystine.

[0017] Preferably, based on a total volume of 1000 ml, the compound amino acid injection contains 7.20–8.80 g isoleucine, 12.60–15.40 g leucine, 13.32–16.28 g lysine acetate, 3.51–4.29 g methionine, 6.30–7.70 g phenylalanine, 5.13–6.27 g threonine, 1.80–2.20 g tryptophan, 7.20–8.80 g valine, and 7.2 g amino acids. 0–8.80g alanine, 7.56–9.24g arginine, 0.90–1.10g aspartic acid, 0.90–1.10g glutamic acid, 4.50–5.40g histidine, 4.50–5.40g proline, 2.70–3.30g serine, 0.45–0.55g tyrosine, 5.31–6.49g glycine, 0.80–1.10g cysteine, and 0.01–0.11g cystine.

[0018] More preferably, based on a total volume of 1000 ml, the compound amino acid injection contains 7.20–8.80 g isoleucine, 12.60–15.40 g leucine, 13.32–16.28 g lysine acetate, 3.51–4.29 g methionine, 6.30–7.70 g phenylalanine, 5.13–6.27 g threonine, 1.80–2.20 g tryptophan, 7.20–8.80 g valine, and 7.2 g amino acids. 0–8.80g alanine, 7.56–9.24g arginine, 0.90–1.10g aspartic acid, 0.90–1.10g glutamic acid, 4.50–5.40g histidine, 4.50–5.40g proline, 2.70–3.30g serine, 0.45–0.55g tyrosine, 5.31–6.49g glycine, 0.80–1.10g cysteine, and 0.02–0.11g cysteine.

[0019] More preferably, based on a total volume of 1000 ml, the compound amino acid injection contains 8.00 g isoleucine, 14.00 g leucine, 14.80 g lysine acetate, 3.90 g methionine, 7.00 g phenylalanine, 5.70 g threonine, 2.00 g tryptophan, 8.00 g valine, 8.00 g alanine, 8.40 g arginine, 1.00 g aspartic acid, 1.00 g glutamic acid, 5.00 g histidine, 5.00 g proline, 3.00 g serine, 0.50 g tyrosine, 5.90 g glycine, 1.00 g cysteine, and 0.05 g cysteine.

[0020] According to some embodiments of the present invention, the pH value of the compound amino acid injection is 6.5 to 7.5, preferably 7.0.

[0021] Thirdly, the present invention provides a method for preparing the compound amino acid injection according to the present invention, which includes the following steps:

[0022] (1) Under nitrogen protection, add some water for injection to the amino acid components other than cysteine ​​and cystine, and dissolve at 60-80°C to obtain the first amino acid solution;

[0023] (2) Adjust the pH of the first amino acid solution to neutral, add cysteine ​​and cystine, dissolve them, and then add the remaining water for injection to obtain the second amino acid solution;

[0024] (3) Sterilize the second amino acid solution to obtain the compound amino acid injection;

[0025] Alternatively, it may include the following steps:

[0026] (1) Under nitrogen protection, add some water for injection to the amino acid components other than glutamic acid, cysteine ​​and cystine, and dissolve at 60-80°C to obtain the first amino acid solution;

[0027] (2) The temperature of the first amino acid solution was lowered to below 40°C and its pH value was adjusted to neutral. Glutamic acid, cysteine ​​and cystine were added, dissolved and the remaining water for injection was added to obtain the second amino acid solution.

[0028] (3) Sterilize the second amino acid solution to obtain compound amino acid injection.

[0029] According to some embodiments of the present invention, in step (1), the portion of water for injection is 70% to 90% of the total water for injection by weight.

[0030] According to some embodiments of the present invention, in step (1), the nitrogen protection condition is to seal and continuously fill with nitrogen to maintain a positive nitrogen pressure.

[0031] According to some embodiments of the present invention, in step (1), the dissolved oxygen of the first amino acid solution is controlled to be <300 ppb by continuous stirring and nitrogen purging.

[0032] According to some embodiments of the present invention, in step (2), glacial acetic acid is used to adjust the pH of the first amino acid solution to 6.5 to 7.5.

[0033] According to some embodiments of the present invention, in step (2), the dissolved oxygen in the second amino acid solution is <300 ppb.

[0034] According to some embodiments of the present invention, in steps (1) and (2), the water for injection is water for injection that has undergone deoxygenation treatment.

[0035] According to some embodiments of the present invention, in step (3), the second amino acid solution is sterilized after filling; preferably, the second amino acid solution is filtered before filling; more preferably, a polyethersulfone filter is used for filtration; more preferably, the pore size of the polyethersulfone filter is 0.22µm.

[0036] According to some embodiments of the present invention, in step (3), the headspace oxygen of the filled second amino acid injection is controlled to be less than 0.5%.

[0037] According to some embodiments of the present invention, in step (3), the sterilization is carried out by hot water spray sterilization; preferably, the sterilization conditions are a sterilization temperature of 121±2℃, a sterilization time of 8 to 12 min, and an F0 value greater than 8.

[0038] Fourthly, the present invention provides the use of the compound amino acid injection according to the present invention in the preparation of a medicament for amino acid supplementation in patients with hypoproteinemia, low nutritional status and / or pre- and post-operative conditions.

[0039] Fifthly, the present invention provides the use of the stabilizer according to the present invention in the preparation of a medicament for improving the stability of a compound amino acid preparation.

[0040] Compared with existing technologies, the technical solution of the present invention has the following beneficial effects:

[0041] 1. This invention uses a combination of cysteine ​​and cystine as a stabilizer in compound amino acid injections to stabilize other amino acids. Since both cysteine ​​and cysteine ​​are natural amino acids, they are relatively safe as stabilizers and have a good effect on reducing specific impurities.

[0042] 2. Experimental verification shows that, compared to the combination of cysteine ​​and sodium bisulfite, the stabilizer of this invention achieves comparable or even better stabilization effects, while significantly improving safety. Compared to other stabilizers, such as the combination of cysteine ​​and EDTA, the stabilizer of this invention significantly improves the performance of compound amino acid injections in reducing several specific impurities.

[0043] 3. Experimental results show that the compound amino acid injection of the present invention has good clarity and transmittance. Detailed Implementation

[0044] The present invention will be further described in detail below with reference to specific embodiments. The embodiments given are only for illustrating the present invention and are not intended to limit the scope of the present invention.

[0045] Unless otherwise specified, the experimental methods used in the following examples and comparative examples are conventional methods. Unless otherwise specified, the reagents used in the following examples and comparative examples are commercially available products.

[0046]

[0047] Example 1

[0048] The prescription for the compound amino acid injection in this embodiment is shown in Table 1.

[0049]

[0050] The preparation process of the compound amino acid injection in this embodiment is as follows:

[0051] Weigh the prescribed amounts of amino acid raw materials according to Table 1 and set aside. Under positive nitrogen pressure, add the amino acid raw materials (16 amino acids excluding glutamic acid, cysteine, and cystine), seal and continuously purge with nitrogen. Add deoxygenated water for injection to the preparation tank to 90% of the prepared volume. Maintain the temperature at 65-70°C, start stirring, and continuously purge with nitrogen, controlling dissolved oxygen <300 ppb. Cool to 40°C, add glacial acetic acid to adjust the pH (6.5-7.5 (target value at 25°C: 7.0)), and add glutamic acid, cysteine, and cystine. Rinse the tank walls with deoxygenated water at 40°C and stir to dissolve. Control dissolved oxygen (less than 300 ppb), and add deoxygenated water for injection to the preparation tank to full volume. Filter using a polyethersulfone filter (0.22µm pore size). After filling the solution, evacuate each bottle several times with nitrogen, then cap and seal, controlling headspace oxygen to below 0.5%. Sterilization was performed by hot water spraying under the following conditions: sterilization temperature (121℃), sterilization time (12 min), and F0 value (12). The product was then packaged after light inspection.

[0052] Example 2

[0053] The prescription for the compound amino acid injection in this embodiment is shown in Table 2.

[0054]

[0055] The preparation process of the compound amino acid injection in this embodiment is as follows:

[0056] Weigh the prescribed amounts of amino acid raw materials according to Table 2 and set aside. Under positive nitrogen pressure, add the amino acid raw materials (17 amino acids excluding cysteine ​​and cystine), seal and continuously purge with nitrogen. Add deoxygenated water for injection to the preparation tank to 70% of the prepared volume. Maintain the temperature at 60-65℃, start stirring, and continuously purge with nitrogen, controlling dissolved oxygen <300ppb. Add glacial acetic acid to adjust the pH (6.5-7.5 (target value at 25℃: 7.0)), and add cysteine ​​and cystine. Rinse the tank walls with deoxygenated water and stir to dissolve. Control dissolved oxygen (less than 300ppb), and add deoxygenated water for injection to the preparation tank to full volume. Filter using a polyethersulfone filter (0.22µm pore size). After filling the solution, evacuate each bottle several times with nitrogen, then cap and seal, controlling headspace oxygen to below 0.5%. Sterilization was performed by hot water spraying under the following conditions: sterilization temperature (121℃), sterilization time (12 min), and F0 value (12). The product was then packaged after light inspection.

[0057] Example 3

[0058] The prescription for the compound amino acid injection in this embodiment is shown in Table 3.

[0059]

[0060] The preparation process of the compound amino acid injection in this embodiment is as follows:

[0061] Weigh the prescribed amounts of amino acid raw materials according to Table 3 and set aside. Under positive nitrogen pressure, add the amino acid raw materials (16 amino acids excluding glutamic acid, cysteine, and cystine), seal and continuously purge with nitrogen. Add deoxygenated water for injection to the preparation tank to 80% of the prepared volume. Maintain the temperature at 70-75°C, start stirring, and continuously purge with nitrogen, controlling dissolved oxygen <300 ppb. Add glacial acetic acid to adjust the pH (6.5-7.5 (target value at 25°C: 7.0)), cool to 30°C, and then add glutamic acid, cysteine, and cystine sequentially. Rinse the tank walls with deoxygenated water and stir to dissolve. Control dissolved oxygen (less than 300 ppb), and add deoxygenated water for injection to the preparation tank to full volume. Filter using a polyethersulfone filter (0.22µm pore size). After filling the solution, evacuate each vial several times with nitrogen, then cap and seal, controlling headspace oxygen to below 0.5%. Sterilization was performed by hot water spraying under the following conditions: sterilization temperature (121℃), sterilization time (8 min), and F0 value (8). The product was then packaged after light inspection.

[0062] Examples 4 to 5

[0063] The formulations of the compound amino acid injections in Examples 4 and 5 are shown in Table 4.

[0064]

[0065] The preparation process of the compound amino acid injection in Examples 4 and 5 is basically the same as that in Example 1, except that the dosage of each amino acid is adjusted according to the prescription in Table 4.

[0066] Comparative Example 1

[0067] The compound amino acid injection in this comparative example uses cysteine ​​and sodium bisulfite as antioxidants and stabilizers, and the prescription is shown in Table 5.

[0068]

[0069] The preparation process of the compound amino acid injection in this comparative example is as follows:

[0070] Weigh the prescribed amounts of amino acid raw materials according to Table 5 and set aside. Under nitrogen positive pressure protection, add the 16 amino acid raw materials (excluding glutamic acid and cysteine) to the preparation tank, seal and continuously purge with nitrogen, and add 70% of the prepared amount of deoxygenated water for injection. Maintain the temperature at 65 to 70°C, start stirring, and continuously purge with nitrogen, controlling dissolved oxygen <300 ppb. Stir until completely dissolved. Cool to 40°C, add glacial acetic acid to adjust the pH (6.5~7.5 (target value at 25°C: 7.0)), add sodium bisulfite, glutamic acid, and cysteine, rinse the tank wall with a small amount of deoxygenated water, and stir to dissolve. Control dissolved oxygen (less than 300 ppb), and add deoxygenated water for injection to the preparation tank until full volume. Filter using a polyethersulfone filter (0.22µm pore size). After filling the drug solution, evacuate each bottle several times with nitrogen, then cap and seal, controlling headspace oxygen below 0.5%. Sterilization was performed by hot water spraying under the following conditions: sterilization temperature (121℃), sterilization time (8 min), and F0 value (8). The product was then packaged after light inspection.

[0071] Comparative Example 2

[0072] The compound amino acid injection in this comparative example uses cysteine ​​+ EDTA as a stabilizer, and the prescription is shown in Table 6.

[0073]

[0074] The preparation process of the compound amino acid injection in this comparative example is as follows:

[0075] Weigh the prescribed amounts of amino acid raw materials according to Table 6 and set aside. Under positive nitrogen pressure, add the 17 amino acid raw materials (excluding cysteine) to the preparation tank, seal and continuously purge with nitrogen. Add deoxygenated water for injection to the preparation tank to 80% of the prepared volume. Maintain the temperature at 70-75℃, start stirring, continuously purge with nitrogen, control dissolved oxygen <300ppb, add glacial acetic acid and EDTA to adjust the pH (6.5-7.5 (target value at 25℃: 7.0)), and add cysteine. Rinse the tank wall with a small amount of deoxygenated water and stir to dissolve. Control dissolved oxygen (less than 300ppb), and add deoxygenated water for injection to the preparation tank to the full volume. Filter using a polyethersulfone filter (0.22µm pore size). After filling the drug solution, evacuate each bottle several times with nitrogen, then cap and seal, controlling headspace oxygen to below 0.5%. Sterilization was carried out by hot water spraying. The sterilization conditions were: sterilization temperature (121℃), sterilization time (12min), and F0 value (12). The product was packaged after light inspection.

[0076] Comparative Example 3

[0077] The compound amino acid injection in this comparative example only contains cysteine ​​as a stabilizer, and the prescription is shown in Table 7.

[0078]

[0079] The preparation process of the compound amino acid injection in this comparative example is as follows:

[0080] Weigh the prescribed amounts of amino acid raw materials according to Table 7 and set aside. Under positive nitrogen pressure, add the 16 amino acid raw materials (excluding glutamic acid and cysteine) to the preparation tank, seal and continuously purge with nitrogen. Add deoxygenated water for injection to the preparation tank to 80% of the prepared volume. Maintain the temperature at 70-75℃, start stirring, and continuously purge with nitrogen, controlling dissolved oxygen <300ppb. Stir until completely dissolved. Cool to 40℃, add glacial acetic acid to adjust the pH (6.5-7.5 (target value at 25℃: 7.0)), and add glutamic acid and cysteine. Rinse the tank wall with a small amount of deoxygenated water and stir to dissolve. Control dissolved oxygen (less than 300ppb), and add deoxygenated water for injection to the preparation tank to full volume. Take samples for monitoring. Filter using a polyethersulfone filter (0.22µm pore size). After filling the drug solution, evacuate each bottle several times with nitrogen, then cap and seal, controlling headspace oxygen to below 0.5%. Sterilization was carried out by hot water spraying. The sterilization conditions were: sterilization temperature (121℃), sterilization time (12min), and F0 value (12). The product was packaged after light inspection.

[0081] Comparative Example 4

[0082] The compound amino acid injection in this comparative example has cysteine ​​and acetylcysteine ​​added as stabilizers, and the prescription is shown in Table 8.

[0083]

[0084] The preparation process of the compound amino acid injection in this comparative example is as follows:

[0085] Weigh the prescribed amounts of amino acid raw materials according to Table 8 and set aside. Under positive nitrogen pressure, add the amino acid raw materials (16 amino acids excluding glutamic acid, cysteine, and acetylcysteine), seal and continuously purge with nitrogen. Add deoxygenated water for injection to the preparation tank to 90% of the prepared volume. Maintain the temperature at 65-70°C, start stirring, and continuously purge with nitrogen, controlling dissolved oxygen <300 ppb. Cool to 40°C, add glacial acetic acid to adjust the pH (6.5-7.5 (target value at 25°C: 7.0)), and add glutamic acid, cysteine, and acetylcysteine. Rinse the tank walls with deoxygenated water at 40°C and stir to dissolve. Control dissolved oxygen (less than 300 ppb), and add deoxygenated water for injection to the preparation tank to full volume. Filter using a polyethersulfone filter (0.22µm pore size). After filling the solution, evacuate each bottle several times with nitrogen, then cap and seal, controlling headspace oxygen to below 0.5%. Sterilization was performed by hot water spraying under the following conditions: sterilization temperature (121℃), sterilization time (12 min), and F0 value (12). The product was then packaged after light inspection.

[0086] Comparative Example 5

[0087] The compound amino acid injection in this comparative example has cysteine ​​and cystine added as stabilizers, and the prescription is shown in Table 9.

[0088]

[0089] The preparation process of the compound amino acid injection in this comparative example is as follows:

[0090] Weigh the prescribed amounts of amino acid raw materials according to Table 9 and set aside. Under positive nitrogen pressure, add the amino acid raw materials (16 amino acids excluding glutamic acid, cysteine, and cystine), seal and continuously purge with nitrogen. Add deoxygenated water for injection to the preparation tank to 80% of the prepared volume. Maintain the temperature at 70-75°C, start stirring, and continuously purge with nitrogen, controlling dissolved oxygen <300 ppb. Add glacial acetic acid to adjust the pH (6.5-7.5 (target value at 25°C: 7.0)), cool to 30°C, and then add glutamic acid, cysteine, and cystine sequentially. Rinse the tank walls with deoxygenated water and stir to dissolve. Control dissolved oxygen (less than 300 ppb), and add deoxygenated water for injection to the preparation tank to full volume. Filter using a polyethersulfone filter (0.22µm pore size). After filling the solution, evacuate each bottle several times with nitrogen, then cap and seal, controlling headspace oxygen to below 0.5%. Sterilization was performed by hot water spraying under the following conditions: sterilization temperature (121℃), sterilization time (8 min), and F0 value (8). The product was then packaged after light inspection.

[0091] Stability test

[0092] 1. Investigation of influencing factors: Samples were taken and tested after being placed at 50℃ for 10 and 30 days;

[0093] Accelerated testing conditions: Samples were taken and tested after being stored at 40℃ for 1 month and 3 months.

[0094] 2. Methionine sulfoxide and pyroglutamic acid

[0095] Analytical method: Determined according to high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0096] Test solution: Accurately measure 2 ml of this product and place it in a 10 ml volumetric flask. Dilute with water to the mark and shake well.

[0097] Reference solution: Take appropriate amounts of methionine sulfoxide and pyroglutamic acid reference standards, accurately weigh them, dissolve them in water, and quantitatively dilute them to prepare a solution containing 4 μg of methionine sulfoxide and 8 μg of pyroglutamic acid per 1 ml.

[0098] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase. Mobile phase A was phosphate buffer (prepared as an aqueous solution of 0.03 mol / L potassium dihydrogen phosphate and 0.01 mol / L sodium octanesulfonate, adjusted to pH 1.5 with phosphoric acid solution) – acetonitrile (95:5 v / v), and mobile phase B was acetonitrile. Gradient elution was performed according to the table below. The flow rate was 1.0 mL / min; the column temperature was 35 °C; and the detection wavelength was 205 nm.

[0099]

[0100] Assay: Accurately measure the test solution and the reference solution, inject them separately into the liquid chromatograph, and record the chromatograms.

[0101] 3. Tyrosine, phenylalanine, methionine, and tryptophan

[0102] Analytical method: Determined according to high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0103] Test solution: Accurately measure 1 ml of this product, place it in a 20 ml volumetric flask, dilute with water to the mark, and shake well.

[0104] Reference solution: Weigh appropriate amounts of tyrosine, phenylalanine, methionine and tryptophan reference standards accurately, dissolve in water and dilute quantitatively to prepare a mixed solution containing 25 μg of tyrosine, 350 μg of phenylalanine, 195 μg of methionine and 100 μg of tryptophan per ml.

[0105] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase; 0.03 mol / L triethylamine aqueous solution (pH adjusted to 2.0 with phosphoric acid) - methanol (volume ratio 95:5) was used as the mobile phase; the flow rate was 1.0 mL per minute; the column temperature was 30 °C; and the detection wavelength was 280 nm.

[0106] System suitability requirements: The resolution of each peak in the chromatogram of the reference solution should meet the requirements, and the theoretical plate number based on the tryptophan peak should not be less than 2000.

[0107] Assay: Accurately measure the test solution and the reference solution, inject them separately into the liquid chromatograph, and record the chromatograms.

[0108] 4. Dioxindole alanine, kynurenine, 2-hydroxytryptophan

[0109] Analytical method: Determined according to high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0110] Test solution: Accurately measure 2 ml of this product and place it in a 10 ml volumetric flask. Dilute with water to the mark and shake well.

[0111] Reference solution: Weigh appropriate amounts of dioxindole alanine, kynurenine, and 2-hydroxytryptophan reference standards accurately, dissolve in water, and quantitatively dilute to prepare a mixed solution containing 0.8 μg of dioxindole alanine, 0.8 μg of kynurenine, and 0.8 μg of 2-hydroxytryptophan per 1 ml.

[0112] Chromatographic conditions: Octadecyl-bonded silica gel was used as the stationary phase; phosphate buffer (prepared as 0.02 mol / L diammonium hydrogen phosphate solution, pH adjusted to 8.0 with ammonia)-methanol (95:5 v / v) was used as mobile phase A, and water-acetonitrile (50:50 v / v) was used as mobile phase B, with gradient elution according to the table below; flow rate was 1.0 mL / min; column temperature was 30 °C; detection wavelength was 210 nm.

[0113]

[0114] Assay: Accurately measure the test solution and the reference solution, inject them separately into the liquid chromatograph, and record the chromatograms.

[0115] 5. Phenylethylamine

[0116] Analytical method: Determined according to high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0117] Test solution: Accurately measure 3 ml of this product, place it in a 20 ml volumetric flask, dilute with water to the mark, and shake well.

[0118] Reference solution: Weigh an appropriate amount of phenethylamine reference standard accurately, dissolve it in water and dilute quantitatively to prepare a solution containing 1 μg of phenethylamine per 1 ml.

[0119] System suitability solution: Take 3 ml of this product and an appropriate amount of phenethylamine reference standard, place them in 20 ml, and dilute with water to prepare a mixed solution containing this product and 1 μg / ml phenethylamine.

[0120] Chromatographic conditions: Octadecyl bonded silica gel was used as the stationary phase; phosphate buffer (prepared as an aqueous solution of 0.03 mol / L dipotassium hydrogen phosphate and 0.01 mol / L sodium octane sulfonate, with pH adjusted to 4.0 by phosphoric acid) - methanol (volume ratio 70:30) was used as the mobile phase; the flow rate was 0.5 mL / min; the column temperature was 30 °C; and the detection wavelength was 210 nm.

[0121] Assay: Accurately measure the test solution and the reference solution, inject them separately into the liquid chromatograph, and record the chromatograms.

[0122] 6. Clarity and color of compound amino acid injection: Take this product and determine according to the method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0901, Method I and General Chapter 0902, Method I). The solution should be clear and colorless. If color is developed, it should not be darker than the yellow No. 2 standard colorimetric solution.

[0123] 7. Transmittance of Compound Amino Acid Injection: Take this product and determine the transmittance according to the ultraviolet-visible spectrophotometry method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0401) at a wavelength of 430 nm. The transmittance shall not be less than 97.0%.

[0124] The experimental results of Examples 1 to 5 and Comparative Examples 1 to 5 are shown in Tables 10 to 19, respectively.

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133]

[0134]

[0135] The above results indicate that when cysteine ​​+ sodium bisulfite is used as an antioxidant, tryptophan impurities are significantly increased and exceed limits, while other impurities remain at low levels, and the color of the amino acid injection changes slightly. When cysteine ​​+ EDTA is used as an antioxidant, some tryptophan impurities are significantly increased and exceed limits, pyroglutamic acid content is slightly increased, and the color of the amino acid injection changes slightly. When cysteine ​​is used alone as an antioxidant, tryptophan and methionine impurities are significantly increased and exceed limits, pyroglutamic acid content is significantly increased and exceeds limits, and the color of the amino acid injection changes significantly, becoming a yellow liquid. When cysteine ​​+ acetylcysteine ​​is used as an antioxidant, tryptophan impurities are significantly increased and exceed limits, pyroglutamic acid content is significantly increased, and the color of the amino acid injection changes significantly, becoming a yellow liquid. When cysteine ​​and cysteine ​​are used as antioxidants, but the ratio of the two components is not within the scope of protection of this invention, tryptophan impurities are significantly increased and exceed limits, pyroglutamic acid content is significantly increased, and the color of the amino acid injection changes significantly, becoming a yellow liquid.

[0136] The compound amino acid injection solution using the stabilizer of this invention remained a colorless and transparent liquid under accelerated conditions, with stable pH and transmittance. The content of easily degradable amino acids such as methionine, tyrosine, phenylalanine, and tryptophan remained stable and all exceeded 98% by weight. After 3 months under accelerated conditions, the content of impurities such as methionine sulfoxide, dioxindole alanine, kynurenine, 2-hydroxytryptophan, phenylethylamine, and pyroglutamic acid all met the standard requirements, and the impurity content was significantly lower compared to comparative proportions using cysteine ​​+ sodium bisulfite, cysteine ​​+ EDTA, cysteine ​​+ acetylcysteine, and cysteine. These results demonstrate that the stabilizer of this invention can achieve better impurity reduction and improved stability.

[0137] The above descriptions are merely several exemplary embodiments of the present invention and are not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any equivalent or related embodiments obtained by those skilled in the art through some modifications or variations made to the above-disclosed technical content without departing from the scope of the present invention are within the scope of the present invention.

Claims

1. A compound amino acid injection, wherein, Based on a total volume of 1000 ml, the compound amino acid injection contains 6.40–9.60 g isoleucine, 11.20–16.80 g leucine, 11.84–17.76 g lysine acetate, 3.12–4.68 g methionine, 5.60–8.40 g phenylalanine, 4.56–6.84 g threonine, 1.60–2.40 g tryptophan, 6.40–9.60 g valine, and 6.40–9.60 g lysine. 0.60g alanine, 6.72-10.08g arginine, 0.80-1.20g aspartic acid, 0.80-1.20g glutamic acid, 4.00-6.00g histidine, 4.00-6.00g proline, 2.40-3.60g serine, 0.40-0.60g tyrosine, 4.72-7.08g glycine, 0.80-1.20g cysteine, and 0.01-0.24g cysteine.

2. The compound amino acid injection according to claim 1, wherein, Based on a total volume of 1000 ml, the compound amino acid injection contains 7.20–8.80 g isoleucine, 12.60–15.40 g leucine, 13.32–16.28 g lysine acetate, 3.51–4.29 g methionine, 6.30–7.70 g phenylalanine, 5.13–6.27 g threonine, 1.80–2.20 g tryptophan, 7.20–8.80 g valine, and 7.20– 8.80g alanine, 7.56–9.24g arginine, 0.90–1.10g aspartic acid, 0.90–1.10g glutamic acid, 4.50–5.40g histidine, 4.50–5.40g proline, 2.70–3.30g serine, 0.45–0.55g tyrosine, 5.31–6.49g glycine, 0.80–1.10g cysteine, and 0.01–0.11g cystine.

3. The compound amino acid injection according to claim 1, wherein, Based on a total volume of 1000 ml, the compound amino acid injection contains 7.20–8.80 g isoleucine, 12.60–15.40 g leucine, 13.32–16.28 g lysine acetate, 3.51–4.29 g methionine, 6.30–7.70 g phenylalanine, 5.13–6.27 g threonine, 1.80–2.20 g tryptophan, 7.20–8.80 g valine, and 7.20– 8.80g alanine, 7.56–9.24g arginine, 0.90–1.10g aspartic acid, 0.90–1.10g glutamic acid, 4.50–5.40g histidine, 4.50–5.40g proline, 2.70–3.30g serine, 0.45–0.55g tyrosine, 5.31–6.49g glycine, 0.80–1.10g cysteine, and 0.02–0.11g cystine.

4. The compound amino acid injection according to claim 1, wherein, Based on a total volume of 1000 ml, the compound amino acid injection contains 8.00 g isoleucine, 14.00 g leucine, 14.80 g lysine acetate, 3.90 g methionine, 7.00 g phenylalanine, 5.70 g threonine, 2.00 g tryptophan, 8.00 g valine, 8.00 g alanine, 8.40 g arginine, 1.00 g aspartic acid, 1.00 g glutamic acid, 5.00 g histidine, 5.00 g proline, 3.00 g serine, 0.50 g tyrosine, 5.90 g glycine, 1.00 g cysteine, and 0.05 g cysteine.

5. The compound amino acid injection according to any one of claims 1 to 4, wherein, The pH value of the compound amino acid injection is 6.5 to 7.

5.

6. The compound amino acid injection according to claim 5, wherein, The pH value of the compound amino acid injection is 7.

0.

7. The method for preparing the compound amino acid injection according to any one of claims 1 to 6, comprising the following steps: (1) Under nitrogen protection, add some water for injection to the amino acid components other than cysteine ​​and cystine, and dissolve at 60-80°C to obtain the first amino acid solution; (2) Adjust the pH of the first amino acid solution to neutral, add cysteine ​​and cystine, dissolve them, and then add the remaining water for injection to obtain the second amino acid solution; (3) Sterilize the second amino acid solution to obtain compound amino acid injection.

8. A method for preparing the compound amino acid injection according to any one of claims 1 to 6, comprising the following steps: (1) Under nitrogen protection, add some water for injection to the amino acid components other than glutamic acid, cysteine ​​and cystine, and dissolve at 60-80°C to obtain the first amino acid solution; (2) The temperature of the first amino acid solution was lowered to below 40°C and its pH value was adjusted to neutral. Glutamic acid, cysteine ​​and cystine were added, dissolved and the remaining water for injection was added to obtain the second amino acid solution. (3) Sterilize the second amino acid solution to obtain compound amino acid injection.

9. The preparation method according to claim 7 or 8, wherein, In step (1), the portion of water for injection, by weight, is 70% to 90% of the total water for injection.

10. The preparation method according to claim 7 or 8, wherein, In step (1), the nitrogen protection condition is to seal and continuously fill with nitrogen to maintain a positive nitrogen pressure.

11. The preparation method according to claim 7 or 8, wherein, In step (1), the dissolved oxygen in the first amino acid solution is controlled to be <300 ppb by continuous stirring and nitrogen purging.

12. The preparation method according to claim 7 or 8, wherein, In step (2), glacial acetic acid is used to adjust the pH of the first amino acid solution to 6.5-7.

5.

13. The preparation method according to claim 7 or 8, wherein, In step (2), after adding cysteine ​​and cystine, the mixture is first rinsed with water for injection.

14. The preparation method according to claim 7 or 8, wherein, In step (2), the dissolved oxygen in the second amino acid solution is <300 ppb.

15. The preparation method according to claim 7 or 8, wherein, In steps (1) and (2), the water for injection is water for injection that has undergone deoxygenation treatment.

16. The preparation method according to claim 7 or 8, wherein, In step (3), the second amino acid solution is sterilized after being filled.

17. The preparation method according to claim 16, wherein the second amino acid solution is filtered before filling.

18. The preparation method according to claim 17, wherein, It uses a polyethersulfone filter cartridge for filtration.

19. The preparation method according to claim 18, wherein, The polyethersulfone filter element has a pore size of 0.22µm.

20. The preparation method according to claim 16, wherein, In step (3), the headspace oxygen of the filled second amino acid solution is controlled to be below 0.5%.

21. The preparation method according to claim 7 or 8, wherein, In step (3), the sterilization is carried out by hot water spray sterilization.

22. The preparation method according to claim 7 or 8, wherein, The sterilization conditions are a sterilization temperature of 121±2℃, a sterilization time of 8 to 12 minutes, and an F0 value greater than 8.

23. Use of the compound amino acid injection according to any one of claims 1 to 6 in the preparation of a medicament for amino acid supplementation in patients with hypoproteinemia, low nutritional status and / or pre- and post-operative conditions.

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