Compound amino acid injection and its application and preparation method

Abstract

A compound amino acid injection and its application and preparation method, the compound amino acid injection includes an amino acid composition and does not include a sulfite antioxidant, the amino acid composition includes a certain proportion of N (2) L-alanyl L-glutamine, acetylcysteine ​​and other active substances, each active substance exists in the form of a prototype, a pharmaceutically acceptable salt or a short peptide, and the ratio of each active substance is the same as the specification. The compound amino acid injection of the embodiment of the present application can obtain better stability without adding a sulfite antioxidant, can avoid sulfite antioxidants from causing harm to the human body, is suitable for treating or improving amino acid metabolism disorders and hypoproteinemia caused by trauma, burns, various surgical operations, severe infections or intestinal flora displacement, and improves the body's immune function and intestinal function.

Description

Technical Field

[0001] The present application relates to but is not limited to the field of medical technology, and in particular to a compound amino acid injection and its application and preparation method. Background Art

[0002] Amino acids are substrates for the body to synthesize proteins and other biologically active substances. Among them, lysine, threonine, leucine, isoleucine, valine, methionine, tryptophan, and phenylalanine are eight amino acids that cannot be synthesized by the human body and must be supplemented from outside the body. The basic requirement of amino acids for healthy adults is 0.8 to 1 g / (kg / d), but the amount of supplementation needs to be increased in cases of severe catabolism, obvious protein loss, or severe malnutrition.

[0003] For patients who cannot take amino acids orally or through the intestines, or whose amino acids cannot be met by oral or intestinal supplementation, intravenous infusion of compound amino acid injection is required to meet the body's needs for protein synthesis.

[0004] There are related reports on the research of amino acid solutions. For example, Chinese invention patent CN100518815C discloses an amino acid composition, and the amino acid injection disclosed in Example 1 includes 20 kinds of amino acids or dipeptides including acetylcysteine, and also contains an auxiliary component of sodium bisulfite. The efficacy test shows that the amino acid composition of Example 1 has good nutritional effects.

[0005] Chinese invention patent application CN1189167C discloses that amino acids are very unstable in solution and are easily oxidized to produce a series of complex substances that are harmful to the human body. Since the compound amino acids were put into production in the 1960s, sulfite compounds with unique antioxidant properties, such as sodium bisulfite and sodium pyrosulfite, must be added to ensure the stability of the product. Using sulfites (such as sodium bisulfite or sodium pyrosulfite, etc.) as antioxidants can protect amino acids from oxidative decomposition, which plays a key role in the stability of the product.

[0006] The 2020 edition of the Pharmacopoeia of the People's Republic of China contains six types of "Compound Amino Acid Injections". Although the amino acid formulas of these six "Compound Amino Acids" are different, each variety of prescription contains sulfite (sodium bisulfite or sodium metabisulfite).

[0007] Some unstable amino acids in amino acid infusion products, such as tryptophan, methionine, glutamic acid, acetylcysteine, etc., will degrade under the action of high temperature, oxygen, metal ions, etc., causing the solution to turn yellow, the amino acid content to decrease, and the related substances to increase, which increases the risk of clinical application of amino acid infusion products. Compound amino acid injection is a large-volume injection product that must be terminally sterilized. In order to ensure the stability of the product, many Chinese companies currently add sulfite as an antioxidant to their products.

[0008] However, sulfites are harmful to the human body. Many adverse reactions have been reported clinically. The most common one is sulfite allergy (especially in asthmatic patients), with symptoms such as bronchospasm, wheezing, dyspnea, malignant laryngeal edema, hypotension, shock and even death. In addition, some studies have reported that sulfites may damage chromosomes and DNA, and cause irreversible reactions of disulfide bonds in proteins.

[0009] Although the Chinese Pharmacopoeia clearly stipulates that if sodium bisulfite is used in injections, the general concentration is 0.1% to 0.2%, the risks of sodium bisulfite in clinical applications are still inevitable. Summary of the invention

[0010] The following is an overview of the subject matter described in detail herein. This overview is not intended to limit the scope of protection of this application.

[0011] In a first aspect, an embodiment of the present application provides a compound amino acid injection, wherein the compound amino acid injection comprises an amino acid composition and does not include a sulfite antioxidant, wherein the amino acid composition comprises the following active substances in a weight ratio, each active substance being present in the form of a prototype, a pharmaceutically acceptable salt or a short peptide:

[0012]

[0013]

[0014] In an embodiment of the present application, the amino acid composition may include the following active substances in the following weight ratios, each active substance being present in the form of original form, a pharmaceutically acceptable salt or a short peptide:

[0015] Aspartic acid 3.2-4.8 Isoleucine 6.0-9.0 Threonine 3.5-5.2 Leucine 10.4-15.6 Serine 4.0-6.0 Phenylalanine 3.0-4.5 Glutamate 1.3-1.9 Lysine acetate 5.6-8.5 Proline 4.0-6.0 Histidine 2.1-3.1 Glycine 2.6-3.8 Arginine 6.9-10.3 Alanine 0.5-0.7 Acetylcysteine 0.81-1.3 N(2)-L-Alanyl-L-Glutamine 14.4-21.7 Acetyl Tyrosine 0.81-1.1 Valine 7.6-11.4 Tryptophan 0.9-1.3 Methionine 2.6-3.8 Taurine 0.8-1.2

[0016] In an embodiment of the present application, the amino acid composition may include the following active substances in the following weight ratios, each active substance being present in the form of original form, a pharmaceutically acceptable salt or a short peptide:

[0017]

[0018]

[0019] In the embodiments of the present application, the compound amino acid injection does not include an antioxidant.

[0020] In a second aspect, the embodiments of the present application also provide the use of the compound amino acid injection provided in the first aspect of the embodiments of the present application in nutritional therapy or nutritional support.

[0021] In the embodiments of the present application, the nutritional therapy or nutritional support may be used to treat or improve amino acid metabolism disorders and hypoproteinemia, and enhance the body's immune function and intestinal function.

[0022] In the embodiments of the present application, the amino acid metabolism disorder and hypoproteinemia may be caused by trauma, burns, various surgical operations, severe infection or intestinal flora translocation.

[0023] In a third aspect, the present embodiment also provides a method for preparing the compound amino acid injection provided in the first aspect of the present embodiment, comprising:

[0024] S10: Cleaning and sterilization of auxiliary items for weighing;

[0025] S20: Cleaning and sterilization of auxiliary supplies for filling;

[0026] S30: Weighing and deoxygenation of APIs; including:

[0027] (1) Weighing the API;

[0028] (2) Place all the weighed APIs into a sealed powder mixing tank;

[0029] (3) performing multiple vacuum evacuations and nitrogen fillings on the powder mixing tank to remove oxygen-containing gas entrained between the API particles;

[0030] S40: Deoxygenation of water for injection; including:

[0031] (1) After the water for injection is prepared, the dissolved oxygen in the water for injection is removed;

[0032] (2) The deoxygenated water for injection is kept ready for use or transported to a storage tank through a closed pipeline, and the storage tank is continuously pressurized with nitrogen;

[0033] S50: Preparation of liquid medicine; including:

[0034] (1) Deoxygenate the liquid dispensing tank;

[0035] (2) under nitrogen protection, the water for injection to be used in step (2) of S40 or the water for injection in the storage tank is transported through a closed pipeline to the liquid preparation tank maintained under nitrogen pressure;

[0036] (3) The API processed in step (3) of S30 and the water for injection in the liquid preparation tank are respectively discharged through the discharge port of the powder mixing tank and the discharge port of the liquid preparation tank in a certain proportion, and mixed and dissolved in a mixer, and the dissolved liquid is returned to the liquid preparation tank through the discharge port of the mixer; the liquid is circulated multiple times between the liquid preparation tank and the mixer, and the API in the powder mixing tank is continuously transported to the mixer during the multiple circulations of the liquid until the API in the mixing tank is completely transported to the mixer;

[0037] (4) Cooling and sterilizing the liquid medicine prepared in step (3) of S50, and transferring the cooled and filtered liquid medicine to a deoxygenated product tank under nitrogen protection, continuously passing nitrogen into the product tank containing the liquid medicine and controlling the temperature in the product tank;

[0038] S60: Filling; including:

[0039] (1) performing secondary sterilization filtration on the drug liquid in the product tank, and filling it into a drug container that has been depyrogenated and cooled under nitrogen protection, pre-filling the drug container with nitrogen before filling to expel oxygen in the drug container, and filling with nitrogen during filling to reduce oxygen intrusion;

[0040] (2) Vacuuming and nitrogen filling the medicine container with liquid medicine several times and then sealing it with a plug;

[0041] (3) rolling the cap on the sealed pharmaceutical container;

[0042] S70: terminal sterilization, lamp inspection, headspace residual oxygen detection and packaging to obtain the compound amino acid injection.

[0043] In the embodiments of the present application, the residual oxygen value of the powder mixing tank after vacuuming and nitrogen filling, the liquid preparation tank maintained by nitrogen pressure, and the product tank after deoxygenation can be ≤1000ppm.

[0044] In an embodiment of the present application, the dissolved oxygen content in the injection water obtained in step (1) of S40 may be less than 40 ppb.

[0045] In an embodiment of the present application, the temperature for dissolving the API in step (3) of S50 may be 55°C to 65°C.

[0046] In an embodiment of the present application, in step (4) of S50, the drug solution prepared in step (3) of S50 may be cooled to 35°C to 45°C.

[0047] In an embodiment of the present application, in step (4) of S50, the primary sterilization filtration may include filtering using filters with a diameter of 0.45 μm and a diameter of 0.2 μm respectively.

[0048] In an embodiment of the present application, in step (4) of S50, the temperature in the product tank may be controlled to be no higher than 50°C.

[0049] In an embodiment of the present application, in step (1) of S60, the secondary sterilization filtration may include filtering the drug solution in the product tank with nitrogen, and sterilizing and filtering the filtered drug solution using a polyethersulfone sterilization filter with a diameter of 0.2 μm.

[0050] In an embodiment of the present application, step (1) of S60 may further include depyrogenating and cooling the medicine bottle, wherein the depyrogenating temperature is 330°C to 340°C, and the cooling includes a first stage cooling at a temperature ≤50°C and a second stage cooling at a temperature ≤35°C.

[0051] In an embodiment of the present application, in step (2) of S60, the number of times of vacuuming and nitrogen filling may be no less than 5 times.

[0052] In an embodiment of the present application, in step S70, the terminal sterilization temperature requires an F0 value greater than 8.

[0053] In an embodiment of the present application, in step S70, the headspace residual oxygen value in the medicine container is required to be no higher than 1%.

[0054] Compared with the currently available compound amino acid injection (for example, the injection disclosed in Example 1 of Chinese invention patent CN100518815C), the compound amino acid injection provided in the embodiment of the present application can obtain better stability without adding any antioxidant. For example, acetylcysteine ​​has better stability in the amino acid solution, which can avoid the harm of sulfite antioxidants to the human body.

[0055] The preparation method of the compound amino acid injection in the embodiment of the present application does not use activated carbon, so it can obtain better light transmittance and avoid the product risks caused by the use of activated carbon.

[0056] Other features and advantages of the present application will be described in the following description, and partly become apparent from the description, or be understood by implementing the present application. Other advantages of the present application can be realized and obtained by the schemes described in the description and the drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] The accompanying drawings are used to provide an understanding of the technical solution of the present application and constitute a part of the specification. Together with the embodiments of the present application, they are used to explain the technical solution of the present application and do not constitute a limitation on the technical solution of the present application.

[0058] Figure 1 This is the curve of the change of glutamine concentration in the blood of patients undergoing laparotomy with the operation time, where group A is the study group and group B is the control group;

[0059] Figure 2 Figure 2 is the curve of the change of glutamine concentration in the blood of laparoscopic patients with operation time, in which series 1 is the study group and series 2 is the control group. DETAILED DESCRIPTION

[0060] In order to make the purpose, technical solution and advantages of the present application more clear, the embodiments of the present application will be described in detail below with reference to the accompanying drawings. It should be noted that the embodiments and features in the embodiments of the present application can be combined with each other arbitrarily without conflict.

[0061] In the description of the present application, the weight ratio or content of the active substance is given in the form in which the active substance exists in the compound amino acid injection, for example,

[0062] In the description of the present application, the weight ratio of the active substance or the content of the active substance is given based on the form in which the active substance exists in the compound amino acid injection (original form, pharmaceutically acceptable salt or short peptide). For example, when an active substance exists in the compound amino acid injection in the form of a pharmaceutically acceptable salt, the weight of the active substance given refers to the weight of its pharmaceutically acceptable salt in the compound amino acid injection.

[0063] In the description of the present application, the compound amino acid injection does not contain an antioxidant, which means that it does not contain an additional antioxidant. The additional antioxidant here does not include the active substances with antioxidant effects among the 20 active substances in the amino acid composition. For example, although acetylcysteine ​​has an antioxidant effect, it does not belong to an additional antioxidant.

[0064] In a first aspect, an embodiment of the present application provides a compound amino acid injection, the compound amino acid injection comprising an amino acid composition and excluding a sulfite antioxidant, the amino acid composition including but not limited to the following active substances in weight proportions, each active substance being present in the form of a prototype, a pharmaceutically acceptable salt or a short peptide:

[0065] Aspartic acid 1-5.0 Isoleucine 3-10 Threonine 2.5-5.4 Leucine 5-16 Serine 4.0-10 Phenylalanine 2.5-5 Glutamate 1-15 Lysine acetate 3.0-9.0 Proline 4.0-10 Histidine 1.5-3.5 Glycine 0.1-8 Arginine 6.5-15 Alanine 0-5 Acetylcysteine 0.81-2 N(2)-L-Alanyl-L-Glutamine 8-30 Acetyl Tyrosine 0.81-4 Valine 5-12 Tryptophan 0.9-1.6 Methionine 2.5-4 Taurine 0-3

[0066] In the embodiments of the present application, the amino acid composition may be composed of the following active substances in the following weight ratios, that is, the following table is a specific formula of the amino acid composition, and each active substance exists in the form of a prototype, a pharmaceutically acceptable salt or a short peptide:

[0067]

[0068]

[0069] In the embodiments of the present application, the amino acid composition may include, but is not limited to, the following weight ratios of active substances, each active substance being present in the form of original form, a pharmaceutically acceptable salt or a short peptide:

[0070] Aspartic acid 3.2-4.8 Isoleucine 6.0-9.0 Threonine 3.5-5.2 Leucine 10.4-15.6 Serine 4.0-6.0 Phenylalanine 3.0-4.5 Glutamate 1.3-1.9 Lysine acetate 5.6-8.5 Proline 4.0-6.0 Histidine 2.1-3.1 Glycine 2.6-3.8 Arginine 6.9-10.3 Alanine 0.5-0.7 Acetylcysteine 0.81-1.3 N(2)-L-Alanyl-L-Glutamine 14.4-21.7 Acetyl Tyrosine 0.81-1.1 Valine 7.6-11.4 Tryptophan 0.9-1.3 Methionine 2.6-3.8 Taurine 0.8-1.2

[0071] In the embodiments of the present application, the amino acid composition may be composed of the following active substances in the following weight ratios, that is, the following table is a specific formula of the amino acid composition, and each active substance exists in the form of a prototype, a pharmaceutically acceptable salt or a short peptide:

[0072]

[0073]

[0074] In the embodiments of the present application, the amino acid composition may include, but is not limited to, the following weight ratios of active substances, each active substance being present in the form of original form, a pharmaceutically acceptable salt or a short peptide:

[0075] Aspartic acid 4.00 Isoleucine 7.50 Threonine 4.32 Leucine 13.00 Serine 5.00 Phenylalanine 3.75 Glutamate 1.60 Lysine acetate 7.05 Proline 5.00 Histidine 2.60 Glycine 3.20 Arginine 8.60 Alanine 0.60 Acetylcysteine 1.21 N(2)-L-Alanyl-L-Glutamine 18.05 Acetyl Tyrosine 0.92 Valine 9.50 Tryptophan 1.08 Methionine 3.20 Taurine 1.00

[0076] In the embodiments of the present application, the amino acid composition may be composed of the following active substances in the following weight ratios, that is, the following table is a specific formula of the amino acid composition, and each active substance exists in the form of a prototype, a pharmaceutically acceptable salt or a short peptide:

[0077]

[0078]

[0079] Compared with the currently available compound amino acid injection (for example, the injection disclosed in Example 1 of Chinese invention patent CN100518815C), the compound amino acid injection provided in the embodiment of the present application can obtain better stability without adding any antioxidant, for example, the stability of acetylcysteine ​​in the amino acid solution is better.

[0080] In the embodiments of the present application, the content of the active substance in the amino acid composition in the compound amino acid injection may be:

[0081]

[0082] In the embodiments of the present application, the content of the active substance in the amino acid composition in the compound amino acid injection may be:

[0083]

[0084] In the embodiments of the present application, the content of the active substance in the amino acid composition in the compound amino acid injection may be:

[0085]

[0086]

[0087] In the description of the present application, each active substance in the amino acid composition may exist in the form of a prototype, a suitable pharmaceutically acceptable salt or a short peptide. For example, the active substance may exist in the form of a pharmaceutically acceptable salt such as hydrochloride, acetate, etc.; for another example, the active substance may exist in the form of a short peptide such as glycyl-L-glutamine, L-cystinyl-L-glutamine, L-tyrosyl-L-glutamine, L-leucyl-L-glutamine, etc.

[0088] In the amino acid composition of the compound amino acid injection provided in the first aspect of the embodiment of the present application, aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, phenylalanine, lysine acetate, histidine, arginine, tryptophan, taurine, acetylcysteine, alanylglutamine, etc. are already included in Part II of the Pharmacopoeia of the People's Republic of China (2020 edition). Acetylcysteine ​​is also called N-acetyl-L-cysteine, acetyltyrosine is also called N-acetyl-L-tyrosine, and alanylglutamine is also called L-alanyl-L-glutamine or N(2)-L-alanyl-L-glutamine.

[0089] In a second aspect, the embodiments of the present application also provide the use of the compound amino acid injection provided in the first aspect of the embodiments of the present application in nutritional therapy or nutritional support.

[0090] In the embodiments of the present application, the nutritional therapy or nutritional support is used to treat or improve amino acid metabolism disorders and hypoproteinemia, and to enhance the body's immune function and intestinal function.

[0091] In the embodiments of the present application, the amino acid metabolism disorder and hypoproteinemia are caused by trauma, burns, various surgical operations, severe infection or intestinal flora translocation.

[0092] In the embodiments of the present application, the compound amino acid injection can be administered parenterally.

[0093] In the embodiments of the present application, other auxiliary components may be added to the compound amino acid injection, such as EDTA, citric acid and other commonly used auxiliary materials acceptable to the human body, or sorbitol, xylitol, glucose and other sugar sources.

[0094] The compound amino acid injection of the embodiment of the present application can be used for nutritional therapy or nutritional support to improve the nutritional status of patients, for example, it can be used for trauma, burns, various surgical operations, severe infection, intestinal flora translocation and amino acid metabolism disorders and hypoproteinemia caused by various reasons, to improve the body's immune function and intestinal function.

[0095] The compound amino acid injection of the embodiment of the present application can be an injection for parenteral administration. The inner packaging of the injection in direct contact with the drug can be a suitable packaging such as a bottle that meets pharmaceutical requirements, a double-chamber bag, a multi-chamber bag packaging, or a multi-layer co-extruded infusion film (bag). When packaged with a double-chamber bag, a multi-chamber bag, or a multi-layer co-extruded infusion film (bag): the components of the amino acid composition of the compound amino acid injection of the embodiment of the present application can be placed in the same cavity (layer), or can be placed in different cavities (layers); the components of the amino acid composition of the compound amino acid injection of the embodiment of the present application can exist partially or completely in solid form, or can be partially or completely dissolved in a suitable solvent such as water.

[0096] In a third aspect, the present embodiment also provides a method for preparing the compound amino acid injection provided in the first aspect of the present embodiment, comprising:

[0097] S10: Cleaning and sterilization of auxiliary items for weighing;

[0098] S20: Cleaning and sterilization of auxiliary supplies for filling;

[0099] S30: Weighing and deoxygenation of APIs; including:

[0100] (1) Weighing the API;

[0101] (2) Place all the weighed APIs into a sealed powder mixing tank;

[0102] (3) performing multiple vacuum evacuations and nitrogen fillings on the powder mixing tank to remove oxygen-containing gas entrained between the API particles;

[0103] S40: Deoxygenation of water for injection; including:

[0104] (1) After the water for injection is prepared, the dissolved oxygen in the water for injection is removed;

[0105] (2) The deoxygenated water for injection is kept ready for use or transported to a storage tank through a closed pipeline, and the storage tank is continuously pressurized with nitrogen;

[0106] S50: Preparation of drug solution; including:

[0107] (1) Deoxygenate the liquid dispensing tank;

[0108] (2) under nitrogen protection, the water for injection to be used in step (2) of S40 or the water for injection in the storage tank is transported through a closed pipeline to the liquid preparation tank maintained under nitrogen pressure;

[0109] (3) The API processed in step (3) of S30 and the water for injection in the liquid preparation tank are respectively discharged through the discharge port of the powder mixing tank and the discharge port of the liquid preparation tank in a certain proportion, and mixed and dissolved in a mixer, and the dissolved liquid is returned to the liquid preparation tank through the discharge port of the mixer; the liquid is circulated multiple times between the liquid preparation tank and the mixer, and the API in the powder mixing tank is continuously transported to the mixer during the multiple circulations of the liquid until the API in the mixing tank is completely transported to the mixer;

[0110] (4) Cooling and sterilizing the liquid medicine prepared in step (3) of S50, and transferring the cooled and filtered liquid medicine to a deoxygenated product tank under nitrogen protection, continuously passing nitrogen into the product tank containing the liquid medicine and controlling the temperature in the product tank;

[0111] S60: Filling; including:

[0112] (1) performing secondary sterilization filtration on the drug liquid in the product tank, and filling it into a drug container that has been depyrogenated and cooled under nitrogen protection, pre-filling the drug container with nitrogen before filling to expel oxygen in the drug container, and filling with nitrogen during filling to reduce oxygen intrusion;

[0113] (2) Vacuuming and nitrogen filling the medicine container with liquid medicine several times and then sealing it with a plug;

[0114] (3) rolling the cap on the sealed pharmaceutical container;

[0115] S70: terminal sterilization, lamp inspection, headspace residual oxygen detection and packaging to obtain the compound amino acid injection.

[0116] Activated carbon is used as a decolorizing agent in the preparation process of almost all the compound amino acid injections that have been disclosed so far, in order to ensure the light transmittance of the product. However, since the quality control of activated carbon itself is difficult, its use in high-risk varieties such as injections increases product risks. To this end, the Drug Review Center of the China National Drug Administration issued the "Technical Requirements for the Quality and Efficacy Consistency Evaluation of Generic Chemical Injections" in 2020, which stipulates: "It is recommended not to use activated carbon in the production of injections." According to this technical requirement, the inventors of this application removed the activated carbon from the production process of the compound amino acid injection. Surprisingly, the compound amino acid injection made after removing the activated carbon has no difference in light transmittance compared with the compound amino acid injection without removing the activated carbon.

[0117] In the embodiments of the present application, the residual oxygen value of the powder mixing tank after vacuuming and nitrogen filling, the liquid preparation tank maintained by nitrogen pressure, and the product tank after deoxygenation can be ≤1000ppm.

[0118] In an embodiment of the present application, in step (1) of S40, a degassing tower may be used to remove dissolved oxygen in the injection water; and the temperature in the degassing tower may be no less than 121°C.

[0119] In an embodiment of the present application, the dissolved oxygen content in the injection water obtained in step (1) of S40 may be less than 40 ppb.

[0120] In an embodiment of the present application, the temperature for dissolving the raw material in step (3) of S50 can be 55°C to 65°C, for example, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 64°C or 65°C; the dissolution time can be within 2 hours, for example, 0.5 hour to 2 hours or 1 hour to 2 hours, and for example, 0.5 hour, 1 hour, 1.5 hours, 2 hours, etc.

[0121] In an embodiment of the present application, the mixer used in step (3) of S50 can be a three-way mixer. The discharge port of the liquid preparation tank and the discharge port of the powder mixing tank can be connected to the two feed ports of the three-way mixer, and the discharge port of the three-way mixer is connected to the feed port of the liquid preparation tank through a nitrogen-filled pipeline, thereby realizing the dissolution of the bulk drug and multiple circulations of the liquid obtained by the dissolution. Through multiple circulations, the mixing of multiple bulk drugs can be made more uniform, and the dissolution in water for injection can be more sufficient.

[0122] In an embodiment of the present application, in step (4) of S50, the medicinal solution prepared in step (3) of S50 can be cooled to 35°C to 45°C, for example, it can be cooled to 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, 41°C, 42°C, 43°C, 44°C or 45°C.

[0123] In an embodiment of the present application, in step (4) of S50, the primary sterilization filtration may include filtering using filters with a diameter of 0.45 μm and a diameter of 0.2 μm respectively; the filter with a diameter of 0.2 μm may be a polyethersulfone microporous filter with a diameter of 0.2 μm.

[0124] In an embodiment of the present application, in step (4) of S50, the temperature in the product tank can be controlled to be no higher than 50°C. For example, the temperature in the product tank can be controlled to be 25°C, 30°C, 35°C, 40°C, 45°C or 50°C.

[0125] In an embodiment of the present application, in step (1) of S60, the secondary sterilization filtration may include filtering the drug solution in the product tank with nitrogen, and sterilizing and filtering the filtered drug solution using a polyethersulfone sterilization filter with a diameter of 0.2 μm.

[0126] In an embodiment of the present application, step (1) of S60 may further include depyrogenating and cooling the medicine container, and the depyrogenating temperature may be 330°C to 340°C, for example, 330°C, 332°C, 334°C, 336°C, 338°C or 340°C; the cooling may include a first stage of cooling at a temperature ≤50°C and a second stage of cooling at a temperature ≤35°C.

[0127] In an embodiment of the present application, in step (2) of S60, the number of vacuuming and nitrogen filling may be no less than 5 times, for example, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times or 15 times.

[0128] In an embodiment of the present application, the medicine container may be a bottle that meets pharmaceutical requirements, a double-chamber bag, a multi-chamber bag packaging, or a multi-layer co-extruded infusion film (bag) and other suitable packaging, for example, an infusion bottle, an infusion bag, etc.

[0129] In an embodiment of the present application, in step S70, the terminal sterilization temperature requires the F0 value to be greater than 8. For example, the F0 value may be required to be 12≤F0≤23. For another example, the F0 value may be required to be 12, 14, 16, 18, 20, 22 or 23.

[0130] In an embodiment of the present application, in step S70, the headspace residual oxygen value in the medicine container is required to be no higher than 1%, for example, it can be 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%.

[0131] In an embodiment of the present application, the preparation method may include:

[0132] S10: Cleaning and sterilization of auxiliary items for weighing (weighing equipment); including:

[0133] Dry heat sterilization is controlled at 248°C and the sterilization time is 45 minutes; wet heat sterilization is used for powder mixing tanks and other tanks, with the sterilization temperature not lower than 121°C and the sterilization time being 30 minutes;

[0134] S20: Cleaning and sterilization of filling auxiliary supplies (filling station equipment); including:

[0135] The moist heat sterilization temperature is not less than 121°C, and the sterilization time is 30 minutes. The sterilization temperature of the preparation system is 122°C, and the sterilization time is 30 minutes;

[0136] S30: Weighing and deoxygenation of APIs; including:

[0137] (1) Weighing the API;

[0138] (2) API mixing: The weighed APIs are transported to a sealed powder mixing tank by vacuum extraction and mixed in the powder mixing tank;

[0139] (3) Deoxygenation of API: The powder tank is repeatedly vacuumed and nitrogen filled to remove oxygen-containing gas (e.g., air, etc.) entrained in the gaps between API particles, and then set aside;

[0140] (4) Transfer and feeding of API: transfer to the next process and feed through a closed pipeline (e.g., DEC vacuum suction system);

[0141] (5) Rinse all weighing auxiliary supplies after transferring the API;

[0142] S40: Deoxygenation of water for injection; including:

[0143] (1) After the water for injection is prepared, the dissolved oxygen in the water for injection is removed through a degassing tower; the temperature of the degassing tower shall not be lower than 121°C;

[0144] (2) The deoxygenated water for injection is kept ready for use or transported to a storage tank through a closed pipeline. The storage tank and the liquid preparation tank are kept pressurized with nitrogen for a long time; the nitrogen pressure is not less than 0.2 bar;

[0145] S50: Preparation of drug solution; including:

[0146] (1) System deoxygenation: Vacuum the liquid dispensing tank and fill it with nitrogen for 30 minutes to deoxygenate until the residual oxygen value is ≤1000ppm (if the residual oxygen value in the headspace of the liquid dispensing tank and buffer tank is higher than 1000ppm after 30 minutes, continue to deoxygenate until it is qualified);

[0147] (2) Delivery of water for injection: Under nitrogen protection, the water for injection to be used in step (2) of S40 or the water for injection in the storage tank is delivered to the deoxygenated liquid preparation tank through a closed pipeline. The amount of water for injection added here can be about 72% of the prescribed amount. The temperature of the water for injection is 60±5° C. The dissolved oxygen of the water for injection is less than 40 ppb by evacuating and nitrogen filling the preparation system (confirming that the dissolved oxygen value of the water for injection in the liquid preparation tank does not increase and is less than 40 ppb under stirring);

[0148] (3) dissolving the API: connecting the discharge port of the liquid preparation tank and the discharge port of the powder mixing tank to the two feed ports of a three-way mixer, and connecting the discharge port of the three-way mixer to the feed port of the liquid preparation tank through a nitrogen-filled pipeline; discharging the API treated in step (2) of S30 and the water for injection in the liquid preparation tank through the discharge port of the powder tank and the discharge port of the liquid preparation tank in a certain proportion, respectively, and mixing and dissolving them in the three-way mixer, and returning the dissolved liquid to the liquid preparation tank through the discharge port of the three-way mixer; The liquid is circulated multiple times between the liquid preparation tank and the three-way mixer, and the API in the powder mixing tank is continuously transported to the three-way mixer during the multiple circulations of the liquid until the API in the mixing tank is completely transported to the three-way mixer; the rotation speed of the three-way mixer is 45 Hz (about 300 revolutions per minute), and the dissolution temperature of the API is 60±5° C. The powder mixing tank is rinsed with injection water twice, and about 50 kg of water is added each time, and the injection water after rinsing is sent to the liquid preparation tank through the circulating dissolution pipeline;

[0149] (4) Preparation of drug solution, primary sterilization filtration and volume adjustment: vacuum the product tank and fill it with nitrogen for deoxygenation for 30 minutes until the residual oxygen value in the headspace of the product tank is ≤1000ppm (if the residual oxygen value in the headspace is higher than 1000ppm after 30 minutes, continue deoxygenation until it is qualified); start the circulation cooling program of the liquid preparation tank, and the drug solution in the liquid preparation tank is circulated and cooled to 40±5℃ through the heat exchanger; the drug solution in the liquid preparation tank is sterilized and filtered once through the product delivery pump (using a filter with a diameter of 0.45μm and a polyethersulfone microporous filter with a diameter of 0.2μm for sterilization filtration, respectively) to the product tank that has been filled with nitrogen to replace oxygen. After the filtration is completed, use nitrogen Purge the residual liquid medicine in the pipeline to the product tank, and then use injection water to clean the liquid preparation tank, pipeline, and filter element three times. The water consumption each time can be about 40 to 60 kg (if there is excess water for the third cleaning, it will be automatically adjusted according to the final fixed capacity of the system). The injection water after cleaning is pressed into the product tank (as a liquid storage tank to store liquid medicine) with nitrogen and fixed to ±2% of the specified amount; pass cooling water through the jacket of the fixed-volume product tank to cool it down (the temperature displayed in the product tank is not less than 50°C), and stir it, with a speed of 45Hz (about 300 rpm), and the nitrogen bubbling pressure is not less than 0.2bar, which continues until the filling is completed;

[0150] S60 filling; including:

[0151] (1) Secondary sterilization filtration and filling:

[0152] 1) Sterilization and filtration before filling: Start the product filling buffer program of the preparation system, set the filtration pressure ≤ 1.5 bar (according to the set pressure limit, the system automatically compensates the pressure to keep the filtration pressure constant), and the system runs; use nitrogen to filter the liquid in the product tank, and transport it to the metering storage tank of the filling machine through a polyethersulfone terminal sterilization filter with a diameter of 0.2μm;

[0153] 2) The metering tank is equipped with an online liquid level probe to monitor the amount added. When the liquid level is high, the valve of the metering tank is automatically closed; when the liquid level of the metering tank is low during the filling process, the valve automatically opens to add liquid medicine until the filling is completed;

[0154] 3) Cleaning of medicine bottles: rinse the medicine bottles with water for injection filtered by a microporous filter with a diameter of 0.2 μm and a fully automatic bottle washing machine, and blow away the residual water with clean compressed air. After the inner wall is washed four times with water for injection and blown dry with clean compressed air four times, enter the tunnel oven for sterilization and pyrogen removal;

[0155] 4) Pyrogen removal of medicine bottles: Sterilization and pyrogen removal are carried out in a tunnel-type hot air circulation oven, where the temperature is 330°C to 340°C, the temperature of cooling section 1 is ≤50°C, and the temperature of cooling section 2 is ≤35°C; the speed of the tunnel oven conveyor belt is controlled to be ≤203mm / min;

[0156] 5) Rubber stopper cleaning and sterilization: The rubber stopper is cleaned and sterilized using a fully automatic rubber stopper cleaning and sterilization machine, wherein the rubber stopper is roughly washed 3 times with injection water and finely washed once, and the compressed air pressure during cleaning is ≥4 bar; pure steam wet heat sterilization is performed for 30 minutes, and the sterilization temperature range is 122°C to 124°C;

[0157] 6) Transfer and loading of aluminum caps: The aluminum caps are transported to the capping room in a sealed manner and added to the hopper of the capping machine. The aluminum caps can be transferred in batches;

[0158] (2) Filling and stoppering process

[0159] 1) Adding rubber stoppers: transporting the sterilized rubber stoppers and adding them into the rubber stopper hopper;

[0160] 2) Adjust the filling volume; through the time pressure valve control, the measured filling volume should be ≥252ml;

[0161] 3) Adjust the residual oxygen content in the headspace: adjust the vacuum and nitrogen filling parameters until the residual oxygen content in the air above the vial after filling and stoppering is ≤1.0%;

[0162] 4) Filling and stoppering: including three steps of nitrogen filling and deoxygenation protection: nitrogen filling of empty bottles before filling → filling with liquid medicine and nitrogen filling simultaneously → vacuuming before stoppering - nitrogen filling multiple times, and finally stoppering and sealing under C-level laminar flow and A-level air supply;

[0163] (3) Capping process (including vacuum leak detection and torque measurement): Take each capped medicine bottle and first test the vacuum leakage value (≤3.84mbar), then test the torque (≥9.25kgf·cm, the torque limit of the initial inspection before production). If qualified, start the formal capping process;

[0164] S70: terminal sterilization, light inspection, headspace residual oxygen detection and packaging; including:

[0165] (1) Terminal sterilization process: sterilization is carried out by overkill method, equipment setting parameters: minimum sterilization temperature: 121.1℃, maximum sterilization temperature: 123.0℃, control temperature: 122℃; minimum F0 calculation temperature: 110.0℃; control F0 value: 12<F0≤23; after sterilization, the temperature is lowered to 40℃ and the product is taken out of the cabinet;

[0166] (2) Light inspection process: Products that fail to meet the inspection standards for visible foreign matter, abnormal filling volume, empty bottles, exposed stoppers, neck roll marks, and abnormal curling will be removed;

[0167] (3) Headspace residual oxygen detection process: laser residual oxygen instrument non-destructively checks the headspace residual oxygen of the pharmaceutical bottle, and the qualified standard is: headspace residual oxygen ≤ 1.0%;

[0168] (4) Packaging process: Qualified products are labeled, packed and put into storage.

[0169] The following examples will illustrate the technical solution of the present application in more detail, but are not intended to limit the present application.

[0170] (1) Evaluation of raw material dissolution temperature and dissolution time:

[0171] 1.1. Dissolution temperature: Taking thermally unstable alanyl-glutamine as the target of investigation, whether the impurities generated by alanyl-glutamine at different temperatures exceed the predetermined standard is used as the basis for determining the appropriate dissolution temperature. The data are shown in Table 1.

[0172] Table 1 Effect of dissolution temperature on impurities produced by alanyl-glutamine

[0173]

[0174] Conclusion: The dissolution temperature of 55°C to 65°C and the dissolution time of less than 2 hours are suitable for the stability of alanyl-glutamine.

[0175] 1.2. Dissolution temperature and time of raw material mixture: First, add the same volume (90% of the total liquid volume of water for injection) of water for injection at different temperatures, with the upper limit of the temperature being the circulating temperature of water for injection at 70°C, and then add the prescribed amount (i.e., per 100 ml, containing: aspartic acid 4.00 g, threonine 4.32 g, serine 5.00 g, glutamic acid 1.60 g, proline 5.00 g, glycine 3.20 g, alanine 0.60 g, N(2)-L-alanyl-L-glutamyl A solid mixture of 20 kinds of amino acids (18.05 g, 4-amino-3-amino-2-nitropropene 18.05 g, valine 9.50 g, methionine 3.20 g, isoleucine 7.50 g, leucine 13.00 g, phenylalanine 3.75 g, lysine acetate 7.05 g, histidine 2.60 g, arginine 8.60 g, acetylcysteine ​​1.21 g, acetyltyrosine 0.92 g, tryptophan 1.08 g, taurine 1.00 g) was weighed and the dissolution time was observed under the same stirring speed. The data are shown in Table 2.

[0176] Table 2 Dissolution rate test of 20 kinds of amino acids mixed feed

[0177] Dissolving temperature 65℃ to 70℃ 60℃ to 65℃ 55℃ to 60℃ Time required for complete dissolution 9min 11min 25min

[0178] Conclusion: Considering the dissolution temperature and dissolution time comprehensively, a dissolution temperature of 60℃ to 65℃ is more appropriate.

[0179] (2) Experimental evaluation of the anaerobic control system of production equipment:

[0180] 2.1. Evaluation of the anaerobic control system in the preparation process

[0181] 2.1.1. Evaluation of the anaerobic airtightness of the preparation tank: The prepared amount of water for injection was pumped into the preparation tank. After deoxygenation treatment, the dissolved oxygen value was measured to be 34.2ppb. After a period of time, the dissolved oxygen value did not increase, indicating that the anaerobic airtightness of the preparation tank was very good and met the expected oxygen control requirements.

[0182] 2.1.2. Evaluation of deoxygenation in the powder mixing tank: During the mixing process, the residual oxygen value did not exceed 1000ppm, indicating that the deoxygenation preparation of the powder mixing tank was in line with expectations.

[0183] 2.1.3. Evaluation of the anaerobic airtightness of the product tank: The dissolved oxygen content of the diluted fixed volume solution in the product tank for 72 hours did not exceed 40ppb, indicating that the anaerobic airtightness of the product tank was very good and met the expected oxygen control requirements.

[0184] 2.2. Evaluation of anaerobic control in the filling and plugging process: The effects of "vacuuming-nitrogen filling" 2 to 8 times after filling were compared to test the residual oxygen in the headspace. The test data showed that the average values ​​of 3 to 7 times were basically the same, so "vacuuming-nitrogen filling" should be performed at least 3 times; since the filling speed of "vacuuming-nitrogen filling" 5 times or less is 53 bottles / min, "vacuuming-nitrogen filling" more than 5 times will reduce the filling by 1 bottle per minute for each additional time. Therefore, "vacuuming-nitrogen filling" 5 times has become the optimal number of choices that have excellent deoxygenation effect, sufficient frequency coefficient and do not affect the filling speed. For the actual liquid filling of the amino acid injection described in this application, the optimal choice of the number of "vacuuming-nitrogen filling" is 5 times. In production, the number of "vacuuming-nitrogen filling" is 5 times for continuous filling. The test results of the finished product show that the maximum residual oxygen in the headspace of the sample after filling is 0.36% (the average value is 0.25%), which is in line with the expectation of less than 1%.

[0185] (3) Terminal sterilization temperature evaluation:

[0186] The impurity L-pyroglutamyl-L-alanine produced by the heat-sensitive raw material alanylglutamine was used as the evaluation index, and other impurities were taken into consideration, and the F0 value (standard sterilization time) greater than 12 was used as the lower limit, and the upper limit of the F0 value was investigated. The results are shown in Table 3.

[0187] Table 3 Effect of F0 on impurities produced by alanyl-glutamine

[0188]

[0189] Conclusion: When the F0 value reaches 26.3, L-pyroglutamyl-L-alanine is close to the limit standard, so the preferred F0 value is 12<F0≤23.

[0190] The detection methods used in the quality study of the compound amino acid injection in the examples of this application all use the methods described in the General Rules of Part 4 of the 2020 edition of the Chinese Pharmacopoeia.

[0191] In Example 1 and Comparative Example 1, in the production process of using activated carbon for decolorization, Example 1 does not add a sulfite antioxidant, and Comparative Example 1 adds a sulfite antioxidant; the purpose is to compare the effects of sulfite antioxidants and no sulfite antioxidants on the stability of the compound amino acid injection of the examples of the present application. The results are shown in Stability Study A.

[0192] In Example 2 and Comparative Example 2, in the formula without adding sulfite antioxidant, the effects of using activated carbon and not using activated carbon in the preparation process on the stability and transmittance of the compound amino acid injection of the present application example were compared. The results are shown in Stability Study B.

[0193] Example 1

[0194] The formula of the compound amino acid injection of this example is shown in Table 4.

[0195] Table 4 Formula of the compound amino acid injection of Example 1

[0196]

[0197]

[0198] Note 1: In the actual production process, the amount of each component = theoretical amount × actual feed coefficient. The actual feed coefficient is slightly greater than 1 to offset the loss of activated carbon adsorption during the preparation process.

[0199] The preparation process of the compound amino acid injection of this embodiment is as follows:

[0200] S10: Cleaning and sterilization of auxiliary items for weighing (weighing equipment); including:

[0201] Dry heat sterilization is controlled at 248°C for 45 minutes; wet heat sterilization is used for powder mixing tanks and other containers, with a sterilization temperature not lower than 121°C and a sterilization time of 30 minutes;

[0202] S20: Cleaning and sterilization of filling auxiliary supplies (filling station equipment); including:

[0203] The moist heat sterilization temperature is not less than 121°C, and the sterilization time is 30 minutes. The sterilization temperature of the preparation system is 122°C, and the sterilization time is 30 minutes;

[0204] S30: Weighing and deoxygenation of APIs; including:

[0205] (1) Weighing the API;

[0206] (2) API mixing: The weighed APIs are transported to a sealed powder mixing tank by vacuum extraction and mixed in the powder mixing tank;

[0207] (3) Deoxygenation of API: The powder tank is repeatedly vacuumed and nitrogen filled to remove oxygen-containing gas (e.g., air, etc.) entrained in the gaps between API particles, and then set aside;

[0208] (4) Transfer and feeding of API: transfer to the next process and feed through a closed pipeline (e.g., DEC vacuum suction system);

[0209] (5) Rinse all weighing auxiliary supplies after transferring the API;

[0210] S40: Deoxygenation of water for injection; including:

[0211] (1) After the water for injection is prepared, the dissolved oxygen in the water for injection is removed through a degassing tower; the temperature of the degassing tower shall not be lower than 121°C;

[0212] (2) The deoxygenated water for injection is kept ready for use or transported to a storage tank through a closed pipeline. The storage tank and the liquid preparation tank are kept pressurized with nitrogen for a long time; the nitrogen pressure is not less than 0.2 bar;

[0213] S50: Preparation of liquid medicine; including:

[0214] (1) System deoxygenation: Vacuum the liquid dispensing tank and fill it with nitrogen for 30 minutes until the residual oxygen value is ≤1000ppm (if the residual oxygen value in the headspace of the liquid dispensing tank is higher than 1000ppm after 30 minutes, continue to deoxygenate until it is qualified);

[0215] (2) Delivery of water for injection: Under nitrogen protection, the water for injection to be used in step (2) of S40 or the water for injection in the storage tank is delivered to the deoxygenated preparation tank through a closed pipeline, wherein the amount of water for injection added is about 65% of the prescribed amount, the temperature of the water for injection is 60±5° C., and the dissolved oxygen of the water for injection is less than 40 ppb by evacuating and nitrogen filling the preparation system (confirming that the dissolved oxygen value of the water for injection in the preparation tank does not increase and is less than 40 ppb under stirring);

[0216] (3) dissolving the API: connecting the discharge port of the liquid preparation tank and the discharge port of the powder mixing tank to the two feed ports of a three-way mixer, and connecting the discharge port of the three-way mixer to the feed port of the liquid preparation tank through a nitrogen-filled pipeline; discharging the API treated in step (2) of S30 and the water for injection in the liquid preparation tank through the discharge port of the powder tank and the discharge port of the liquid preparation tank in a certain proportion, respectively, and mixing and dissolving them in the three-way mixer, and returning the dissolved liquid to the liquid preparation tank through the discharge port of the three-way mixer; The liquid is circulated multiple times between the liquid preparation tank and the three-way mixer, and the API in the powder mixing tank is continuously transported to the three-way mixer during the multiple circulations of the liquid until the API in the mixing tank is completely transported to the three-way mixer; the rotation speed of the three-way mixer is 45 Hz (about 300 revolutions per minute), and the dissolution temperature of the API is 55° C. to 65° C.; the powder mixing tank is rinsed with injection water twice, and about 50 kg of water is added each time, and the injection water after rinsing is sent to the liquid preparation tank through the circulating dissolution pipeline;

[0217] (4) Preparation of drug solution, primary sterilization filtration and volume adjustment: vacuum the product tank and fill it with nitrogen for deoxygenation for 30 minutes until the residual oxygen value in the headspace of the product tank is ≤1000ppm (if the residual oxygen value in the headspace is higher than 1000ppm after 30 minutes, continue deoxygenation until it is qualified); start the circulating cooling program of the liquid preparation tank, and the drug solution in the liquid preparation tank is circulated and cooled to 38℃ to 42℃ through the heat exchanger;

[0218] The activated carbon was deoxygenated in the activated carbon tank for 10 minutes, then added to the liquid preparation tank; stirred for 30 minutes;

[0219] Start the cycle decarbonization program, cycle decarbonization for 10 minutes, and confirm that the clarity of the liquid medicine is qualified; the liquid medicine in the preparation tank is decarbonized by a 3μm titanium rod, finely decarbonized by a filter with a diameter of 0.45μm, and finely filtered by a polyethersulfone microporous filter with a diameter of 0.2μm to a product tank (liquid storage tank) that has been filled with nitrogen to replace oxygen. After the filtration is completed, the residual liquid medicine in the pipeline is purged into the product tank with nitrogen, and then the preparation tank, pipeline, titanium rod and filter element are cleaned three times with injection water. The cleaning water is pressed into the product tank (liquid storage tank) with nitrogen and fixed to the prescribed amount. ; Cool the jacket of the fixed-volume product tank with cooling water (the temperature displayed in the product tank is not less than 50℃), and stir it, the speed is 45Hz (about 300 rpm), and the nitrogen bubbling pressure is 0.2bar, which continues until the filling is completed;

[0220] S60 filling; including:

[0221] (1) Secondary sterilization filtration and filling:

[0222] 1) Sterilization and filtration before filling: Start the product filling buffer program of the preparation system, set the filtration pressure ≤ 1.5 bar (according to the set pressure limit, the system automatically compensates the pressure to keep the filtration pressure constant), and the system runs; use nitrogen to filter the liquid in the product tank, and transport it to the metering storage tank of the filling machine through a polyethersulfone terminal sterilization filter with a diameter of 0.2μm;

[0223] 2) The metering tank is equipped with an online liquid level probe to monitor the amount added. When the liquid level is high, the valve of the metering tank is automatically closed; when the liquid level of the metering tank is low during the filling process, the valve automatically opens to add liquid medicine until the filling is completed;

[0224] 3) Cleaning of medicine bottles: rinse the medicine bottles with water for injection filtered by a microporous filter with a diameter of 0.2 μm and a fully automatic bottle washing machine, and blow away the residual water with clean compressed air. After the inner wall is washed four times with water for injection and blown dry with clean compressed air four times, enter the tunnel oven for sterilization and pyrogen removal;

[0225] 4) Pyrogen removal of medicine bottles: Sterilization and pyrogen removal are carried out in a tunnel-type hot air circulation oven, where the temperature is 330°C to 340°C, the temperature of cooling section 1 is ≤50°C, and the temperature of cooling section 2 is ≤35°C; the speed of the tunnel oven conveyor belt is controlled to be ≤203mm / min;

[0226] 5) Rubber stopper cleaning and sterilization: The rubber stopper is cleaned and sterilized using a fully automatic rubber stopper cleaning and sterilization machine, wherein the rubber stopper is roughly washed 3 times with injection water and finely washed once, and the compressed air pressure during cleaning is ≥4 bar; pure steam wet heat sterilization is performed for 30 minutes, and the sterilization temperature range is 122°C to 124°C;

[0227] 6) Transfer and loading of aluminum caps: The aluminum caps are transported to the capping room in a sealed manner and added to the hopper of the capping machine. The aluminum caps can be transferred in batches;

[0228] (2) Filling and stoppering process

[0229] 1) Adding rubber stoppers: transporting the sterilized rubber stoppers and adding them into the rubber stopper hopper;

[0230] 2) Adjust the filling volume; through the time pressure valve control, the measured filling volume should be ≥252ml (weighing method ≥260g, relative density of the drug solution 1.031g / ml);

[0231] 3) Adjust the residual oxygen content in the headspace: adjust the vacuum and nitrogen filling parameters until the residual oxygen content in the air above the vial after filling and stoppering is ≤1.0%;

[0232] 4) Filling and stoppering: including three steps of nitrogen filling and deoxygenation protection: filling in 250ml borosilicate glass infusion bottles under nitrogen protection, vacuuming and nitrogen filling multiple times before stoppering until the residual oxygen in the headspace is ≤1.0%, then stoppering and capping;

[0233] S70: terminal sterilization, light inspection, headspace residual oxygen detection and packaging; including:

[0234] The overkill method was used for sterilization, and the equipment setting parameters were as follows: minimum sterilization temperature: 121.1°C, maximum sterilization temperature: 123.0°C, control temperature: 122°C; minimum temperature for F0 calculation: 110.0°C;

[0235] Control F0 value: 12<F0≤23;

[0236] After sterilization, the mixture was cooled to obtain the injection solution (batch number: 140327).

[0237] Comparative Example 1

[0238] The formula of the compound amino acid injection of this comparative example is shown in Table 5.

[0239] Table 5 Comparative Example 1 Compound amino acid injection formula

[0240]

[0241]

[0242] Note 1: The amount of each component = theoretical amount × actual feed coefficient. The actual feed coefficient is slightly greater than 1 to offset the loss of activated carbon adsorption during the preparation process.

[0243] The preparation process of the compound amino acid injection of this comparative example is as follows:

[0244] Transfer the API and sodium bisulfite into the powder mixing tank by vacuum extraction;

[0245] The other operating steps were the same as those in Example 1, and finally an injection solution (batch number: 140330) was prepared.

[0246] Stability Study A

[0247] The product of Example 1 (batch number 140327) and the product of Comparative Example 1 (batch number 140330) were subjected to stability study in an intermediate condition test (30±2°C / 65%±5%RH). The results of the physical and chemical indicators are as follows:

[0248] Table 6 Stability data of the product of Example 1 (batch number 140327) under intermediate conditions (30±2°C / 65±5%RH)

[0249]

[0250]

[0251] Table 7 Comparative Example 1 (Batch No. 140330) Product Stability Test Data at Intermediate Conditions (30±2°C / 65±5%RH)

[0252]

[0253]

[0254] As can be seen from the above stability data, after being placed for 12 months under the intermediate condition test (30±2°C / 65±5%RH), the acetylcysteine ​​content of the product of Example 1 (batch number is 140327) slightly decreased from the initial 98.8% to 95.5%; the acetylcysteine ​​content of the product of Comparative Example 1 (batch number is 140330) significantly decreased from the initial 98.1% to 54.9%, which is significantly lower than the lower limit of the usual content standard range of 90.0%-110.0% of pharmaceutical preparations, and the decline is several times greater than that of Example 1.

[0255] And after placing and investigating 12 months in the intermediate condition test (30 ± 2 ℃ / 65 ± 5% RH), for the influence of the impurity generation of alanyl-glutamine, the impurity L-pyroglutamyl-L-Alanine of embodiment 1 (lot number is 140327) product rose to 0.64% by initial 0.38%, and the impurity L-alanyl-L-glutamic acid rose to 0.22% by initial 0.08%, all within the qualified range; The impurity L-pyroglutamyl-L-Alanine of comparative example 1 (lot number is 140330) product rose to 0.85% by initial 0.41%, and the impurity L-alanyl-L-glutamic acid rose to 1.91% by initial 0.16%, and the content of these two impurities after being placed for 9 months and 12 months all exceeded the qualified range.

[0256] The active ingredients acetylcysteine ​​and alanylglutamine, which are easily oxidized, have better stability in the composition of Example 1 not containing sodium bisulfite than in the composition of Comparative Example 1 containing sodium bisulfite.

[0257] Example 2

[0258] The formula of the compound amino acid injection of this example is shown in Table 8.

[0259] Table 8 Formula of the compound amino acid injection of Example 2

[0260]

[0261] The preparation process of the compound amino acid injection of this embodiment is as follows:

[0262] S10: Cleaning and sterilization of auxiliary items for weighing (weighing equipment); including:

[0263] Dry heat sterilization is controlled at 248°C for 45 minutes; wet heat sterilization is used for powder mixing tanks and other containers, with a sterilization temperature not lower than 121°C and a sterilization time of 30 minutes;

[0264] S20: Cleaning and sterilization of filling auxiliary supplies (filling station equipment); including:

[0265] The moist heat sterilization temperature is not less than 121°C, and the sterilization time is 30 minutes. The sterilization temperature of the preparation system is 122°C, and the sterilization time is 30 minutes;

[0266] S30: Weighing and deoxygenation of APIs; including:

[0267] (1) Weighing the API;

[0268] (2) API mixing: The weighed APIs are transported to a sealed powder mixing tank by vacuum extraction and mixed in the powder mixing tank;

[0269] (3) Deoxygenation of API: The powder tank is repeatedly vacuumed and nitrogen filled to remove oxygen-containing gas (e.g., air, etc.) entrained in the gaps between API particles, and then set aside;

[0270] (4) Transfer and feeding of API: transfer to the next process and feed through a closed pipeline (e.g., DEC vacuum suction system);

[0271] (5) Rinse all weighing auxiliary supplies after transferring the API;

[0272] S40: Deoxygenation of water for injection; including:

[0273] (1) After the water for injection is prepared, the dissolved oxygen in the water for injection is removed through a degassing tower; the temperature of the degassing tower shall not be lower than 121°C;

[0274] (2) The deoxygenated water for injection is kept ready for use or transported to a storage tank through a closed pipeline. The storage tank and the liquid preparation tank are kept pressurized with nitrogen for a long time; the nitrogen pressure is not less than 0.2 bar;

[0275] S50: Preparation of drug solution; including:

[0276] (1) System deoxygenation: Vacuum the liquid dispensing tank and fill it with nitrogen for 30 minutes until the residual oxygen value is ≤1000ppm (if the residual oxygen value in the headspace of the liquid dispensing tank is higher than 1000ppm after 30 minutes, continue to deoxygenate until it is qualified);

[0277] (2) Delivery of water for injection: Under nitrogen protection, the water for injection to be used in step (2) of S40 or the water for injection in the storage tank is delivered to the deoxygenated preparation tank through a closed pipeline, wherein the amount of water for injection added is about 65% of the prescribed amount, the temperature of the water for injection is 60±5° C., and the dissolved oxygen of the water for injection is less than 40 ppb by evacuating and nitrogen filling the preparation system (confirming that the dissolved oxygen value of the water for injection in the preparation tank does not increase and is less than 40 ppb under stirring);

[0278] (3) dissolving the API: connecting the discharge port of the liquid preparation tank and the discharge port of the powder mixing tank to the two feed ports of a three-way mixer, and connecting the discharge port of the three-way mixer to the feed port of the liquid preparation tank through a nitrogen-filled pipeline; discharging the API treated in step (2) of S30 and the water for injection in the liquid preparation tank through the discharge port of the powder tank and the discharge port of the liquid preparation tank in a certain proportion, respectively, and mixing and dissolving them in the three-way mixer, and returning the dissolved liquid to the liquid preparation tank through the discharge port of the three-way mixer; The liquid is circulated multiple times between the liquid preparation tank and the three-way mixer, and the API in the powder mixing tank is continuously transported to the three-way mixer during the multiple circulations of the liquid until the API in the mixing tank is completely transported to the three-way mixer; the rotation speed of the three-way mixer is 45 Hz (about 300 revolutions per minute), and the dissolution temperature of the API is 55° C. to 65° C.; the powder mixing tank is rinsed with injection water twice, and about 50 kg of water is added each time, and the injection water after rinsing is sent to the liquid preparation tank through the circulating dissolution pipeline;

[0279] (4) Preparation of drug solution, primary sterilization filtration and volume adjustment: vacuum the product tank and fill it with nitrogen for deoxygenation for 30 minutes until the residual oxygen value in the headspace of the product tank is ≤1000ppm (if the residual oxygen value in the headspace is higher than 1000ppm after 30 minutes, continue deoxygenation until it is qualified); start the circulating cooling program of the liquid preparation tank, and the drug solution in the liquid preparation tank is circulated and cooled to 38℃ to 42℃ through the heat exchanger;

[0280] The product tank (liquid storage tank) is evacuated and filled with nitrogen for deoxygenation for 30 minutes; the liquid in the preparation tank is filtered through a polyethersulfone microporous filter with a diameter of 0.2μm through a product delivery pump to a product tank (liquid storage tank) that has been filled with nitrogen to replace oxygen. After the filtration is completed, the residual liquid in the pipeline is purged into the product tank with nitrogen, and then the preparation tank, pipeline and filter element are cleaned three times with water for injection. The cleaning water is pressed into the product tank (liquid storage tank) with nitrogen and fixed to the prescribed amount; the jacket of the product tank that has been fixed to the volume is cooled by cooling water (the temperature displayed in the product tank is not less than 50℃), and stirred at a speed of 45Hz (about 300 rpm), and the nitrogen bubbling pressure is 0.2bar, which is continued until the filling is completed;

[0281] S60 filling; including:

[0282] (1) Secondary sterilization filtration and filling:

[0283] 1) Sterilization and filtration before filling: Start the product filling buffer program of the preparation system, set the filtration pressure ≤ 1.5 bar (according to the set pressure limit, the system automatically compensates the pressure to keep the filtration pressure constant), and the system is running; use nitrogen to filter the liquid in the product tank, and transport it to the filling machine storage tank through a polyethersulfone terminal sterilization filter with a diameter of 0.2μm;

[0284] 2) The metering tank is equipped with an online liquid level probe to monitor the amount added. When the liquid level is high, the valve of the metering tank is automatically closed; when the liquid level of the metering tank is low during the filling process, the valve automatically opens to add liquid medicine until the filling is completed;

[0285] 3) Cleaning of medicine bottles: rinse the medicine bottles with water for injection filtered by a microporous filter with a diameter of 0.2 μm and a fully automatic bottle washing machine, and blow away the residual water with clean compressed air. After the inner wall is washed four times with water for injection and blown dry with clean compressed air four times, enter the tunnel oven for sterilization and pyrogen removal;

[0286] 4) Pyrogen removal of medicine bottles: Sterilization and pyrogen removal are carried out in a tunnel-type hot air circulation oven, where the temperature is 330°C to 340°C, the temperature of cooling section 1 is ≤50°C, and the temperature of cooling section 2 is ≤35°C; the speed of the tunnel oven conveyor belt is controlled to be ≤203mm / min;

[0287] 5) Rubber stopper cleaning and sterilization: The rubber stopper is cleaned and sterilized using a fully automatic rubber stopper cleaning and sterilization machine, wherein the rubber stopper is roughly washed 3 times with injection water and finely washed once, and the compressed air pressure during cleaning is ≥4 bar; pure steam wet heat sterilization is performed for 30 minutes, and the sterilization temperature range is 122°C to 124°C;

[0288] 6) Transfer and loading of aluminum caps: The aluminum caps are transported to the capping room in a sealed manner and added to the hopper of the capping machine. The aluminum caps can be transferred in batches;

[0289] (2) Filling and stoppering process

[0290] 1) Adding rubber stoppers: transporting the sterilized rubber stoppers and adding them into the rubber stopper hopper;

[0291] 2) Adjust the filling volume; through the time pressure valve control, the measured filling volume should be ≥252ml (weighing method ≥260g, relative density of the drug solution 1.031g / ml);

[0292] 3) Adjust the residual oxygen content in the headspace: adjust the vacuum and nitrogen filling parameters until the residual oxygen content in the air above the vial after filling and stoppering is ≤1.0%;

[0293] 4) Filling and stoppering: including three steps of nitrogen filling and deoxygenation protection: filling in 250ml borosilicate glass infusion bottles under nitrogen protection, vacuuming and nitrogen filling multiple times before stoppering until the residual oxygen in the headspace is ≤1.0%, then stoppering and capping;

[0294] The over-killing method was used for sterilization, and the equipment setting parameters were: minimum sterilization temperature: 121.1°C, maximum sterilization temperature: 123.0°C, control temperature: 122°C; F0 calculated minimum temperature: 110.0°C.

[0295] The control F0 value is: 12<F0≤23.

[0296] After sterilization, the solution was cooled to obtain the injection (batch number: 2005001).

[0297] Comparative Example 2

[0298] The formula of the compound amino acid injection of this comparative example is shown in Table 9.

[0299] Table 9 Comparative Example 2 Compound Amino Acid Injection Formula

[0300]

[0301]

[0302] The preparation method of the compound amino acid injection of Comparative Example 2 is the same as the preparation method of the compound amino acid injection of Example 1.

[0303] The prepared injection (batch number is 2004004).

[0304] Stability Study B

[0305] The product of Example 2 (batch number 2005001) and the product of Comparative Example 2 (batch number 2004004) were subjected to a long-term stability test (test conditions 25±2°C / 60%±5%RH). The results of the physical and chemical indicators are as follows:

[0306] Table 10 Stability data of Example 2 product (batch number 2005001) at 25±2°C / 60%±5%RH

[0307]

[0308]

[0309]

[0310] Table 11 Stability data of the product of comparative example 2 (batch number 2004004) at 25±2°C / 60%±5%RH

[0311]

[0312]

[0313]

[0314] It can be seen from the above stability data that after being placed under 25±2°C / 60%±5%RH for 18 months, the acetylcysteine ​​content of the product of Example 2 (batch number 2005001) slightly decreased from the initial 99.0% to 97.1%, and the stability was good. Moreover, there was no difference in light transmittance and other indicators between the product of Example 2 and the product of Comparative Example 2 (batch number 2004004).

[0315] In summary, it is surprising that the well-known active ingredient "acetylcysteine" which is easily oxidized has better stability in the compound amino acid injection of Examples 1 and 2 of the present application that does not contain sodium bisulfite than in the composition containing sodium bisulfite.

[0316] Example 3

[0317] A multicenter, randomized, double-blind, positive drug-controlled study was conducted using the compound amino acid injection prepared in Example 1 of the present application and the 8.5% compound amino acid injection (18AA-II) commonly used in clinical practice. 270 patients who underwent elective gastric, small intestine, and colorectal resection (including laparoscopy) and required parenteral nutrition after surgery were selected to compare the effectiveness and safety of the compound amino acid injection described in the present application (study group) and the 8.5% compound amino acid injection (control group) commonly used in clinical practice on parenteral nutrition (PN) support for postoperative patients. Give an equal amount of nitrogen (0.20g / kg·d), equivalent to 1.25g / kg·d of amino acids; non-protein calories (20kcal / kg·d), of which the fat emulsion dose is 1.0g / kg·d, and the remaining calories are provided by glucose. During the study, patients were given appropriate amounts of water-soluble vitamins, fat-soluble vitamins, trace elements, etc. as needed. Starting from the first day after surgery, once a day for 6 consecutive days. The route of administration was designed to be peripheral and central venous administration, but in the study, the vast majority (more than 90%) adopted central venous administration. The baseline of the two groups was balanced before surgery.

[0318] The results of the study showed that:

[0319] (1) After surgery, the glutamine concentrations of both the study group and the control group decreased. On the third and seventh days after surgery, the decrease in the study group was significantly less than that in the control group. On the seventh day after surgery, the study group had recovered and exceeded the level on the first day after surgery, while the control group had not recovered to the level on the first day after surgery. There were significant differences between the two groups. As shown in Table 12 and Figure 1 to Figure 2 As shown:

[0320] Table 12 Analysis of glutamine changes from D0 to D7 after surgery in patients undergoing open and laparoscopic surgery (umol / L)

[0321]

[0322] (2) In terms of hospitalization time, the hospitalization time of the study group was 9.05±2.30 days, and that of the control group was 12.47±10.97 days, showing a significant difference between the two groups. As shown in Table 13:

[0323] Table 13 Comparison of hospital stay between the study group and the control group

[0324]

[0325] (3) There was no difference in the frequency and severity of adverse reactions between the study group and the control group, and no serious or unexpected adverse reactions were reported. In the clinical study: There were 134 cases in the control group, of which 509 adverse events occurred, with an incidence of 91.79%. The adverse events of grade 1 / mild, grade 2 / moderate, and grade 3 / severe were 49 (36.57), 56 (41.79), and 18 (13.43) cases (%), respectively. Among the 18 cases of grade 3, 11 cases were elevated γ-transpeptidase and 1 case was elevated alanine aminotransferase; There were 135 cases in the study group, of which 112 cases had 437 adverse events, with an incidence of 82.35%; The number of cases of elevated leukocytes in the study group was more than that in the control group, all of which were grade I. There were no differences in other indicators between the groups. There were no differences between the groups in adverse events related to the study drug.

[0326] It is unexpectedly seen from the results of Example 3 that the compound amino acid injection described in the present application has better performance in nutritional therapy and nutritional support than the commonly used compound amino acid injection in clinical practice, and is suitable for treating or improving amino acid metabolism disorders and hypoproteinemia caused by trauma, burns, various surgical operations, severe infections or intestinal flora translocation, and improving the body's immune function and intestinal function.

[0327] Although the embodiments disclosed in this application are as above, the contents described are only embodiments adopted to facilitate understanding of this application and are not intended to limit this application. Any technician in the field to which this application belongs can make any modifications and changes in the form and details of implementation without departing from the spirit and scope disclosed in this application, but the scope of protection of this application shall still be based on the scope defined in the attached claims.

Claims

1. A method for preparing a compound amino acid injection, characterized in that: The compound amino acid injection comprises an amino acid composition and does not include a sulfite antioxidant, wherein the amino acid composition comprises the following active substances in a weight ratio, each active substance being present in the form of original form, a pharmaceutically acceptable salt or a short peptide: The method comprises: S10: Cleaning and sterilization of auxiliary items for weighing; S20: Cleaning and sterilization of auxiliary supplies for filling; S30: Weighing and deoxygenation of APIs; including: (1) Weighing the API; (2) Place all the weighed APIs into a sealed powder mixing tank; (3) performing multiple vacuum evacuations and nitrogen fillings on the powder mixing tank to remove oxygen-containing gas entrained between the API particles; S40: Deoxygenation of water for injection; including: (1) After the water for injection is prepared, the dissolved oxygen in the water for injection is removed; (2) The deoxygenated water for injection is kept ready for use or transported to a storage tank through a closed pipeline, and the storage tank is continuously pressurized with nitrogen; S50: Preparation of liquid medicine; including: (1) Deoxygenate the liquid dispensing tank; (2) under nitrogen protection, the water for injection to be used in step (2) of S40 or the water for injection in the storage tank is transported through a closed pipeline to a liquid preparation tank maintained under nitrogen pressure; (3) The API processed in step (3) of S30 and the water for injection in the liquid preparation tank are respectively discharged through the discharge port of the powder mixing tank and the discharge port of the liquid preparation tank in a certain proportion, and mixed and dissolved in a mixer, and the dissolved liquid is returned to the liquid preparation tank through the discharge port of the mixer; the liquid is circulated multiple times between the liquid preparation tank and the mixer, and the API in the powder mixing tank is continuously transported to the mixer during the multiple circulations of the liquid until the API in the mixing tank is completely transported to the mixer; (4) Cooling and sterilizing the liquid medicine prepared in step (3) of S50, and transferring the cooled and filtered liquid medicine to a deoxygenated product tank under nitrogen protection, continuously passing nitrogen into the product tank containing the liquid medicine and controlling the temperature in the product tank; S60: Filling; including: (1) performing secondary sterilization filtration on the drug liquid in the product tank, and filling it into a drug container that has been depyrogenated and cooled under nitrogen protection, pre-filling the drug container with nitrogen before filling to expel oxygen in the drug container, and filling with nitrogen during filling to reduce oxygen intrusion; (2) Vacuuming and nitrogen filling the medicine container with liquid medicine several times and then sealing it with a plug; (3) rolling the cap on the sealed pharmaceutical container; S70: terminal sterilization, lamp inspection, headspace residual oxygen detection and packaging to obtain the compound amino acid injection. Among them, the residual oxygen value of the powder mixing tank after vacuuming and nitrogen filling, the liquid preparation tank maintained by nitrogen pressure, and the product tank after deoxygenation is ≤1000ppm; Wherein, the dissolved oxygen content in the injection water obtained in step (1) of S40 is less than 40 ppb; Wherein, the temperature for dissolving the API in step (3) of S50 is 55° C. to 65° C.; Wherein, in step (4) of S50, the liquid medicine prepared in step (3) of S50 is cooled to 35° C. to 45° C.; Wherein, in step (4) of S50, the temperature in the product tank is controlled to be no higher than 50°C; Wherein, step (1) of S60 further includes depyrogenating and cooling the medicine bottle, wherein the depyrogenating temperature is 330° C. to 340° C., and the cooling includes a first stage cooling at a temperature of ≤50° C. and a second stage cooling at a temperature of ≤35° C.; Wherein, in step (2) of S60, the number of times of vacuuming and nitrogen filling is not less than 5 times; Wherein, in step S70, the terminal sterilization temperature requires the F0 value to be greater than 8; Wherein, in step S70, the headspace residual oxygen value in the medicine container is required to be no higher than 1%.

2. The preparation method according to claim 1, wherein In step (4) of S50, the primary sterilization filtration includes filtering using filters with a diameter of 0.45 μm and a diameter of 0.2 μm respectively.

3. The preparation method according to claim 1, wherein In step (1) of S60, the secondary sterilization filtration includes filtering the drug solution in the product tank with nitrogen, and sterilizing and filtering the filtered drug solution with a polyethersulfone sterilization filter having a diameter of 0.2 μm.

4. The preparation method according to claim 1, wherein In step S70, the terminal sterilization temperature requires the F0 value to be 12≤F0≤23.

5. The preparation method according to claim 1, wherein the compound amino acid injection does not include an antioxidant.

6. A compound amino acid injection, prepared according to the method according to any one of claims 1 to 5, wherein the formula of the compound amino acid injection is: in, In the actual production process, the amount of each component = theoretical amount × actual feed coefficient.

7. A compound amino acid injection, prepared according to the method according to any one of claims 1 to 5, wherein the formula of the compound amino acid injection is: in, In the actual production process, the amount of each component = theoretical amount × actual feed coefficient.

8. Use of the compound amino acid injection according to claim 6 or 7 in the preparation of a medicament for treating or improving hypoproteinemia.

9. The use according to claim 8, wherein the hypoproteinemia is caused by trauma, burns, various surgical operations, severe infection or intestinal flora translocation.

10. The use according to claim 8, wherein the drug improves the body's immune function and intestinal function.

11. The use according to claim 9, wherein the drug improves the body's immune function and intestinal function.

12. Use of the compound amino acid injection according to claim 6 or 7 in the preparation of a medicament for treating or improving amino acid metabolic disorders caused by trauma, burns, various surgical operations, severe infections or intestinal flora translocation.

13. The use according to claim 12, wherein the drug improves the body's immune function and intestinal function.

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