Trace element compositions, methods for their preparation and uses

Abstract

A stable trace element composition, its preparation method, and its use are disclosed. The trace element composition comprises a main material and auxiliary materials. The main material comprises pharmaceutical organic acid salts of iron, zinc, copper, and manganese, pharmaceutical salts of fluorine, iodine, selenium, and molybdenum, and a pharmaceutical organic or inorganic salt of chromium. The auxiliary materials comprise water for injection and a pH adjuster, which is an inorganic acid. The pH value of the trace element composition is 2.0 to 3.5. The amount of each trace element per 10 mL of the trace element composition is (unit: μmol): 17.8 to 21.5 μmol iron, 40 to 100 μmol zinc, 4.7 to 9.6 μmol copper, 1 μmol manganese, 25 to 60 μmol fluorine, 0.5 to 1.2 μmol iodine, 0.75 to 1.27 μmol selenium, 0.2 to 0.26 μmol molybdenum, and 0.2 to 0.4 μmol chromium.

Description

[Technical Field]

[0001] The present application relates to the field of pharmaceutical formulations, in particular to trace element compositions, their preparation methods and their uses. [Background technology]

[0002] Trace elements are essential micronutrients for living organisms, serving as components of certain enzymes, hormones, and other substances in the body. Trace elements cannot be synthesized by the human body and must be replenished daily. Generally, trace elements can be replenished through diet. Demand may increase in some special physiological and pathological conditions, such as pregnancy, or in cases of excessive catabolism (e.g., surgery, major trauma, burns), insufficient supply, abnormal loss, or malabsorption (e.g., short bowel disease or Crohn's disease). To ensure normal physiological function, trace element preparations are commonly required as supplements. Patients requiring parenteral nutrition, especially those with poor digestion and absorption or who are unable to take oral intake, require appropriate amounts of various trace elements. Generally, when used in patients with nutritional deficiencies or increased nutritional losses, commercially available multi-trace element parenteral nutrition preparations are the basic choice, and individual preparations of single trace element preparations are rarely used. In addition to the limited number of commercially available single-element preparations, the large number of trace elements required makes the clinical preparation of single-element preparations complex, prone to preparation variability and the risk of microbial contamination during the preparation process.

[0003] Currently, most trace element products are compound preparations, such as Multitrace-4 (containing the trace elements zinc, copper, chromium, and manganese) and Multitrace-5 (containing the trace elements zinc, copper, chromium, manganese, and selenium) in the North American market, Decan (containing 10 trace elements: iron, zinc, copper, manganese, fluorine, cobalt, iodine, selenium, molybdenum, and chromium) in the European market, Additrace, Tracutil, and Addaven (containing nine trace elements: iron, zinc, copper, manganese, fluorine, iodine, selenium, molybdenum, and chromium, respectively) in the European market, and Multitrace Element Injection and Multitrace Element Injection (II) in the Chinese market. The former contains 10 trace elements like Decan in the European market, while the latter (such as Addamel) contains nine trace elements like Additrace in the European market, although with slightly different supplementary ingredients. Pediatric trace element injections include Neotrace-4 and Peditrace, and a multi-trace element injection (I) with the same composition as Peditrace is available on the domestic market.

[0004] The compositions of multi-trace element injections available on the major markets are shown in Table 1.

[0005] [Table 1]

[0006] In the table, the content refers to the dosage in unit specifications (count or bottle); "_" indicates not included; pH values ​​are according to the product instructions.

[0007] Gluconic acid lactone, a pH adjuster and stabilizer in DECAN products, can undergo hydrolysis in solution to produce gluconic acid, which can then be cyclized to form six-membered cyclic gluconic acid delta-lactone and five-membered cyclic gluconic acid gamma-lactone (Chromatography, Volume 8, Issue 4, 1990, Study on Hydrolysis of D-gluconic Acid Delta-Lactone Using High-Performance Liquid Chromatography). A literature study (Nature, August 24, 1963, pp. 765-767) also showed that gluconic acid delta-lactone, gluconic acid gamma-lactone, and gluconic acid can be interconverted in solution. The interconversion and equilibrium between gluconic acid lactone and its solution chelates relevant trace elements in DECAN products, acting as a stabilizer. Through process testing and testing, it was determined that in addition to the main ingredient, gluconic acid (gluconic acid lactone), the amount of gluconic acid ester added as an auxiliary material in DECAN products is approximately 2 mg / mL.

[0008] Most multi-trace element formulations consist of various trace element salts and auxiliary materials. Typical components include inorganic and organic salts of trace elements. Because multi-trace element formulations form complex formulations, these formulations must meet relevant requirements for clinical use, such as safety and accurate dosing. Interactions between various trace elements and ions can easily lead to redox reactions, resulting in precipitation and discoloration of the solution. In addition, differences in the preparation process can also lead to stability issues. Currently, various techniques are used to resolve the stability issue, including oversupply, adding stabilizers, producing lyophilized formulations, and adding activated carbon. Furthermore, techniques that include stabilizers such as taurine, gluconic acid, xylitol, sorbitol, lactobionic acid, glycine, theanine, lysine, arginine, cysteine, methionine, trehalose, and erythritol are commonly used. The pH of injectable formulations with different formulations varies. According to the prior art, injections containing trace elements in the form of inorganic salts generally have a relatively low pH, typically between 2.0 and 2.6, while injections containing trace elements in the form of organic salts generally have a relatively high pH, ​​typically between 3.5 and 5.0, and at most 4.0 to 4.5. Furthermore, in products with a high concentration of organic salts, the pH adjuster is typically an organic acid such as gluconic acid (gluconic acid lactone).

[0009] Contents of the invention The present application provides a trace element composition, a preparation method thereof and its use, and aims to provide a trace element formulation with a simple composition and good stability.

[0010] In a first aspect of the present application, there is provided a trace element composition, comprising a main material and auxiliary materials, wherein the main material is a pharmaceutical organic acid salt of iron, a pharmaceutical organic acid salt of zinc, a pharmaceutical organic acid salt of copper, a pharmaceutical organic acid salt of manganese, a pharmaceutical salt of fluorine, a pharmaceutical salt of iodine, a pharmaceutical salt of selenium, a pharmaceutical salt of molybdenum, a pharmaceutical organic acid salt of chromium, or a pharmaceutical inorganic salt; and the auxiliary materials are water for injection and a pH adjuster, wherein the pH adjuster is an inorganic acid and an optional inorganic base; The trace element composition has a pH value of 2.0 to 3.5, and the trace elements in the main material are contained in the following amounts per 10 mL of the trace element composition: 17.8 to 21.5 μmol of iron, 40 to 100 μmol of zinc, 4.7 to 9.6 μmol of copper, 1 μmol of manganese, 25 to 60 μmol of fluorine, 0.5 to 1.2 μmol of iodine, 0.75 to 1.27 μmol of selenium, 0.2 to 0.26 μmol of molybdenum, and 0.2 to 0.4 μmol of chromium.

[0011] Furthermore, per 10 ml of trace element composition, each trace element in the main material meets the following dosage: 18-21.5 μmol iron, 50-100 μmol zinc, 4.7-6 μmol copper, 1 μmol manganese, 50 μmol fluorine, 0.5-1.0 μmol iodine, 1.0-1.25 μmol selenium, 0.2 μmol molybdenum and 0.2 μmol chromium.

[0012] Furthermore, per 10 ml of trace element composition, each trace element in the main material is present in the following amounts: 20 μmol iron, 77 μmol zinc, 4.7 μmol or 6 μmol copper, 1 μmol manganese, 50 μmol fluorine, 1 μmol iodine, 1 μmol selenium, 0.2 μmol molybdenum and 0.2 μmol chromium.

[0013] Furthermore, the pH value of the composition is 2.6 to 3.2.

[0014] Furthermore, the inorganic acid is hydrochloric acid or sulfuric acid, preferably hydrochloric acid; the inorganic base is sodium hydroxide or potassium hydroxide, preferably sodium hydroxide.

[0015] Furthermore, the organic acid salt of iron element is selected from any one of ferrous gluconate, ferrous L-aspartate, ferrous DL-aspartate, and ferrous fumarate; the organic acid salt of zinc element is selected from any one of zinc gluconate, zinc L-aspartate, and zinc DL-aspartate; the organic acid salt of copper element is selected from any one of copper gluconate, copper L-aspartate, and copper DL-aspartate; and the organic acid salt of manganese element is selected from any one of manganese gluconate, manganese L-aspartate, and manganese DL-aspartate. the pharmaceutical salt of elemental fluorine is sodium fluoride or potassium fluoride, the pharmaceutical salt of elemental iodine is sodium iodide or potassium iodide, the pharmaceutical salt of elemental selenium is sodium selenite or selenious acid, the pharmaceutical salt of elemental molybdenum is sodium molybdate or ammonium heptamolybdate, the pharmaceutical inorganic salt of elemental chromium is chromium chloride or chromium sulfate, and / or the organic acid salt of elemental chromium is selected from any one of chromium gluconate, chromium L-aspartate, and chromium DL-aspartate.

[0016] Furthermore, the main materials consist of ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, potassium iodide or sodium iodide, sodium selenite or sodium selenite, sodium molybdate and chromium chloride.

[0017] Furthermore, the trace element composition is an injection; preferably, the inner wall of the packaging material of the injection is made of a non-glass packaging material; more preferably, the inner wall of the packaging material is made of a polypropylene material.

[0018] According to another aspect of the present application, there is provided a method for preparing any of the above-mentioned trace element compositions, comprising the steps of dispersing each trace element salt in water for injection, adjusting the pH to a target value using a pH adjuster, filtering to obtain a filtrate, adding water to the filtrate to make up the total volume, potting, and sterilizing, wherein the sterilization is preferably carried out using a terminal sterilization step, and the F0 value of the terminal sterilization step is 8 or more, preferably 12 or more.

[0019] Additionally, the packaging material that comes into direct contact with the medication during potting is a non-glass packaging material, preferably a polypropylene material.

[0020] Furthermore, the trace element composition is prepared under conditions where it is exposed to an illumination intensity of 500 lux or less, preferably an illumination intensity of 300 lux or less, and more preferably an illumination intensity of 100 lux or less.

[0021] According to another aspect of the present application, there is provided an application of any of the above-mentioned trace element compositions in a medicament for the prevention and treatment of trace element deficiency.

[0022] The present composition is formed by replacing some inorganic salt elements in Addaven products. Surprisingly, the composition formed after this replacement has been found to have better safety, and the formulation has better stability within the proposed pH range without the need for additional stabilizers. The content of some elements (such as iodine) is more stable, eliminating the need for overdose. Furthermore, the trace element composition of the present invention has a significantly reduced product specification compared to Decan, making it more convenient and safe for clinical use.

[0023] DESCRIPTION OF THE DRAWINGS In order to more clearly describe the technical solutions of the embodiments of the present application, the drawings necessary for use in the embodiments of the present application are briefly described below. However, it is obvious to those skilled in the art that the drawings described below are only a part of the embodiments of the present application, and other drawings can be obtained according to the drawings without creative efforts. [Brief explanation of the drawings]

[0024] [Figure 1] FIG. 1 shows photographs of samples of Example 1 with H values ​​of 2.0, 2.2, 2.4, 2.6, 2.9, 3.2, 3.5, 4.0, and 4.3. [Figure 2] FIG. 2 shows photographs of test tubes Nos. 1 to 7 that were thoroughly shaken after adding a sample in an in vitro hemolysis test on rabbit red blood cells in the Examples. [Figure 3] FIG. 3 shows photographs of test tubes Nos. 1 to 7 taken 3 hours after adding the sample in an in vitro hemolysis test on rabbit red blood cells in an example. [Figure 4] FIG. 4 shows photographs of test tubes Nos. 1 to 7 that were thoroughly shaken 3 hours after the addition of the sample in the in vitro hemolysis test for rabbit red blood cells in the Examples. [Figure 5] FIG. 5 shows the results of histopathological examination of the injection sites on the treated side (left photograph) and the control side (right photograph) after 14 days of recovery in animals administered with the control sample. [Figure 6] FIG. 6 shows the histopathological examination of the proximal non-injected site on the treated side (left photograph) and the control side (right photograph) after 14 days of recovery in animals administered the control sample. [Figure 7] FIG. 7 shows the results of histopathological examination of the injection sites of the treated side (left photograph) and the control side (right photograph) in the animals administered with the test sample after 14 days of recovery. [Figure 8] FIG. 8 shows the results of histopathological examination of the proximal non-injected site on the treated side (left photograph) and the control side (right photograph) after 14 days of recovery in animals administered with the test sample. [Figure 9] FIG. 9 shows the curves of changes in plasma copper concentration after administration of each combination drug in the test groups in Table 29. [Figure 10] FIG. 10 shows the curves of changes in plasma manganese concentration after administration of each combination preparation in the test groups in Table 29. [Figure 11] FIG. 11 shows the curves of changes in plasma zinc concentration after administration of each combination preparation in the test groups in Table 29. [Figure 12]FIG. 12 shows the curves of changes in plasma selenium concentration after administration of each combination preparation in the test groups in Table 29. [Figure 13] FIG. 13 shows the white blood cell counts of the model animals before administration of each combination preparation in the test groups in Table 29. [Figure 14] FIG. 14 shows the white blood cell counts of model animals after administration of each combination preparation in the test groups in Table 29. [Figure 15] FIG. 15 shows a typical graph of irritation after tail vein administration of each combination preparation in the test group in Table 29 for four consecutive days. DETAILED DESCRIPTION OF THE INVENTION

[0025] Detailed Description of Specific Embodiments The embodiments of the present application will be described in more detail with reference to the drawings and examples. The detailed description of the following examples and the accompanying drawings is provided to explain the principles of the present application, but is not intended to limit the scope of the present application, i.e., the present application is not limited to the described examples.

[0026] As described in the background art of this application, Addaven products use hydrochloric acid to adjust the pH and xylitol as a stabilizer to improve product stability. During research into this product, it was discovered that while xylitol can improve product stability, significant loss of iodine content occurs after sterilization. To achieve the desired iodine content in the product, excessive iodine must be added. This is due to the insufficient stability of Addaven products. To solve this problem, the applicant made various attempts in terms of pH, the amount of stabilizer added, and the type of stabilizer, but none of them were able to improve the stability of elemental iodine. After experiencing these failures, the applicant broke away from conventional thinking by replacing inorganic salts of iron, zinc, copper, and manganese with organic salts such as gluconate and using inorganic acids instead of organic acids as pH adjusters to adjust the pH. This not only ensures the stability of the resulting composition without excessive iodine supply, but also eliminates the need for stabilizers, further improving the safety and compliance of the resulting trace element composition.

[0027] In an exemplary embodiment of the present application, there is provided a trace element composition, comprising a main material and an auxiliary material, wherein the main material comprises a pharmaceutical organic acid salt of iron element, a pharmaceutical organic acid salt of zinc element, a pharmaceutical organic acid salt of copper element, a pharmaceutical organic acid salt of manganese element, a pharmaceutical salt of fluorine element, a pharmaceutical salt of iodine element, a pharmaceutical salt of selenium element, a pharmaceutical salt of molybdenum element, a pharmaceutical organic acid salt of chromium element, or a pharmaceutical inorganic salt, and the auxiliary material comprises water for injection and a pH adjuster, wherein the pH adjuster is an inorganic acid and optionally The pH value of the trace element composition is 2.0 to 3.5. Per 10 ml of the trace element composition, the amounts of each trace element in the main material are as follows: 17.8 to 21.5 μmol of iron, 40 to 100 μmol of zinc, 4.7 to 9.6 μmol of copper, 1 μmol of manganese, 25 to 60 μmol of fluorine, 0.5 to 1.2 μmol of iodine, 0.75 to 1.27 μmol of selenium, 0.2 to 0.26 μmol of molybdenum, and 0.2 to 0.4 μmol of chromium.

[0028] As mentioned above, after replacing some of the inorganic salt elements in Addaven products, the resulting product was surprisingly safe and stable within the proposed pH range without the need for additional stabilizers. The content of some elements (such as iodine) was more stable, eliminating the need for excessive supply. Furthermore, the content of each trace element was more rationally determined, resulting in less irritation for the user during application. Finally, the product was effective in reducing the white blood cell counts in rats caused by a foreign body implanted in their backs.

[0029] Although the trace element composition of the present application contains similar or identical salts of elements common to Decan products, the product specifications of the trace element composition of the present application are significantly reduced compared to Decan products after preparation into an injection solution. The product specifications are reduced from 40 mL for Decan products to 10 mL for the present application. Taking 50,000 bottles of 40 mL bottles as an example, the optimized 10 mL product of the present application can be made into a batch of 200,000 units, i.e., a larger production lot (four times larger) can be produced for the same volume of product. This significantly reduces the sterilization energy consumption per unit product and the amount of drug transported, significantly reduces production and transportation costs, and is convenient for clinical use. Furthermore, the pH adjuster and pH value of the trace element composition of the present application are different from Decan products, and the trace element composition of the present application has good stability in the pH range of 2.0 to 3.5. Meanwhile, the zinc content of the trace element composition of the present application is slightly lower than that of Decan products. Preliminary safety tests have shown that the composition of the present application has the advantage of fewer side effects.

[0030] In particular, the dosage specifications and pH values ​​of the injectable formulations formed by the trace element composition of the present application differ from those of prior art organic salt-containing products, such as the mainstream 40 mL DECAN products (CN103340895 and CN104971074) commercially available with a pH of 3.5-5.0 (preferably 4.0-4.5) in a 40 mL formulation. Larger formulations require larger packaging volumes, larger manufacturing facilities for the same batch, smaller commercial batches for the same formulation, and larger storage and transportation volumes. In addition, larger formulations affect compatibility with liquids, making clinical application procedures more complicated. For example, the package insert for the aforementioned "DECAN" product states that one 40 mL bottle of the product can be diluted with 250 mL of 0.9% sodium chloride injection, 500 mL of glucose injection, or other suitable parenteral nutrition solutions. Commercially available parenteral nutrition solutions, such as 0.9% sodium chloride injection, glucose injection, or complex amino acid injection, are available in a variety of packaging options, including soft bags, glass bottles, and plastic bottles. Different packaging options for commercially available products have different headspace volumes, which are filled with sterile air or nitrogen. Headspace pressure is often essentially the same as external atmospheric pressure. Some compatible solutions with small headspace volumes can be difficult to continuously add 40 mL of trace elements to. For example, the internal pressure of the compatible solution in a sealed packaging system increases, posing a risk of leakage and inaccurate administration. Alternatively, the headspace volume may be insufficient or difficult to continuously add 40 mL, making clinical applications difficult. Furthermore, an increase in volume can adversely affect the compatible solution itself in terms of osmotic pressure and solution stability, and also prolong the infusion time of the drug solution. The injectable formulation formed by the trace element composition of the present application has a standard of 10 mL, which meets dosage requirements and thus better meets clinical applications. Furthermore, since the pH adjuster of the present application does not contain gluconic acid or gluconic acid lactone, the following problem that arises when gluconic acid or gluconic acid lactone is used as a pH adjuster can be avoided: the pH value of the injection can only be set to a relatively high level, for example, 3.5 to 5.0 (preferably 4.0 to 4.5).At this pH range, the product is not stable after sterilization and may develop a black precipitate over time.

[0031] The research results also showed that even under the conditions of DECAN formulation, after the pH adjuster and stabilizer were changed from gluconic acid lactone to hydrochloric acid, the pH value of the DECAN formulation product increased significantly, and the color of the solution became significantly darker after sterilization. This indicates that the composition of DECAN formulation changes before and after sterilization, and this is probably because hydrochloric acid cannot play the role of chelating elements like gluconic acid lactone, and the valence state of elements changes due to oxidation-reduction reactions between elements, thereby reducing safety.

[0032] When elemental selenium is provided as selenious acid, an inorganic acid and an inorganic base can be simultaneously used as a pH adjuster.

[0033] In some embodiments of the present application, per 10 ml of the trace element composition, the trace elements in the main material are present in the following amounts: 18-21.5 μmol iron, 50-100 μmol zinc, 4.7-6 μmol copper, 1 μmol manganese, 50 μmol fluorine, 0.5-1.0 μmol iodine, 1.0-1.25 μmol selenium, 0.2 μmol molybdenum, and 0.2 μmol chromium.

[0034] In some embodiments of the present application, per 10 ml of the trace element composition, each trace element in the main material is present in the following dosages: 20 μmol iron, 77 μmol zinc, 4.7 μmol or 6 μmol copper, 1 μmol manganese, 50 μmol fluorine, 1 μmol iodine, 1 μmol selenium, 0.2 μmol molybdenum, and 0.2 μmol chromium.

[0035] In some embodiments of the present application, in order to further improve the stability and safety of the trace element composition of the present application, the pH of the trace element composition is preferably 2.6 to 3.2.

[0036] The pH adjuster used in this application is an inorganic acid, and optionally an inorganic base. In order to improve the adjusting efficiency of the pH adjuster, in some embodiments, the inorganic acid is hydrochloric acid or sulfuric acid, preferably hydrochloric acid; and the inorganic base is sodium hydroxide or potassium hydroxide, preferably sodium hydroxide. For example, sodium hydroxide or potassium hydroxide can be added as a pH adjuster when selenium is provided as selenite. For safe use, the pH adjuster can be diluted to a specific concentration in the process, such as 1M hydrochloric acid solution or 1M sodium hydroxide solution.

[0037] The pharmaceutical organic salts of the trace elements iron, zinc, copper, manganese, and chromium of the present application can be various common organic acid salts such as gluconate, fumarate, or amino acid salts. Examples of organic acid salts of iron include ferrous gluconate, ferrous L-aspartate, ferrous DL-aspartate, and ferrous fumarate. Examples of organic acid salts of zinc include zinc gluconate, zinc L-aspartate, and zinc DL-aspartate. Examples of organic acid salts of copper include copper gluconate, copper L-aspartate, and copper DL-aspartate. Examples of organic acid salts of manganese include manganese gluconate, manganese L-aspartate, and manganese DL-aspartate. Examples of organic acid salts of chromium include chromium gluconate, chromium L-aspartate, and chromium DL-aspartate. The pharmaceutical salts of the above trace elements can be anhydrous or hydrated, preferably in a solid form stable at room temperature, such as the relevant pharmaceutical salts listed in domestic and foreign pharmacopeias.

[0038] The pharmaceutical salts of the trace elements of the present invention, fluorine, iodine, molybdenum, and chromium, can be inorganic salts. The pharmaceutical salts of fluorine can be sodium fluoride and potassium fluoride, the pharmaceutical salts of iodine can be sodium iodide and potassium iodide, the pharmaceutical salts of selenium can be sodium selenite and selenious acid, the pharmaceutical salts of molybdenum can be sodium molybdate and ammonium heptamolybdate, and the pharmaceutical salts of chromium can be chromium chloride and chromium sulfate. The pharmaceutical salts of the above trace elements can be anhydrous or hydrated, preferably in the form of a solid stable at room temperature, such as the relevant pharmaceutical salts listed in domestic and foreign pharmacopeias.

[0039] In some embodiments, the main ingredients are ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, potassium or sodium iodide, selenite or sodium selenite, sodium molybdate, and chromium chloride. The ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, potassium or sodium iodide, selenite or sodium selenite, sodium molybdate, and chromium chloride used in this application may be in various crystal water forms or anhydrous forms, and these forms can be converted to the elemental dose of this application without affecting the effectiveness of this application. Selenite or sodium selenite also does not affect the actual effectiveness of this application, and they remain in the same form after adjusting the pH in the solution. Furthermore, testing has shown that when the main ingredients consist of ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, potassium iodide, sodium selenite, sodium molybdate and chromium chloride, the injection preparations formed have better stability and safety.

[0040] In some embodiments of the present application, the components of the above-mentioned trace element composition are mixed to form an injection; preferably, the inner wall of the packaging material of the injection is made of a non-glass packaging material, more preferably, the inner wall of the packaging material is made of a polypropylene material.Using polypropylene as the inner wall of the packaging material can effectively avoid the influence of impurity elements caused by the packaging material.The packaging material can be made entirely of polypropylene material.

[0041] In another exemplary embodiment of the present application, there is provided a method for preparing any of the above trace element compositions, comprising the steps of dispersing salts of each trace element in water for injection, adjusting to a target pH value using a pH adjuster, filtering to obtain a filtrate, adding water to the filtrate to the full volume, potting, and sterilizing.

[0042] The preparation method of the trace element composition of the present application is simple and can be achieved by adopting conventional methods.

[0043] In some embodiments, the above preparation method comprises the steps of: The process involves dissolving and / or suspending the salts of each trace element in water for injection, mixing uniformly, then continuously adding water for injection to complete the dissolution, adjusting the pH value, filtering, adding water for injection to the total volume, potting at 10 mL per bottle, and sterilizing.

[0044] In some embodiments, sterilization is preferably performed using a terminal sterilization process, with an F0 value of ≥8, preferably ≥12. The sterilization temperature and time can be 115°C for 30 minutes or 121°C for 8-20 minutes, preferably 12 minutes at 121°C and 15 minutes at 121°C. The trace element salts are dissolved or suspended in the plastic container, depending on the amount of water added. Even if the salts are prepared as a suspension at an early stage, they can be well dissolved after addition to the mixing tank, allowing for dilution with a large amount of water for injection without affecting the final product volume.

[0045] The dissolution process can involve mixing multiple trace element salts and dissolving them together in water and then mixing them homogeneously; or dissolving multiple trace element salts individually in water and then mixing them homogeneously; or mixing some trace element salts together and then dissolving the remaining trace element salts individually in water and then mixing them homogeneously. Whether multiple trace element salts are dissolved together in water, or some trace element salts are dissolved in water and then dissolving the remaining trace element salts individually in water and then mixing them together, or multiple trace element salts are dissolved individually in water and then mixing them together, the dissolution method does not affect the quality of the final product composition as long as the amount of water added is sufficient (e.g., more than 70% of the specified volume). Here, a sufficient amount of water means at least an amount that allows the trace element salts to dissolve well and achieve a transparent solution. Generally, in terms of "trace amount," even in commercial production, an analytical balance is required to weigh some trace element salts. To ensure the quality stability and consistency of the final composition, the preferred method for accurate weighing and delivery is to dissolve or suspend the trace element salts individually and then mix them together. The amount of water for injection added to dissolve the trace element salts is preferably more than 70%, preferably 80% or more, and more preferably 90% or more of the total amount of the designed composition formulation. To accelerate the dissolution rate, the temperature of the water for injection used for dissolution may be higher than room temperature, for example, 30°C to 70°C.

[0046] In the prior art, commercial products such as "DECAN" and "Addamel" are all packaged with glass materials. Although relevant literature shows that better glass packaging materials, such as borosilicate glass, can better ensure product quality, glass packaging has better air and water vapor isolation performance and higher thermal conductivity than non-glass packaging (PP, PE, etc.). In order to avoid impurity elements brought by packaging materials during the sterilization process, in some embodiments, the packaging material that directly contacts the pharmaceutical product during potting is a non-glass packaging material, such as cyclic olefin copolymer (COC), cyclic olefin polymer (COP), polypropylene (PP) material, preferably polypropylene material.

[0047] The preparation method of the present application does not have any special requirements for the lighting environment, or is carried out in a dark place as specified in the Pharmacopoeia of the People's Republic of China (2020), avoiding direct sunlight, for example. While the illuminance of direct sunlight can reach 60,000 to 100,000 lux, to improve product stability and safety, the trace element composition of the present application is prepared by exposure to illuminance of 500 lux or less, preferably 300 lux or less, even more preferably 100 lux or less, and even more preferably 75 lux or less, thereby improving the stability of the elements in the composition. In production, preparation is preferably carried out in a dark environment using fully automated production equipment. Lighting refers to the lighting environment to which each intermediate product (e.g., the aqueous solution in the dissolution process, the intermediate product after mixing, and the finished aqueous solution) is directly exposed during the preparation process of the trace element composition of the present application. The light source required for preparation may be red or yellow light.

[0048] In a further exemplary embodiment of the present application, there is provided a use of any of the above-mentioned trace element compositions in a medicament for the prevention and treatment of trace element deficiency, so as to meet the requirements of trace element parenteral nutrition, and the medicament has better stability and safety compared to Addaven and Decan products due to the simplified product composition.

[0049] The multi-trace element composition of the present invention can be used to prevent and treat trace element deficiencies, especially in patients who cannot take the composition orally and require parenteral nutrition. The multi-trace element composition of the present invention can meet patients' needs for essential to moderate trace elements, and multiple doses can be used for patients with high demands, such as severe burn patients.

[0050] The advantageous effects of the present application will be further illustrated with reference to the following examples and comparative examples. The following examples are merely illustrative of the technical solutions of the present application and should not be construed as limiting the scope of the claims of the present application.

[0051] The detection methods used in the examples of this application, such as the method for detecting the content of each trace element, may be the same as those in the revised draft of the national standard for "Multi-trace element injection (II)" republished by the Chinese Pharmacopoeia Commission on May 12, 2014 (https: / / www.chp.org.cn / gjyjw / hxyp / 494.jhtml). Because the formulation dosages of this application and Multi-trace element injection (II) are different, the specific amounts of test samples used in this application can be converted to ensure that the contents of the test elements are consistent or similar. If the content falls within the range of the proposed linear regression equation, the element content can be detected using ICP-MS, and iodine can be detected using HPLC. For other parameters, such as pH, transparency and color, appearance, absorbance, visible foreign matter, and insoluble particles, please refer to the test methods and requirements in the Appendix to the Pharmacopoeia of the People's Republic of China. The detection method for the compatible liquid involved in the compatibility stability test can be carried out by referring to the relevant compatible liquid quality standard method. [Example]

[0052] Example 1

[0053] [Table 2]

[0054] According to the above formula, 20L of product was prepared for each design pH value formula for a theoretical amount of 2,000 counts, and the operation was carried out in a dark environment.

[0055] Ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, sodium selenite, sodium molybdate, chromium chloride and potassium iodide were each dissolved or suspended in an appropriate amount of water for injection in a plastic beaker, and the resulting solution or suspension was added to a 20L liquid preparation tank. The plastic beaker was rinsed with water for injection, and the rinse water was added to the liquid preparation tank, followed by stirring until completely dissolved. 1M hydrochloric acid solution was added to adjust the pH to 2.0, 2.2, 2.4, 2.6, 2.9, 3.2, 3.5, 4.0 and 4.3, respectively. Water for injection was added to make up the total volume, and the mixture was mixed uniformly while stirring. The resulting liquid drug was filtered through a filter consisting of two 0.22μm filter cartridges in series, and potted into polypropylene ampoules with a filling volume of 10mL / count, and sterilized at a temperature of 121℃ for 12 minutes.

[0056] 1) Effect of pH value Representative samples are shown in Figure 1. As can be seen from the prototype results in Figure 1, as the pH value increases from left to right, the solution changes from colorless to yellow and gradually becomes darker. Within the proposed pH range, the pH value of the solution remains essentially constant (±0.1) before and after sterilization. The elemental content (iron, zinc, copper, manganese, fluorine, selenium, molybdenum, chromium, and iodine) of samples with pH values ​​of 2.0, 2.2, 2.4, 2.6, 2.9, 3.2, and 3.5 showed no significant changes before and after sterilization, all within 100% ±3% of the theoretical supply. The copper and selenium contents of samples with pH values ​​of 4.0 and 4.3 decreased by 5.7% and 7.8%, respectively, compared to before sterilization.

[0057] 2) Preliminary temperature stability test of the compounded ingredients Samples with pH values ​​of 2.0, 2.2, 2.4, 2.6, 2.9, 3.2, 3.5, 4.0, and 4.3 were collected and stored at 60°C for one month to examine their preliminary stability. The clarity, color, and pH value of the solutions were compared before and after storage at 60°C for one month. The results are shown in Table 3.

[0058] [Table 3]

[0059] The above preliminary stability results show that the color of the solution gradually darkens from colorless to yellow as the pH value increases, and at the same time, the color of the solution gradually darkens as time passes. The color of the product solution with a pH value of 3.5 or higher is obviously darker, and the color of the sample solutions with a pH value of 4.0 and 4.5 is dark yellow, accompanied by a decrease in pH value and black precipitates. This indicates that the product has poor stability and may pose a risk to product quality, especially under conditions of long-term storage in summer and accidental exposure to high temperatures during transportation.

[0060] The detection results for the contents of elements (iron, zinc, copper, manganese, fluorine, selenium, molybdenum, chromium, and iodine) in samples with pH values ​​of 2.0, 2.2, 2.4, 2.6, 2.9, 3.2, and 3.5 showed no obvious changes after one month at 60°C, and all were within 100% ± 3% of the theoretical supply amount. The samples with pH values ​​of 4.0 and 4.3 were characterized by the occurrence of black precipitates, which did not meet the requirements, so the contents of the elements were not further detected.

[0061] 3) Photostability test For example, a sample with a pH of 2.9 was placed under conditions of 500 lux, 300 lux, 100 lux, and 50 lux, respectively, and the absorbance (color) was measured at 325 nm and 420 nm. Absorbance was measured at wavelengths of 325 nm and 420 nm according to UV-visible spectrophotometry (Pharmacopoeia of the People's Republic of China (2020) Part IV General Chapter 0401) using water as a blank. The absorbance results are shown in Table 4.

[0062] [Table 4]

[0063] As can be seen from the photostability results in Table 4, the absorbance did not change significantly within 4 hours at 300 lux, which can meet the requirements for general commercial production. The absorbance was stable at 100 lux, which can meet the process requirements for mass production.

[0064] Example 2 Commercial Formulations and Methods The formulation was the same as in Example 1. According to the formulation, 350 L of product was prepared for a quantity of 35,000 counts, and the operation was carried out in a dark environment.

[0065] Preparation of 1M hydrochloric acid solution: 1000mL of hydrochloric acid was diluted with water for injection to a 1M hydrochloric acid solution, which was mixed uniformly for later use.

[0066] Add the water for injection to the liquid preparation tank and simultaneously start stirring. Dissolve or suspend ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, and sodium fluoride in an appropriate amount of water for injection in a plastic beaker. Add the resulting solution or suspension to the liquid preparation tank and stir to dissolve. Rinse the plastic beaker with an appropriate amount of water for injection and add the rinsed water to the liquid preparation tank. Add sodium selenite, sodium molybdate, chromium chloride, and potassium iodide to the plastic beaker. The plastic beaker is dissolved in an appropriate amount of water for injection in the car, and the resulting solution is added to the liquid preparation tank. The plastic beaker is rinsed with an appropriate amount of water for injection, and the rinsed water is added to the liquid preparation tank. 1M hydrochloric acid solution is added to adjust the pH value to 2.8-3.0, and water for injection is added to make up the total volume. The intermediate solution is sampled for detection (characteristics, pH value, bacterial endotoxin and the content of each trace element). The resulting liquid drug is filtered through a filter with two 0.22μm filter cartridges in series into a buffer tank before the potting machine, and is ready for potting.

[0067] The BFS plastic ampoule potting machine transports polypropylene resin through a vacuum material suction device to the hopper of the BFS extruder, then presses the liquid chemicals in the buffer tank into the liquid separator according to the specified program, starts ampoule production and potting, and fills the product with 10ml / count. The potted product is then transported to the sterilization procedure through a conveyor belt and sterilized at a temperature of 121°C for 12 minutes.

[0068] Vacuum leak detection was performed using a high-voltage electronic leak detector in a water bath sterilization leak detection cabinet according to a specified program, and non-conforming products were rejected. Passing products were subjected to a lamp inspection and packaged.

[0069] The detection results of the element contents (iron, zinc, copper, manganese, fluorine, selenium, molybdenum, chromium and iodine) in the sample showed no obvious changes before and after sterilization, and all were within the range of 100% ± 3% of the theoretical supply amount.In addition, there was no change in the properties and pH value, which all met the requirements.

[0070] Example 3 The formulation was the same as in Example 1, except that the content of the effective element in the copper gluconate was changed from 6 μmol to 4.7 μmol.

[0071] According to the formulation of this example, 10 L of the product was prepared for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment. The preparation method was the same as in Example 2.

[0072] The content of elements (iron, zinc, copper, manganese, fluorine, selenium, molybdenum, chromium and iodine) in the sample was not significantly different before and after sterilization, and all were within the range of 100%±3% of the theoretical supply amount. In addition, there was no change in the properties and pH value, which all met the requirements.

[0073] Example 4 The formulation was the same as in Example 1, except that the content of the active element in copper gluconate was changed from 6 μmol to 4.7 μmol, sodium iodide was used instead of potassium iodide, and selenious acid was used instead of sodium selenite.

[0074] According to the formulation of this example, 10 L of product was prepared for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment. The preparation method was the same as in Example 2, except that the pH adjuster was changed to 1 M hydrochloric acid aqueous solution and 1 M sodium hydroxide aqueous solution.

[0075] The content of elements (iron, zinc, copper, manganese, fluorine, selenium, molybdenum, chromium and iodine) in the sample was not significantly different before and after sterilization, and all were within the range of 100%±3% of the theoretical supply amount. In addition, there was no change in the properties and pH value, which all met the requirements.

[0076] Example 5

[0077] [Table 5]

[0078] According to the above formulation of the main agent, 10 L of product was prepared for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment.

[0079] Ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, sodium selenite, sodium molybdate, chromium chloride, and potassium iodide were each dissolved in an appropriate amount of water for injection in a plastic beaker. The resulting solution was added to a liquid preparation tank. The plastic beaker was rinsed with water for injection, and the rinse water was added to the liquid preparation tank. 1M hydrochloric acid solution was added to adjust the pH to 2.8-3.0. Water for injection was added to the total volume, and the mixture was stirred to homogenize. The resulting liquid formulation was filtered through a filter consisting of two 0.22 μm filter cartridges in series, potted into polypropylene ampoules at a filling volume of 10 mL / count, and sterilized at 121°C for 12 minutes. The product underwent lamp inspection and was then packaged.

[0080] The sterilized product was stored at 60°C for 30 days. The product's quality indicators, including the solution's transparency, color, pH value, and content of each ingredient, showed no obvious changes before and after sterilization or after storing at 60°C for 30 days. The product has good quality stability.

[0081] Example 6

[0082] [Table 6]

[0083] According to the above formulation of the main agent, 10 L of product was prepared for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment.

[0084] Ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, sodium selenite, sodium molybdate, chromium chloride, and potassium iodide were each dissolved in an appropriate amount of water for injection in a plastic beaker. The resulting solution was added to a liquid preparation tank. The plastic beaker was rinsed with water for injection, and the rinse water was added to the liquid preparation tank. 1M hydrochloric acid solution was added to adjust the pH to 2.8-3.0. Water for injection was added to the total volume, and the mixture was stirred to homogenize. The resulting liquid formulation was filtered through a filter consisting of two 0.22 μm filter cartridges in series, potted into polypropylene ampoules at a filling volume of 10 mL / count, and sterilized at 121°C for 12 minutes. The product underwent lamp inspection and was then packaged.

[0085] The sterilized product was stored at 60°C for 30 days. The quality indicators of the product, including the transparency, color, pH value, and content of each ingredient of the solution, showed no significant changes before and after sterilization or after storing at 60°C for 30 days. The product has good quality stability.

[0086] Example 7

[0087] [Table 7]

[0088] According to the above formulation of the main agent, 10 L of product was prepared for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment.

[0089] Ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, selenite, sodium molybdate, chromium chloride, and sodium iodide were each dissolved in an appropriate amount of water for injection in a plastic beaker. The resulting solution was added to a liquid preparation tank. The plastic beaker was rinsed with water for injection, and the rinse water was added to the liquid preparation tank. 1M hydrochloric acid solution or sodium hydroxide solution was added to adjust the pH to 2.8-3.0. Water for injection was added to the total volume, and the mixture was stirred to homogenize. The resulting liquid formulation was filtered through a filter consisting of two 0.22 μm filter cartridges in series, potted into polypropylene ampoules at a filling volume of 10 mL / count, and sterilized at 121°C for 12 minutes. The product underwent lamp inspection and was then packaged.

[0090] The sterilized product was stored at 60°C for 30 days. The product's quality indicators, including the solution's transparency, color, pH value, and content of each ingredient, showed no obvious changes before and after sterilization or after storing at 60°C for 30 days. The product has good quality stability.

[0091] Example 8 The formulation and preparation were the same as in Example 2, except that the packaging materials were low borosilicate glass ampoules and medium borosilicate glass ampoules. 10 L of each product was prepared for a theoretical amount of 1,000 counts.

[0092] The samples of Examples 7 and 8 were subjected to inductively coupled plasma atomic emission spectrometry (ICP) and inductively coupled plasma mass spectrometry (ICP-MS) to detect several impurity elements. The results are shown in Table 8.

[0093] [Table 8]

[0094] When glass ampoules are used, the content of impurity elements is significantly higher, and the content of impurity elements increases significantly after sterilization. However, when polypropylene ampoules are used, the level of impurity elements is extremely low and remains unchanged before and after sterilization. This indicates that the relevant impurity elements should be introduced as raw materials, and that the packaging material does not affect the impurity elements.

[0095] The above-mentioned impurity element tests were conducted on DEACN product (Lot No.: 9901563, Supplier: Laboratoire AGUETTANT). These results show that the B, Si, Ba, and Al contents were 8.92 μg / mL, 0.56 μg / mL, 29.43 μg / mL, and 7.87 μg / mL, respectively. The Al content of each bottle was nearly 315 μg per 40 mL standard (basic daily dose), exceeding the risk management dose for aluminum toxicity for a 50 kg adult (4-5 μg / kg).

[0096] It is clear that the above impurity elements pose safety risks. In most cases, parenteral nutrition supplementation with these preparations continues for a long period of time, and patients receiving parenteral nutrition suffer from underlying diseases and are in poor health. Long-term administration of these medications often leads to liver and kidney dysfunction and deficiency. The USP (United States Pharmacopeia) clearly states that in patients with impaired renal function, including premature infants, cumulative intestinal aluminum absorption exceeding 4-5 μg / kg per day can cause central nervous system and bone toxicity, and lower doses can lead to tissue aluminum overload. Aluminum levels in patients with impaired renal function receiving long-term parenteral nutrition should be monitored regularly to prevent aluminum toxicity.

[0097] As can be seen from the above test results, when polypropylene is used as the packaging material, the trace element composition injection of the present application has a significantly low level of impurity elements, and the impact of the impurity elements contained therein on safety can be ignored.

[0098] Example 9

[0099] [Table 9]

[0100] According to the above formulation of the main agent, 10 L of product was prepared for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment.

[0101] Ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, selenite, sodium molybdate, chromium chloride, and sodium iodide were each dissolved in an appropriate amount of water for injection in a plastic beaker. The resulting solution was added to a liquid preparation tank. The plastic beaker was rinsed with water for injection, and the rinse water was added to the liquid preparation tank. 1M hydrochloric acid solution was added to adjust the pH to 2.8-3.0. Water for injection was added to the total volume, and the mixture was stirred to homogenize. The resulting liquid formulation was filtered through a filter consisting of two 0.22μm filter cartridges in series, potted into polypropylene ampoules at a filling volume of 10mL / count, and sterilized at 121°C for 12 minutes. The product underwent lamp inspection and was then packaged.

[0102] The sterilized product was stored at 60°C for 30 days. The product's quality indicators, including the solution's transparency, color, pH value, and content of each ingredient, showed no obvious changes before and after sterilization or after storing at 60°C for 30 days. The product has good quality stability.

[0103] Comparative Example 1 Based on the formulation of DECAN, the dose of zinc gluconate was increased to 153 μmol.

[0104] [Table 10]

[0105] According to the above formulation, 10 L of product was prepared for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment. The preparation method was the same as in Example 4.

[0106] The detection results of the content of elements (iron, zinc, copper, manganese, fluorine, selenium, molybdenum, chromium, and iodine) in the sample showed no obvious change before and after sterilization, and all were within 100% ± 3% of the theoretical amount supplied. There was no change in the properties and pH value. A comparison between the samples of Comparative Example 1 and Example 9 showed that the solution color of the sample of Comparative Example 1 was darker than that of Example 9 before and after sterilization, indicating that the high content of zinc gluconate may affect the color of the product.

[0107] Comparative Example 2

[0108] [Table 11]

[0109] According to the above formula, 10 L of product was prepared for each design pH value formula for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment.

[0110] Ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, sodium selenite, sodium molybdate, chromium chloride, and potassium iodide were dissolved or suspended in an appropriate amount of water for injection in a plastic beaker, and the resulting solution or suspension was added to a 10-liter liquid preparation tank. The plastic beaker was rinsed with water for injection, and the rinse water was added to the liquid preparation tank, followed by stirring until completely dissolved. A gluconic acid lactone solution was added to adjust the pH to 2.0, 2.2, 2.4, 2.6, 2.9, 3.2, 3.5, 4.0, and 4.3, respectively, and water for injection was added to the total volume, followed by stirring to homogeneity. The subsequent filtration, potting, and stirring processes were the same as in Example 4 (the resulting liquid formulation was filtered through a filter consisting of two 0.22 μm filter cartridges in series, potted into polypropylene ampoules at a filling volume of 10 mL / count, and sterilized at 121°C for 12 minutes). The feasibility was tested under different pH value conditions.

[0111] The test results show that it is difficult to achieve a pH value of less than 3.5 by adjusting the pH using gluconic acid lactone. Adding large amounts of gluconic acid lactone to a pH value of 3.5 barely lowers the pH of the solution. After lowering the pH to 3.3, the pH value of the solution cannot be further lowered. Even if the unit solution exceeds the designed 10 mL specification by adding more gluconic acid lactone, the target lower pH value cannot be achieved. In other words, the above formulation requires a large amount of gluconic acid lactone, and on the other hand, it is difficult to adjust the target lower pH value using gluconic acid lactone, making it difficult to meet the commercial process requirements for products with pH values ​​of 2.0, 2.2, 2.4, 2.6, 2.9, and 3.2. Furthermore, sterilized samples with pH values ​​of 3.5, 4.0, and 4.5 all developed black precipitates after one month at 60°C.

[0112] Similar to the examples in this application, the effect of reducing the dosage of the DECAN unit formulation from 40 mL to 10 mL was also tested. These results are consistent with the above, that is, it is difficult to achieve a sample with a pH value of less than 3.5 by adjusting the pH value with gluconic acid lactone, making it difficult to achieve the commercial process requirements of products with pH values ​​of 2.0, 2.2, 2.4, 2.6, 2.9, and 3.2. Furthermore, sterilized samples with pH values ​​of 3.5, 4.0, and 4.5 all produced black precipitates after being stored at 60°C for less than one month.

[0113] Comparative Example 3 Prepare according to DECAN formula (40 mL / bottle).

[0114] [Table 12]

[0115] The preparation was carried out in a volume 500 times the above formulation (20 L), and the operation was carried out in a dark environment: (1) Sodium fluoride, ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium iodide, cobalt gluconate, ammonium heptamolybdate tetrahydrate, sodium selenite, and chromium chloride were each dissolved in an appropriate amount of water for injection in a plastic beaker, the resulting solution was added to a liquid preparation tank, the plastic beaker was rinsed with water for injection, the rinse water was added to the liquid preparation tank, 1M hydrochloric acid solution was added to adjust the pH value to 4.1 to 4.4, water for injection was added to make up the total volume, and the mixture was mixed uniformly while stirring, and the resulting liquid drug was filtered through a filter consisting of two 0.22 μm filter cartridges in series, potted in 40 mL / bottle, and sterilized at a temperature of 121°C for 12 minutes.

[0116] (2) Sodium fluoride, ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium iodide, cobalt gluconate, ammonium heptamolybdate tetrahydrate, sodium selenite, and chromium chloride were each dissolved in an appropriate amount of water for injection in a plastic beaker, the resulting solution was added to a liquid preparation tank, the plastic beaker was rinsed with water for injection, the rinse water was added to the liquid preparation tank, 20% gluconic acid lactone solution was added to adjust the pH value to 4.1 to 4.4, water for injection was added to make up the total volume, and then the mixture was mixed uniformly while stirring, and the resulting liquid drug was filtered through a filter consisting of two 0.22 μm filter cartridges in series, potted in 40 mL / bottle, and sterilized at a temperature of 121°C for 12 minutes.

[0117] Comparison of the above products before and after sterilization in terms of properties, pH value, etc. showed the following: The pH value of the product solution (1), which was adjusted with hydrochloric acid, was 4.32 before sterilization and 4.60 after sterilization, indicating a clear increase in pH. The color of the solution was a pale yellow, transparent liquid (less yellow-green than standard colorimeter No. 3) before sterilization, and became significantly darker after sterilization (closer to the yellow-green of standard colorimeter No. 5); The pH of product solution (2), which was adjusted using gluconic acid lactone solution, was 4.32 before sterilization and 4.30 after sterilization, with no obvious change in pH. The color of the solution was a pale yellow, transparent liquid (less yellow-green than standard colorimeter No. 3) before and after sterilization, with no obvious change in color. This indicates that if the pH adjuster was changed from gluconic acid lactone to hydrochloric acid under the conditions of DECAN formulation, the solution would become unstable after sterilization, the pH would rise significantly, the color would become significantly darker, and the pH after sterilization would exceed the pH range (4.0-4.5) set by the DECAN standard.

[0118] Comparative Example 4 The composition of the main trace element ingredients for Addaven products is as follows (10mL standard):

[0119] [Table 13]

[0120] According to the above formulation of the main agent, 10 L of product was prepared for a theoretical amount of 1,000 counts, and the operation was carried out in a dark environment.

[0121] Ferric chloride, zinc chloride, copper chloride, manganese chloride, sodium fluoride, sodium selenite, sodium molybdate, chromium chloride and potassium iodide were respectively dissolved or suspended in an appropriate amount of water for injection in a plastic beaker, and the resulting solution or suspension was added to a 10L liquid preparation tank, the plastic beaker was rinsed with water for injection, and the rinse water was added to the liquid preparation tank, and then stirred until completely dissolved, 1M hydrochloric acid solution was added to adjust the pH value to 1.8, 2.0, 2.2, 2.4 and 2.6, and water for injection was added to make up the total volume, and then mixed uniformly while stirring, and the resulting liquid drug was filtered through a filter consisting of two 0.22μm filter cartridges in series, and potted into polypropylene ampoules with a filling volume of 10mL / count, and sterilized at a temperature of 121℃ for 12 minutes.

[0122] Results: The samples with pH values ​​of 2.4 and 2.6 were almost colorless to pale yellow, transparent liquids before sterilization, and yellow, cloudy liquids after sterilization, indicating that the samples without xylitol as a stabilizer did not meet the requirements after terminal sterilization. The samples with pH values ​​of 1.8, 2.0, and 2.2 were all almost colorless, transparent liquids before and after sterilization, indicating that the Addaven product without xylitol could be prepared into a transparent solution sample after further lowering the pH.

[0123] Comparative Example 5 Xylitol stabilizer was added to Addaven products. Similar to the process for the above formulations, ferric chloride, zinc chloride, copper chloride, manganese chloride, sodium fluoride, sodium selenite, sodium molybdate, chromium chloride, and potassium iodide were dissolved or suspended in the appropriate amount of water for injection in a plastic beaker. The resulting solution or suspension was added to a 10 L liquid preparation tank containing dissolved xylitol. The plastic beaker was rinsed with water for injection, and the rinse water was added to the liquid preparation tank. The mixture was stirred until completely dissolved. 1 M hydrochloric acid was added to adjust the pH to 2.4-2.6. Water for injection was added to the total volume, and the mixture was mixed uniformly with stirring. The resulting liquid formulation was filtered through a filter consisting of two 0.22 μm filter cartridges in series, potted into polypropylene ampoules at a fill volume of 10 mL / count, and sterilized at 121°C for 12 minutes. The product was a nearly colorless to pale yellow, transparent liquid before and after sterilization.

[0124] Furthermore, the samples of Comparative Examples 4 and 5 were tested, and the results are shown in Table 14, with the amounts of the ingredients taken as 100%.

[0125] [Table 14]

[0126] The test results show that without xylitol as a stabilizer (i.e., Comparative Example 4), a clear solution can be prepared at a lower pH value, but the iodine content after sterilization is significantly reduced (about 40%), while with xylitol as a stabilizer (i.e., Comparative Example 5), a clear product can be prepared, but again the iodine content is significantly reduced (about 20%). Commercial production of Addaven products requires an excess iodide supply (about 120%).

[0127] Comparing the Examples of the present application with Comparative Examples 4 and 5, the change in iodine content of the formulation of the present application before and after sterilization was within ±3%, indicating that the iodine element can be further stabilized without using a stabilizer in the formulation of the present application.

[0128] Taking the sample of Example 2 as an example, the trace element composition injection of the present application was subjected to accelerated stability test, long-term stability test and compatibility stability test.

[0129] 1) Accelerated and long-term stability tests The polypropylene ampoule inner packaging material and the paper box outer packaging material were subjected to accelerated stability testing (temperature 40°C ± 2°C, RH 25% ± 5%) and long-term stability testing (temperature 25°C ± 2°C, RH 60% ± 5%), respectively. The results are shown in Tables 15 and 16.

[0130] [Table 15]

[0131] [Table 16]

[0132] The results showed that all the indicators in the accelerated stability test showed no obvious changes, meeting the relevant requirements for the drug. Under high temperature and low humidity conditions, a slight increase in the contents occurred due to a slight moisture loss in the packaging, but all requirements were met. Under the long-term stability test conditions (temperature 25°C ± 2°C, RH 60% ± 5%), the water loss rate was less than 0.3% after 12 months, and all quality indicators showed no obvious changes.

[0133] 2) Compatibility and stability testing 0.9% sodium chloride injection, 5% glucose injection, 10% glucose injection, complex amino acid injection (20AA), medium-chain lipid emulsion injection (C 6-24 Compatibility stability tests were conducted with the commercial "All-in-One" and clinical "All-in-One" formulations. Compatibility tests with sodium chloride injection, glucose injection, complex amino acid injection, and medium-chain lipid emulsion injection were conducted as follows: 1 count (10 mL) of each injection was added to 500 mL of compatible solution. If the total volume was less than 500 mL, the appropriate proportions were calculated. The "All-in-One" compatible solution was prepared by adding 1 count (10 mL) of the product, as well as 1 count each of water-soluble vitamins and fat-soluble vitamins. The lipid emulsion injection and "All-in-One" compatibility systems are complex, and the primary focus was on determining their properties, pH, osmolality, particle size and particle size distribution, and PFAT5 index.

[0134] The content at time 0 is taken as 100%. Regarding compatibility with glucose injection, the Pharmacopoeia of the People's Republic of China (2020) is referred to for 5-hydroxymethylfurfural in glucose injection, and the "Study on the Limit Test for 5-hydroxymethylfurfural in Vitamin C Glucose Injection" is referred to for the limit and method of 5-hydroxymethylfurfural. The detection limit by high-performance liquid chromatography must be 0.02% or less. The content of some amino acids was detected using an amino acid analyzer. The test results are shown in Tables 17-22.

[0135] [Table 17]

[0136] [Table 18]

[0137] [Table 19]

[0138] [Table 20]

[0139] [Table 21]

[0140] [Table 22]

[0141] As can be seen from the above compatibility stability test, the composition injection of the present application has good compatibility stability with common compatible solutions.

[0142] 3) Safety testing Samples from Example 2 (test sample group) and Addaven product (control sample group, lot number: 12NHB19, supplier: Fresenius Kabi Limited) were collected and compared in safety tests including a systemic active allergic reaction test in guinea pigs, a hemolysis test in New Zealand male rabbits, and a vascular irritation test in New Zealand male rabbits.

[0143] Systemic active allergic reaction test in guinea pigs: Hartley guinea pigs, SPF grade, supplier: Beijing Vital River Laboratory Animal Technology Co., Ltd., animal production permit number: SCXK(JING)2016-0011, test animal certification number: No. 110011201110221326, issuer: Beijing Municipal Science and Technology Commission, age at start of administration: 5-6 weeks.

[0144] Guinea pigs were sensitized with the sample from Example 2 and Addaven by intraperitoneal injection every other day for three consecutive days. For the allergic reaction test, 0.9% sodium chloride injection (lot number: 6B20060706, Shandong Qidu Pharmaceutical Co., Ltd.) was selected as the compatible solution. The low-dose test sample group was administered the clinically intended concentration of the test sample solution, while the high-dose test sample group was administered a test sample solution with double the intended concentration. The dose was 0.5 mL per animal in each group. On the 14th and 21st days after the final sensitization, the product was intravenously injected at twice the sensitizing dose for stimulation.

[0145] Low-dose test sample group: 4 mL of the sample of Example 2 was added to 40 mL of 0.9% sodium chloride injection in a clean bench and shaken well to obtain the administration preparation for the low-dose test sample group, which was then sealed and stored at room temperature in a dark place. High-dose test sample group: 5 mL of the sample of Example 2 was added to 25 mL of 0.9% sodium chloride injection in a clean bench and shaken well to obtain the administration preparation for the high-dose test sample group, which was then stored in a sealed container at room temperature in a dark place. Control sample group: The dosage preparation was prepared and stored according to the same method as the high dose test sample group using Addaven product. Negative control group: 0.9% sodium chloride injection; Positive control group: An appropriate amount of bovine serum albumin (Lot No. 128K054, Beijing Solebao Science and Technology Co., Ltd.) was weighed and prepared into a bovine serum albumin solution at a concentration of 40 mg / mL using 0.9% sodium chloride injection.

[0146] The guinea pig systemic active allergic reaction test protocol is shown in Table 23.

[0147] [Table 23]

[0148] Test Results: No allergic reactions were observed in the animals in each dose group of the sample from Example 2 and Addaven, consistent with the results of the negative control group (0.9% sodium chloride injection). During the test, no abnormal reactions were observed in the clinical observations of the animals, and the weight gain of the animals in each group was normal. After stimulation on the 14th day from the last sensitization, animals in the positive control group showed symptoms of allergic reaction, including piloerection, nasal scratching, coughing, urination, dyspnea, wheezing, unsteady gait, convulsions, periodic breathing, and death, with the allergic reaction rated as strongly positive to very strongly positive. After stimulation on the 21st day from the last sensitization, animals in the positive control group showed symptoms of allergic reaction, including piloerection, nasal scratching, sneezing, coughing, urination, defecation, dyspnea, wheezing, unsteady gait, convulsions, periodic breathing, and death, with the allergic reaction rated as strongly positive to very strongly positive.

[0149] A systemic active allergic reaction test in guinea pigs showed that both the injection of the present application and Addaven caused negative allergic reactions.

[0150] 4) In vitro hemolysis test using rabbit red blood cells 2% Rabbit Erythrocyte Suspension: 8 mL of blood was collected from the central ear artery of a healthy male New Zealand rabbit, placed in an Erlenmeyer flask containing glass beads, and shaken for approximately 10 minutes. The blood was then stirred with a glass rod to remove fibrinogen and obtain defibrinated blood. Erythrocytes were aspirated using a pipette, and then 10 volumes of sodium chloride injection were added. The mixture was shaken thoroughly and centrifuged at 1500 rpm for 10 minutes. After removing the supernatant, the precipitated red blood cells were washed five times in the same manner, four times with sodium chloride injection, until the supernatant was no longer red. 0.6 mL of the resulting red blood cells was collected and resuspended in sodium chloride injection at a 2% volume ratio for further use.

[0151] Test animal producer: Suzhou Genesc Biotechnology Co., Ltd., Test animal quality certification number: 20201103271, Test animal production permit number: SCXK(SU)2020-0002, Issued by: Jiangsu Provincial Science and Technology Bureau.

[0152] Test sample solution: 9 mL of the sample of Example 2 was added to 90 mL of 0.9% sodium chloride injection in a clean bench and shaken well to obtain a test sample solution, which was then sealed and stored at room temperature in a dark place. Control sample solution: 9 mL of Addaven product was added to 90 mL of 0.9% sodium chloride injection in a clean bench and shaken well to obtain a control sample solution, which was then sealed and stored at room temperature in a dark place. Negative control solution: 0.9% sodium chloride injection. Positive control solution: sterile water for injection (Lot No. 2A20032101, Shandong Qidu Pharmaceutical Co., Ltd.).

[0153] The clean test tubes were numbered as follows: test tubes 1-5 and 8-12 were test sample / control sample solution test tubes, respectively; test tubes 6 and 13 were negative control test tubes (0.9% sodium chloride injection); and test tubes 7 and 14 were positive control test tubes (sterile water for injection). The solutions were added according to the table below, gently shaken, and immediately incubated in an incubator at 37.0°C. Observations were made once every 15 minutes (0 hours before incubation) after adding the solutions to the incubator, and once every hour thereafter for a total of 3 hours, and the results of each observation were recorded.

[0154] [Table 24]

[0155] Test Results: After incubating the 0.9% sodium chloride injection test tubes (6 and 13) at 37.0°C for 3 hours, a large number of red blood cells settled to the bottom of the tube, and the supernatant was colorless and transparent. After vigorously shaking, the settled red blood cells redispersed in each test tube without hemolysis or coagulation. After incubating the sterile water for injection test tubes (7 and 14) at 37.0°C for 3 hours, the solution turned clear red, with no red blood cells remaining at the bottom of the test tube, indicating complete hemolysis. When test sample / control sample solution test tubes (1-5 and 8-12) were incubated at 37.0°C, the erythrocyte sedimentation rate of each test sample / control sample solution test tube became faster than that of the negative control test tube starting from the 30th minute. After 3 hours of incubation, the supernatant of each test sample / control sample solution test tube was colorless and transparent without hemolysis. After vigorously shaking, the settled red blood cells were not uniformly dispersed, and both showed slight coagulation.

[0156] The results of the hemolysis test showed that the injection containing trace elements of the present invention is consistent with that of Addaven product, and both have no hemolytic effect on rabbit red blood cells in vitro and only a slight coagulation effect.

[0157] Here, Figure 2 is a photograph of test tubes Nos. 1 to 7 (the test tubes are numbered 1, 2, 3, 4, 5, 6, and 7 from left to right) that were shaken well after sample addition; Figure 3 is a photograph of test tubes Nos. 1 to 7 3 hours after sample addition; and Figure 4 is a photograph of test tubes Nos. 1 to 7 that were shaken well 3 hours after sample addition.

[0158] 4) Vascular irritation test Ten New Zealand rabbits, 3-4 months old, clean grade, half male and half female. Supplier: Qingdao Kangda Biotechnology Co., Ltd., Permit Number: SCXK(LU)20210003, Test Animal Quality Certification Number: 370823211100104051.

[0159] The preparation methods of the test sample solution, control sample solution and negative control solution are the same as those under the in vitro hemolysis test section.

[0160] In this study, 10 mL of the sample of Example 2 plus 100 mL of 0.9% sodium chloride injection (test sample group) and 10 mL of Addaven product plus 100 mL of 0.9% sodium chloride injection (control sample group) were administered at 5.2 mL / kg, respectively, while 0.9% sodium chloride injection was used as a negative control, and the administration volume was the same as that of the test sample. The administration schedule and grouping are shown in Table 25.

[0161] [Table 25]

[0162] Administration route: marginal ear vein infusion; administration frequency and duration: once daily for 7 consecutive days.

[0163] Recovery period: Animals were observed for 72 hours after the final administration, some animals were subjected to histopathological examination at necropsy, and observation continued for 14 days during the recovery period.

[0164] Administration method: Each group of New Zealand rabbits was administered using a matched left-right self-control method. In this study, a microinjection pump was used for intravenous administration. Test sample solution or control sample solution was administered to the left ear, and 0.9% sodium chloride injection as a control was administered to the right ear using the same method.

[0165] Detailed clinical observations: Observations once daily. Observations include, but are not limited to, physical signs, general activity, mental status, respiratory status, fecal characteristics, and mortality.

[0166] Weight measurement time and animals: The weights of all animals on the day of the first administration and the animals subjected to necropsy on the days of sacrifice and sampling were measured once each.

[0167] Local irritation at the injection site was visually observed before administration on administration days and once daily on non-administration days. 72 hours after the last administration and after 14 days of continuous recovery, the corresponding animals were euthanized under anesthesia (numbers are shown in the table above, T corresponds to the test sample group, and R corresponds to the control sample group). Both rabbit ears were cut at the base of the ear, and the specimens were fixed in 10% neutral formalin solution. After collection, they were dehydrated, embedded in paraffin, sectioned, and stained with HE, followed by histopathological examination.

[0168] Results: During the test, the general clinical signs of the New Zealand rabbits in each group were good, and no obvious abnormalities were observed. The weight of the surviving New Zealand rabbits in each group fluctuated within the normal range, and no obvious abnormal changes were observed.

[0169] Furthermore, during the period from day 5 to day 10, all animals in the control sample group showed intravascular congestion at the administration site on the administration side, whereas no obvious abnormalities were visually observed at the administration site in either the test sample group of the present application or the control side of the above animals.

[0170] Seventy-two hours after the last administration, the blood vessels at the injection site and proximal non-injection site on the administration side of all animals in the control sample group were reddish-purple. Microscopic observation showed intravascular congestion, vasodilation, and inflammatory cell infiltration into the perivascular tissue in the proximal non-injection site of the ear vein on the administration side of all animals in the control sample group, suggesting that marginal ear vein infusion of the control sample can cause significant inflammation in the ear vein and surrounding tissue of New Zealand rabbits. No abnormalities were observed in the blood vessels at the injection site on the control side of the test sample group or other animals.

[0171] At the end of the recovery period, no abnormalities were observed in the distal non-injected site, the injected site, or the proximal non-injected site on the treated and control sides in all animals in the test sample and control sample groups. The vascular pattern was clear, with no swelling or congestion, and no pathological changes such as hemorrhage or edema were observed in the perivascular tissue.

[0172] In the vascular irritation test, 72 hours after the final administration, the control sample group showed significant irritation in the ear veins and surrounding tissues of New Zealand rabbits when administered intravenously through the marginal ear vein once a day for 7 consecutive days. In contrast, there was no obvious irritation in the test sample group of the present application. At the end of the recovery period, no vascular irritation reaction was observed in the test sample group or the control sample group. Meanwhile, this indicates that the rabbits were able to recover from the irritation of the control sample group after the recovery period.

[0173] 5 to 8 show typical histopathological examination results of animals administered with the control sample and the test sample injection of the claimed composition to the administration and control sides. At the end of the recovery period, no abnormalities were observed in the control group, and no diagrams relating to this are shown in the figures.

[0174] The sample of Comparative Example 1 (lot number: 20220501, supplier: Beijing Zang WeiXinkang Pharmaceutical R & D Co., Ltd.) was subjected to a vascular irritation test in the same manner as above.

[0175] Five New Zealand rabbits, 3-4 months old, clean grade, two females and three males. Test sample group: 10 mL of the sample of Comparative Example 1 was added to 100 mL of 0.9% sodium chloride injection in a clean bench and shaken well to obtain a test sample solution, which was then stored in a sealed container at room temperature in a dark place. Negative control group: 0.9% sodium chloride injection.

[0176] The test samples and negative control were each administered at 5.2 mL / kg. The administration schedule and groupings are shown in Table 26.

[0177] [Table 26]

[0178] Route of administration: Marginal ear vein infusion. Frequency and duration of administration: Once daily for 7 consecutive days.

[0179] Recovery period: Animals were observed for 72 hours after the final administration, some animals were subjected to histopathological examination at necropsy, and observation continued for 14 days during the recovery period.

[0180] Administration method: Each group of New Zealand rabbits was administered using a matched left-right self-control method. In this study, a microinjection pump was used for intravenous administration. The test sample solution was administered to the left ear, and 0.9% sodium chloride injection as a negative control was administered to the right ear using the same method.

[0181] Detailed clinical observations: Observations once daily. Observations include, but are not limited to, physical signs, general activity, mental status, respiratory status, fecal characteristics, and mortality.

[0182] Weight measurement time and animals: The weights of all animals on the day of the first administration and the animals subjected to necropsy on the days of sacrifice and sampling were measured once each.

[0183] Local irritation at the injection site was visually observed before administration on administration days and once daily on non-administration days. 72 hours after the last administration and after 14 days of continuous recovery, the corresponding animals were euthanized under anesthesia (numbers are shown in the table above, with T corresponding to the test sample group). Both rabbit ears were cut at the base of the ear, and the specimens were fixed in 10% neutral formalin solution. After collection, they were dehydrated, embedded in paraffin, sectioned, HE stained, and subjected to histopathological examination.

[0184] Results: During the study, one male New Zealand rabbit died three days after the final dose and was observed to have mild convulsions in the limbs, soiling around the anus, and a prone position. This animal produced small to medium amounts of loose / soft brown feces from three days after the first dose until two days after the final dose. After death, its body weight decreased by 36% compared to before administration, and no obvious abnormal changes were observed upon gross dissection, nor were any lethal lesions observed under microscope. In addition to the dead animal, another male animal produced a small amount of loose feces three days after the first dose, a large amount of loose brown feces from two to four days after the final dose, and then a small amount of loose brown feces. One female animal produced a small amount of loose brown feces from four days after the first dose until three days after the final dose, but the other two animals showed no obvious abnormal reactions through general clinical observation.

[0185] During the entire administration period and recovery period, no obvious irritation-related abnormalities were visually observed at the administration site on the test sample side and the control side of the animals, including the animals that died. No abnormalities were observed at the distal non-injection site, the injection site, or the proximal non-injection site on the administration side and the control side of the animals, and no pathological changes such as hemorrhage and edema were observed in the perivascular tissue.

[0186] No significant abnormalities were observed in vascular irritability in animals administered the sample of Comparative Example 1, which contained a significantly increased zinc dose, but the formulation of Comparative Example 1, which contained a high zinc dose, caused animal deaths and gastrointestinal side effects in some animals.

[0187] Comparative Example 6 The dose of zinc gluconate was increased to 153 μmol.

[0188] [Table 27]

[0189] According to the above formula, 350 L of product was prepared for a theoretical amount of 35,000 counts, and the operation was carried out in a dark environment.

[0190] Water for injection is added to the liquid preparation tank, stirring is started, and while maintaining the stirring state, ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, and sodium fluoride are dissolved or suspended in an appropriate amount of water for injection in a plastic beaker, and the resulting solution or suspension is added to the liquid preparation tank and stirred to dissolve. The plastic beaker is rinsed with an appropriate amount of water for injection, and the rinse water is added to the liquid preparation tank. Sodium selenite, sodium molybdate, chromium chloride, and potassium iodide are dissolved in an appropriate amount of water for injection in a plastic beaker, and the resulting solution is added to the liquid preparation tank. The plastic beaker is rinsed with water for injection, and the rinse water is added to the liquid preparation tank. 1M hydrochloric acid solution is added to adjust the pH to 2.8-3.0, and water for injection is added to make up the total volume. The resulting liquid drug is filtered through a filter consisting of two 0.22 μm filter cartridges in series into a buffer tank before the potting machine, and is ready for potting.

[0191] The BFS plastic ampoule potting machine transports polypropylene resin through a vacuum material suction device to the hopper of the BFS extruder, then presses the liquid chemicals in the buffer tank into the liquid separator according to the specified program, starts ampoule production and potting, and fills the product with 10ml / count. The potted product is then transported to the sterilization procedure through a conveyor belt and sterilized at a temperature of 121°C for 12 minutes.

[0192] The blending process of Example 2 and the sample of Comparative Example 6 were analyzed. The element content was expressed as 100% based on the amount of the blend. The contents of formic acid and oxalic acid, impurities resulting from the decomposition of gluconic acid, were detected by ion chromatography, and the ratio of these to the theoretical amount of gluconic acid in the blend was calculated. The results are shown in Table 28.

[0193] [Table 28]

[0194] The results show that there was no significant change in pH or content before and after sterilization for the formulation of Example 2 and the formulation of Comparative Example 6, but the clarity and color of the solution after sterilization were darker than before sterilization. The formulation of Comparative Example 6 showed a greater change between before and after sterilization than the formulation of Example 2. It can be seen that the content of impurities due to degradation increased after sterilization compared to before sterilization for both the formulation of Example 2 and the formulation of Comparative Example 6, but the increase was more significant for the formulation of Comparative Example 6 than for the formulation of Example 2. Assuming that the gluconic acid content of the formulation of Example 2 is 40 mg and the gluconic acid content of the formulation of Comparative Example 6 is 69 mg, the formic acid and oxalic acid contents in each finished product using the formulation of Example 2 after sterilization were 0.2 mg and 0.24 mg, respectively, while the formic acid and oxalic acid contents in each finished product using the formulation of Comparative Example 6 after sterilization were 0.9 mg and 1 mg, respectively, which are significantly higher than those of Example 2. This indicates that the compositions of the present application have a low impurity content, further reducing the safety risk of impurities that may be present.

[0195] Plasma elemental concentrations, white blood cell count (WBC) and repeated dose irritation tests The plasma concentrations of trace elements, white blood cell count (WBC) and administration irritability were evaluated by observing xenotransplant malnutrition model rats after continuous intravenous injection of three formulations of trace element injections.

[0196] The study was conducted using 40 male SD rats. Grade: SPF grade. Breeder: Beijing Vital River Laboratory Animal Technology Co., Ltd.; Production permit number and issuer: SCXK(JING)2021-0011 (Beijing Science and Technology Commission).

[0197] The animals were fed a diet (containing no trace elements) provided by Tianjin Keao Xieli Feed Co., Ltd. for over five days, and then subcutaneously implanted with cotton balls and filter paper on their backs (eight animals served as normal controls). Three days after surgery, the animals were randomly assigned to a model control group, Combination 1 group (Addaven product), Combination 2 group (Comparative Example 6), and Combination 3 group (Example 2), with eight animals in each group. The test groups and dosing schedules are listed in Table 29.

[0198] [Table 29]

[0199] Starting on the fourth day after surgery, the animals were administered the drug via the tail vein once daily for seven consecutive days at a dose of 10 mL / kg. (The recommended adult clinical dose for Combination Drugs 1 to 3 is 10 mL per day, and for a 60 kg human, the body surface area is 1.62686 m.) 2 For a 200g rat, the body surface area is 0.03036m 2 Therefore, the equivalent clinical dose in rats is approximately 0.93 mL / kg / day. In this study, the dose was 2.5 mL / kg, and a 4-fold diluted injection solution was used, with an administration volume of 10 mL / kg, which is approximately 2.7 times the equivalent clinical dose in rats. The normal control group and model control group were administered 0.9% sodium chloride injection. Each combination group was administered the corresponding injection number.

[0200] PK blood sampling: Approximately 0.2 mL of venous blood was collected from the jugular sinus of rats on the morning of day 1 before surgery, the morning of day 1 after surgery, 0 min before the first dose, 2 min, 15 min, 1 hour, and 4 hours after the first dose, 15 min after the first dose on day 4, 15 min after the first dose on day 7, and the morning of day 10. The whole blood was placed in an EDTA anticoagulant tube and centrifuged at 6000 rpm for 10 min. Plasma was temporarily stored in a refrigerator at -20°C during blood collection. After blood collection, the plasma was transferred to -70°C and frozen for storage, then transported to a central laboratory. The copper, manganese, zinc, and selenium contents in the plasma were detected using an Agilent ICP-MS method.

[0201] The plasma concentrations of copper, manganese, zinc, and selenium detected by ICP-MS showed that the plasma copper concentration increased slightly and the plasma selenium concentration decreased after surgery, while the manganese and zinc concentrations showed no significant changes before and after surgery. After administration, the plasma concentrations increased significantly, reaching their peaks 2 minutes later. The zinc concentrations in groups 2 and 3 basically decreased to the level of the normal control group after 4 hours, and the zinc concentration in group 1 was significantly higher than that in groups 2 and 3. On the 10th day after 7 consecutive days of administration, the plasma zinc concentrations in groups 1, 2, and 3 basically remained normal and were the same as those in the normal control group. The plasma manganese concentrations in groups 1, 2, and 3 were slightly higher than those in the normal control group. The plasma copper and selenium concentrations in groups 1, 2, and 3 remained high and significantly higher than those in the normal control and model control groups. The plasma concentration results for copper, manganese, zinc and selenium are shown in Figures 9, 10, 11 and 12.

[0202] White blood cell count (WBC): Blood samples were collected from the jugular sinus of rats on day 0 before administration and day 4 after administration, anticoagulated with EDTA, and detected using the XN-1000V (BI) fully automated modular animal blood and body fluid analyzer (flow cytometry + semiconductor laser + nucleic acid fluorescent staining method). The results are shown in Figures 13 and 14.

[0203] The results showed a significant increase in postoperative white blood cell counts in both the model and treatment groups. Four days after administration, white blood cell counts in each treatment group were slightly decreased compared to the model group, with Combination Preparations 2 and 3 being comparable to or superior to Combination Preparation 1. Figure 15 shows typical irritation profiles after four consecutive days of tail vein administration in the normal control, model control, Combination Preparation 1, Combination Preparation 2, and Combination Preparation 3 groups. Results of repeated tail vein administration showed no irritation in the normal control and model control groups, but both Combination Preparations 1 and 2 caused relatively strong irritation following tail vein administration. Combination Preparation 1 was more irritating than Combination Preparation 2, while Combination Preparation 3 was almost irritating, demonstrating that Combination Preparations 1 and 2 were significantly more irritating than each other.

[0204] The above results indicate that Combination 1, Combination 2, and Combination 3 showed consistent trends in the plasma concentrations of copper, manganese, zinc, and selenium in rats treated with xenotransplantation malnutrition. All three significantly increased the plasma concentrations of copper, manganese, zinc, and selenium. Because the zinc concentration in Combination 2 was significantly higher than that in Combination 1 and Combination 3, the zinc plasma concentrations in rats treated with Combination 2 were also significantly higher than those in rats treated with Combination 1 and Combination 3. However, both returned to normal control levels by day 10. Combination 2 and Combination 3 were comparable in terms of reducing white blood cell counts in the model animals; that is, there was no significant difference between Combination 2 and Combination 3 in terms of the supplementation effects of various elements. However, Combination 3 was superior to Combination 1 and Combination 2 in terms of irritation caused by repeated intravenous administration. Multiple doses are necessary for patients with strict clinical requirements for trace elements, such as those with severe burns. Formulation 3, which has the advantage of less irritation upon repeated administration, may better meet the safety requirements of clinical application.

[0205] The above examples merely represent some embodiments of the present application, and although the descriptions thereof are relatively specific and detailed, they should not be understood as limiting the scope of the present application. It should be noted that various changes and modifications can be made by those skilled in the art without departing from the spirit of the present application, and all of them are within the scope of protection of the present application. Therefore, the scope of protection of the present application shall be governed by the appended claims.

Claims

1. A trace element composition, comprising a main material and auxiliary materials, the main material being a pharmaceutical organic acid salt of iron, a pharmaceutical organic acid salt of zinc, a pharmaceutical organic acid salt of copper, a pharmaceutical organic acid salt of manganese, a pharmaceutical salt of fluorine, a pharmaceutical salt of iodine, a pharmaceutical salt of selenium, a pharmaceutical salt of molybdenum, a pharmaceutical organic acid salt of chromium, or a pharmaceutical inorganic salt; the auxiliary materials being water for injection and a pH adjuster, the pH adjuster being an inorganic acid and any inorganic base; The pH value of the main composition is 2.0 to 3.5, and the trace elements in the main material are contained in the following amounts per 10 mL of the trace element composition: 17.8 to 21.5 μmol of iron, 40 to 100 μmol of zinc, 4.7 to 9.6 μmol of copper, 1 μmol of manganese, 25 to 60 μmol of fluorine, 0.5 to 1.2 μmol of iodine, 0.75 to 1.27 μmol of selenium, 0.2 to 0.26 μmol of molybdenum, and 0.2 to 0.4 μmol of chromium.

2. 2. The trace element composition of claim 1, wherein per 10 ml of the trace element composition, the trace elements in the main material are present in the following amounts: 18-21.5 μmol of iron, 50-100 μmol of zinc, 4.7-6 μmol of copper, 1 μmol of manganese, 50 μmol of fluorine, 0.5-1.0 μmol of iodine, 1.0-1.25 μmol of selenium, 0.2 μmol of molybdenum, and 0.2 μmol of chromium.

3. 2. The trace element composition of claim 1, wherein per 10 ml of the trace element composition, the trace elements in the main material are in the following amounts: 20 μmol iron, 77 μmol zinc, 4.7 μmol or 6 μmol copper, 1 μmol manganese, 50 μmol fluorine, 1 μmol iodine, 1 μmol selenium, 0.2 μmol molybdenum and 0.2 μmol chromium.

4. The trace element composition according to any one of claims 1 to 3, wherein the pH value of the composition is between 2.6 and 3.

2.

5. 5. The trace element composition according to any one of claims 1 to 4, wherein the inorganic acid is hydrochloric acid or sulfuric acid, preferably hydrochloric acid; and the inorganic base is sodium hydroxide or potassium hydroxide, preferably sodium hydroxide.

6. The organic acid salt of elemental iron is selected from any one of ferrous gluconate, ferrous L-aspartate, ferrous DL-aspartate, and ferrous fumarate; the organic acid salt of elemental zinc is selected from any one of zinc gluconate, zinc L-aspartate, and zinc DL-aspartate; the organic acid salt of elemental copper is selected from any one of copper gluconate, copper L-aspartate, and copper DL-aspartate; and the organic acid salt of elemental manganese is selected from any one of manganese gluconate, manganese L-aspartate, and manganese DL-aspartate. the pharmaceutical salt of elemental fluorine is sodium fluoride or potassium fluoride, the pharmaceutical salt of elemental iodine is sodium iodide or potassium iodide, the pharmaceutical salt of elemental selenium is sodium selenite or selenious acid, the pharmaceutical salt of elemental molybdenum is sodium molybdate or ammonium heptamolybdate, the pharmaceutical inorganic salt of elemental chromium is chromium chloride or chromium sulfate, and / or the organic acid salt of elemental chromium is selected from any one of chromium gluconate, chromium L-aspartate, and chromium DL-aspartate; Preferably, the main ingredients are ferrous gluconate, zinc gluconate, copper gluconate, manganese gluconate, sodium fluoride, potassium or sodium iodide, sodium selenite or sodium selenite, sodium molybdate and chromium chloride; The trace element composition according to any one of claims 1 to 5.

7. The trace element composition according to any one of claims 1 to 6, wherein the trace element composition is an injection; preferably, the inner wall of the packaging material of the injection is made of a non-glass packaging material; more preferably, the inner wall of the packaging material is made of a polypropylene material.

8. The method comprises the steps of dispersing the salts of the respective trace elements in water for injection, adjusting the pH to a target value using a pH adjuster, filtering to obtain a filtrate, adding water to the filtrate to make up the total volume, potting, and sterilizing, and the sterilization is preferably carried out using a terminal sterilization step, and the F of the terminal sterilization step ０ The method for preparing a trace element composition according to any one of claims 1 to 7, wherein the value is 8 or more, preferably 12 or more.

9. the packaging material that comes into direct contact with the medicine during potting is a non-glass packaging material, preferably a polypropylene material; Preferably, the trace element composition is prepared under conditions of exposure to an illumination intensity of 500 lux or less, preferably an illumination intensity of 300 lux or less, more preferably an illumination intensity of 100 lux or less. The preparation method according to claim 8.

10. Use of the trace element composition according to any one of claims 1 to 7 in a medicament for the prevention and treatment of trace element deficiency.

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