Sodium danshensu injection and industrial production method therefor

Abstract

A sodium danshensu injection and an industrial production method therefor. The injection comprises sodium danshensu and a stabilizer. The concentration of the stabilizer is 1 / 2 to 1 / 50 of the concentration of sodium danshensu, and the injection has a pH of 1-6. The stabilizer is disodium edetate or sodium calcium edetate. Also provided is an industrial production method for the sodium danshensu injection. The sodium danshensu injection has good stability, good clinical safety, a simple preparation process, and suitable costs, and is thus suitable for industrial large-scale production. Clinically, the sodium danshensu injection is administered by means of direct intravenous injection or, after being prepared as an infusion, intravenous drip infusion, which is convenient and quick.

Description

A sodium tanshinone injection and its industrial production method Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a sodium tanshinone injection and its industrial production method. Background Technology

[0002] Danshensu, chemically named [D-(+)-β-(3,4-dihydroxyphenyl)lactic acid], is usually stored as a sodium salt due to its unstable structure. Sodium tanshensu has the molecular formula C9H9O5Na, a molecular weight of 220.15, and the structural formula is as follows:

[0003] Tanshinone is one of the main active components of Salvia miltiorrhiza, and its pharmacological effects have been extensively studied by scholars both domestically and internationally. It has been found that tanshinone possesses free radical scavenging, antioxidant, anti-inflammatory, anti-fibrotic, anti-lipid peroxidation, anti-apoptotic, anti-autophagy, and anti-calcium properties. 2+ Danshen sodium exhibits a variety of pharmacological activities, including overload, inhibition of platelet aggregation, inhibition of thrombus formation, prevention of atherosclerosis, protection of vascular endothelial cells, protection against myocardial ischemia and hypoxia, dilation of coronary arteries, improvement of endothelial progenitor cell function, improvement of microcirculation, improvement of mitochondrial function, and immunomodulation of lipid metabolism. It has protective effects on the cardiovascular system, liver, kidneys, and lungs, and is one of the main components of Danshen for the treatment of cardiovascular and cerebrovascular diseases (Zhang Jinli, et al. Eur J Pharmacol. 2019, 864:172710; Bao Xiaoyi, et al. Front Pharmacol. 2018, 9:1445). Therefore, developing tanshinone sodium into a clinically applicable drug for the treatment of coronary microvascular diseases and other cardiovascular and cerebrovascular diseases is a challenging problem that pharmaceutical researchers need to solve.

[0004] Tanshinone sodium contains catechol and lactic acid structures in its molecular structure, exhibiting strong antioxidant activity but also being easily oxidized. Catechol compounds are typical metal chelating agents, capable of forming stable metal chelates with metal ions. Tanshinone sodium is extremely unstable in aqueous solution to heat and oxidants, readily forming complexes with metal ions in solution, and is further degraded during high-temperature sterilization. This leads to tanshinone sodium injection solutions being prone to deterioration during storage, exhibiting quality problems such as significant pH decreases, noticeable color changes, precipitation of unidentified substances, and marked reduction in biological activity. When tanshinone sodium is mixed with commonly used injectable solutions (such as glucose injection, sterile water for injection, and 0.9% NaCl solution), precipitation often occurs over time, severely affecting product stability and clinical use. Therefore, the stability of tanshinone sodium preparations has become a key technology in drug development.

[0005] Chinese patent CN100444836C discloses an injectable pharmaceutical formulation containing sodium tanshinone. However, this formulation includes excipients far exceeding the safe dosage range for sodium tanshinone, meaning the excipients directly enter the bloodstream, posing a relatively high risk for clinical use. Furthermore, this patent lacks in-depth research on the properties of the injectable formulation. Chinese patent CN102614166B discloses an injectable solution prepared from hydroxysaffron yellow pigment A and sodium tanshinone, simply reconstituted with water for injection in a certain proportion. This patented formulation is easily oxidized and unstable. To address this stability issue, most current research focuses on developing sodium tanshinone injectable formulations in lyophilized powder form. Chinese patent CN103705472A discloses sodium tanshinone powder for injection and its preparation method, mentioning the addition of antioxidants such as vitamin C for protection during the formulation process. However, simply adding antioxidants cannot guarantee long-term stability; this patent prepares it as a lyophilized powder. Meanwhile, existing studies have attempted to prepare lyophilized sodium tanshinone for injection (Zhang Jinjin et al., Safety Pharmacological Study of Sodium Tanshinone for Injection). Lyophilized powders require reconstitution before use, but the reconstitution process is cumbersome and time-consuming. Furthermore, the reconstituted solution often contains a large number of insoluble particles, increasing the safety risks of medication and limiting its widespread clinical application. Moreover, the reported sodium tanshinone injection formulations do not adequately improve product stability during actual production, and the cumbersome operation causes inconvenience in production.

[0006] Therefore, there is an urgent need and significant practical importance for a sodium tanshinone injection that is easy to operate, stable to heat, oxygen, and metal ions, and suitable for large-scale industrial production. Summary of the Invention

[0007] In view of the above problems, this invention aims to provide a sodium tanshinone injection and its industrial production method. By optimizing the preparation process, the precipitation problem of the injection is solved, providing a safer, more effective, stable, and long-term storage-compatible sodium tanshinone injection to meet the treatment needs of a wide range of patients.

[0008] During years of research on sodium tanshinone, the applicant discovered that sodium tanshinone injection is unstable and prone to precipitation during long-term storage, mainly due to metal ions, especially Fe. 3+ and Fe 2+ These metal ions may originate from the raw material and excipients of sodium tanshinone, or be introduced by equipment during the preparation process.

[0009] The above objectives are achieved through the following technical solutions:

[0010] A sodium tanshinone injection solution comprises sodium tanshinone, a stabilizer, and water for injection. The sodium tanshinone injection solution improves stability by reducing the metal ion content in the solution. The sodium tanshinone injection solution prepared according to this application contains Fe... 3+ The content is not higher than 0.5 μg / L, Fe 2+ The content is not higher than 1 μg / L.

[0011] Preparing the injection solution using sodium tanshinone, stabilizers, pH adjusters, and water for injection helps reduce the types of excipients in the injection solution, thereby improving its stability. The stabilizer in the injection solution has antioxidant and metal ion complexing effects, preventing sodium tanshinone from being oxidized during production and storage. It also forms complexes with metal ions, reducing the metal ion content in the sodium tanshinone injection solution, thus minimizing discoloration and improving its stability.

[0012] The pH adjuster is at least one of inorganic acid, organic acid, and inorganic base.

[0013] For example, the inorganic acid is one of hydrochloric acid, phosphoric acid, sulfuric acid, and acid buffer salt; preferably one of hydrochloric acid, phosphoric acid, and sulfuric acid, and more preferably hydrochloric acid.

[0014] For example, the inorganic base is one of sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, and alkaline buffer salt; preferably, it is one of sodium hydroxide, potassium hydroxide, sodium carbonate, and sodium bicarbonate.

[0015] The sodium tanshinone injection of the present invention comprises sodium tanshinone and a stabilizer, with concentrations of 1 mg / mL to 500 mg / mL and 0.02 mg / mL to 250 mg / mL, respectively. The stabilizer concentration is 1 / 2 to 1 / 50 (preferably 1 / 5 to 1 / 40, more preferably 1 / 10 to 1 / 30) of the sodium tanshinone concentration. The pH value of the injection is 1.0 to 6.0 (preferably 1.5 to 5.0, more preferably 2.0 to 4.0). The sodium tanshinone injection contains Fe... 3+ The content is not higher than 0.5 μg / L (preferably not higher than 0.25 μg / L, more preferably not higher than 0.1 μg / L); Fe 2+ The content is not higher than 1 μg / L (preferably not higher than 0.5 μg / L, more preferably not higher than 0.2 μg / L).

[0016] The stabilizer is disodium edetate or calcium sodium edetate. Adding disodium edetate or calcium sodium edetate as an antioxidant and metal ion chelating agent can play an antioxidant role and chelate metal ions in the injection solution.

[0017] When the pH value of the injection solution is below 1.0 or above 6.0, it will affect the stability of sodium tanshinone. Therefore, the present invention requires the pH value of the injection solution to be between 1.0 and 6.0.

[0018] Preferably, the tanshinone sodium injection of the present invention contains tanshinone sodium and a stabilizer, with concentrations of 2 mg / mL to 250 mg / mL and 0.05 mg / mL to 50 mg / mL, respectively.

[0019] More preferably, the sodium tanshinone injection of the present invention comprises sodium tanshinone and a stabilizer, with concentrations of 5 mg / mL to 50 mg / mL and 0.3 mg / mL to 5 mg / mL, respectively.

[0020] To ensure the stability and safety of sodium tanshinone injection during long-term storage, this invention addresses the Fe content in sodium tanshinone injection. 3+ and Fe 2+ Strict requirements were set for the content of Fe, which was used as a key quality control indicator. Therefore, this invention explores an industrial production method that avoids the introduction of Fe during the preparation process. 3+ and Fe 2+ This method minimizes metal ion contamination; nitrogen purging protects the solution from oxidation; and the addition of disodium edetate or calcium sodium edetate effectively prevents oxidation and chelates metal ions during long-term storage, further enhancing the stability and safety of tanshinone sodium injection. Through the comprehensive application of these measures, this invention fundamentally solves the precipitation problem of tanshinone sodium injection during long-term storage, ensuring its quality and safety. This innovative industrial production method not only enhances the product's market competitiveness but also provides patients with a more reliable and effective treatment option.

[0021] The following is the technical solution of the production method of the present invention:

[0022] This invention provides an industrial production method for the aforementioned sodium tanshinone injection, comprising the following steps:

[0023] (1) Take an acid-resistant mixing tank, inject water for injection into the mixing tank, add the prescribed amount of sodium tanshinone and stabilizer, and stir until all solid components are dissolved; wherein the amount of stabilizer added is 1 / 2 to 1 / 50 of the amount of sodium tanshinone added;

[0024] (2) Take a sample from the preparation tank and test the pH value of the solution. Adjust the pH value of the solution to 1.5-5.0 (preferably 2.0-4.0) using a pH adjuster. Add 0.1% (w / v) of medicinal charcoal and stir and adsorb at 60°C for 10-20 minutes. Then, filter the solution through a 1.0μm titanium filter to remove the charcoal. After removal of the charcoal, take a sample and test the concentration of sodium tanshinone in the solution.

[0025] (3) The decarbonized and filtered drug solution is transferred to a dilution tank; in the dilution tank, according to the central control test results, water for injection is added, and the pH value is adjusted if necessary, so that the concentration of sodium tanshinone is 1mg / mL to 500mg / mL (preferably 2mg / mL to 250mg / mL, more preferably 5mg / mL to 50mg / mL), the concentration of stabilizer is 0.02mg / mL to 250mg / mL (preferably 0.05mg / mL to 50mg / mL, more preferably 0.3mg / mL to 5mg / mL), and the pH value of the drug solution is 1.0 to 6.0 (preferably 1.5 to 5.0, more preferably 2.0 to 4.0);

[0026] (4) The drug solution in the dilution tank is filtered through a 0.45μm filter and two 0.22μm filters in sequence and then transferred to a buffer tank and filled under nitrogen flow.

[0027] (5) The drug solution in step (4) is sterilized at 121℃ for 15 min, inspected by light, and packaged to obtain the sodium tanshinone injection of the present invention.

[0028] In steps (1) to (4), the entire process from solution preparation to filling is protected with nitrogen.

[0029] Preferably, the acid-resistant mixing tank in step (1) is a mixing tank treated with fluoroplastic coating.

[0030] The stabilizer in step (1) is disodium edetate or sodium calcium edetate.

[0031] In step (2), the pH adjuster is preferably a 10 mol / L hydrochloric acid solution.

[0032] Nitrogen purging is used to prevent sodium tanshinone from coming into contact with oxygen, thus effectively improving the stability of the drug.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] 1. The production method of sodium tanshinone injection of the present invention is highly industrially operable, and the resulting injection has good stability. It does not require special storage conditions and can effectively avoid quality changes such as clarity, visible foreign matter, purity of active ingredients and related indicators caused by long-term storage of the injection, thus providing effective protection for the safety and clinical efficacy of the product.

[0035] 2. The tanshinone sodium injection of this invention is a preparation that can be used directly in clinical practice. It can be administered intravenously or prepared as an infusion solution, making it convenient to use. The preparation prepared by this process fully meets the stability and safety requirements for the preparation of injections and can be used in large quantities clinically.

[0036] 3. The sodium tanshinone injection of this invention can be used clinically by direct intravenous injection or prepared as an infusion for intravenous drip, which is convenient and quick. The addition of pH adjusting substances and stabilizers ensures the stability of the injection, allowing for long-term storage, safe use, and guaranteed quality. The added pH adjusting substances and stabilizers are at the concentrations permitted for intravenous drip administration, ensuring safety and reliability. It can be used to treat cardiovascular and cerebrovascular diseases, and has the effects of preventing and treating coronary microvascular diseases, anti-anginal heart disease angina pectoris, anti-atherosclerosis, protecting myocardial cells, and preventing ischemic damage to the heart and brain.

[0037] 4. The sodium tanshinone injection of the present invention is an injection that can be directly injected intravenously or prepared as an infusion for administration into the human body. It is isotonic with human blood and body fluids and will not cause hemolysis or other side effects, making it safe and reliable.

[0038] 5. The stabilizer added to the sodium tanshinone injection of this invention, disodium edetate or sodium calcium edetate, is a clinically permitted excipient for injection. It is safe and has no side effects. It can play an antioxidant role and chelate metal ions in the injection, ensuring the stability of the preparation and preventing changes in drug properties that may cause toxic side effects. It can be used more safely in clinical settings.

[0039] 6. In the industrial production and filling process, the sodium tanshinone injection of the present invention is filled with nitrogen gas, which avoids contact with oxygen, effectively prevents oxidation reaction, and also avoids the toxic side effects caused by drug oxidation, thus ensuring its safety.

[0040] 7. In the industrial production of the sodium tanshinone injection of this invention, the mixing tank is treated with a fluoroplastic coating, which effectively prevents the dissolution of iron ions from stainless steel under acidic conditions, thus controlling the amount of Fe metal ions in the injection. 3+ The content is not higher than 0.5 μg / L (preferably not higher than 0.25 μg / L, more preferably not higher than 0.1 μg / L), Fe 2+ The content is not higher than 1 μg / L (preferably not higher than 0.5 μg / L, more preferably not higher than 0.2 μg / L), which ensures its stability.

[0041] 8. Compared with the existing tanshinone-containing traditional Chinese medicine injection on the market, the present invention has good stability.

[0042] 9. Safety Tests of the Tanshinone Sodium Injection of this Invention: According to the national new drug registration classification, this invention is a new drug, and there are no marketed products of this invention worldwide. Therefore, this preparation is currently a blank in the global market, with broad market potential. We conducted safety tests according to relevant requirements: the results of the rabbit muscle irritation test showed that the product is non-irritating; the hemolysis and agglutination test showed that the product has no hemolytic or coagulating properties. These tests prove that the product is safe and reliable for clinical use. The product was subjected to accelerated testing for 6 months and long-term testing for 12 months, and the content was determined. No significant changes were observed in the stability results, indicating that the product has good stability under these process conditions. Attached Figure Description

[0043] Figure 1. Quadriceps femoris muscle of rabbits in the experimental group: negative control side (left) and test substance side (right).

[0044] Figure 2. Quadriceps femoris muscle of rabbits in the control group: negative control side (left) and test substance side (right).

[0045] Figure 3. Pathological section of the negative control lateral muscle fiber of the experimental group rabbits (HE staining, x100).

[0046] Figure 4. Pathological section of the lateral muscle fiber of the test sample from the experimental group rabbits (HE staining, x100).

[0047] Figure 5. Pathological section of the negative control side muscle fiber of the control group rabbit (HE staining, x100).

[0048] Figure 6. Pathological section of the test material lateral muscle fiber of the control group rabbit (HE staining, x100).

[0049] Figure 7. Hemolytic test of sodium tanshinone injection. Detailed Implementation

[0050] The present invention will now be described in detail with reference to specific embodiments. These embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several adjustments and improvements without departing from the concept of the present invention. These all fall within the scope of protection of the present invention.

[0051] QS is short for appropriate amount. The acid-resistant mixing tank in the example is a mixing tank treated with fluoroplastic coating.

[0052] Example 1: Preparation of Tanshinone Sodium Injection (1 mL: 5 mg)

[0053] prescription:

[0054] Preparation method:

[0055] (1) Take an acid-resistant mixing tank, inject 3000mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of sodium edetate and sodium tanshinone into the mixing tank in sequence, and purge the drug solution with nitrogen for protection. Stir for 30 minutes until completely dissolved.

[0056] (2) Sample and test the pH value. Adjust the pH value to 2.5 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 10 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of tanshinone sodium and the pH value of the solution were determined in the central control: the concentration of tanshinone sodium was 16.67 mg / ml, and the pH value was 2.5.

[0057] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0058] In the dilution container, add water for injection to the required amount according to the central control test results.

[0059] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0060] (5) The final filling solution is sterilized at 121℃ for 15 minutes, inspected by light, and packaged to obtain tanshinone sodium injection (1mL: 5mg).

[0061] Example 2: Preparation of Tanshinone Sodium Injection (5 mL: 100 mg)

[0062] prescription:

[0063] Preparation method:

[0064] (1) Take an acid-resistant mixing tank, inject 3000mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of sodium edetate and sodium tanshinone into the mixing tank in sequence, and purge the drug solution with nitrogen for protection. Stir for 30 minutes until completely dissolved.

[0065] (2) Sample and test the pH value. Adjust the pH value to 3.0 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 10 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of tanshinone sodium and the pH value of the solution were determined in the central control: the concentration of tanshinone sodium was 66.67 mg / ml, and the pH value was 3.0.

[0066] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0067] In the dilution container, add water for injection to the required amount according to the central control test results.

[0068] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0069] (5) The final filling solution is sterilized at 121℃ for 15 min, inspected by light, and packaged to obtain tanshinone sodium injection (5mL: 100mg).

[0070] Example 3: Preparation of Tanshinone Sodium Injection (5 mL: 200 mg)

[0071] prescription:

[0072] Preparation method:

[0073] (1) Take an acid-resistant mixing tank, inject 3000mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of disodium edetate and sodium tanshinone into the mixing tank in sequence, and protect the drug solution with nitrogen. Stir for 30 minutes until completely dissolved.

[0074] (2) Sample and test the pH value. Adjust the pH value to 3.0 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 10 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of sodium tanshinone and the pH value of the solution were determined in the central control: the concentration of sodium tanshinone was 133.33 mg / ml, and the pH value was 3.0.

[0075] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0076] In the dilution container, add water for injection to the required amount according to the central control test results.

[0077] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0078] (5) The final filling solution is sterilized at 121℃ for 15 minutes, inspected by light, and packaged to obtain tanshinone sodium injection (5mL: 200mg).

[0079] Example 4: Preparation of Tanshinone Sodium Injection (10 mL: 100 mg)

[0080] prescription:

[0081] Preparation method:

[0082] (1) Take an acid-resistant mixing tank, inject 12000mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of disodium edetate and sodium tanshinone into the mixing tank in sequence, and purge the drug solution with nitrogen for protection. Stir for 30 minutes until completely dissolved.

[0083] (2) Sample and test the pH value. Adjust the pH value to 2.5 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 15 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of tanshinone sodium and the pH value of the solution were determined in the central control: the concentration of tanshinone sodium was 33.33 mg / ml, and the pH value was 2.5.

[0084] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0085] In the dilution container, add water for injection to the required amount according to the central control test results.

[0086] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0087] (5) The final filling solution is sterilized at 121℃ for 15 min, inspected by light, and packaged to obtain tanshinone sodium injection (10mL: 100mg).

[0088] Example 5: Preparation of Tanshinone Sodium Injection (4 mL: 100 mg)

[0089] prescription:

[0090] Preparation method:

[0091] (1) Take an acid-resistant mixing tank, inject 2400mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of disodium edetate and sodium tanshinone into the mixing tank in sequence, and purge the drug solution with nitrogen for protection. Stir for 30 minutes until completely dissolved.

[0092] (2) Sample and test the pH value. Adjust the pH value to 3.0 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 20 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of tanshinone sodium and the pH value of the solution were determined in the central control: the concentration of tanshinone sodium was 83.33 mg / ml, and the pH value was 3.0.

[0093] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0094] In the dilution container, add water for injection to the required amount according to the central control test results.

[0095] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0096] (5) The final filling solution is sterilized at 121℃ for 15 min, inspected by light, and packaged to obtain tanshinone sodium injection (4mL: 100mg).

[0097] Example 6: Preparation of Tanshinone Sodium Injection (1 mL: 5 mg)

[0098] prescription:

[0099] Preparation method:

[0100] (1) Take an acid-resistant mixing tank, inject 3000mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of disodium edetate and sodium tanshinone into the mixing tank in sequence, and protect the drug solution with nitrogen. Stir for 30 minutes until completely dissolved.

[0101] (2) Sample and test the pH value. Adjust the pH value to 3.0 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 10 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of tanshinone sodium and the pH value of the solution were determined in the central control: the concentration of tanshinone sodium was 16.67 mg / ml, and the pH value was 3.0.

[0102] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0103] In the dilution container, add water for injection to the required amount according to the central control test results.

[0104] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0105] (5) The final filling solution is sterilized at 121℃ for 15 minutes, inspected by light, and packaged to obtain tanshinone sodium injection (1mL: 5mg).

[0106] Example 7: Preparation of Tanshinone Sodium Injection (5 mL: 250 mg)

[0107] prescription:

[0108] Preparation method:

[0109] (1) Take an acid-resistant mixing tank, inject 3000mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of sodium edetate and sodium tanshinone into the mixing tank in sequence, and purge the drug solution with nitrogen for protection. Stir for 30 minutes until completely dissolved.

[0110] (2) Sample and test the pH value. Adjust the pH value to 3.0 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 20 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of tanshinone sodium and the pH value of the solution were determined in the central control: the concentration of tanshinone sodium was 166.66 mg / ml, and the pH value was 3.0.

[0111] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0112] In the dilution container, add water for injection to the required amount according to the central control test results.

[0113] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0114] (5) The final filling solution is sterilized at 121℃ for 15 minutes, inspected by light, and packaged to obtain tanshinone sodium injection (5mL: 250mg).

[0115] Example 8: Preparation of Tanshinone Sodium Injection (5 mL: 5 mg)

[0116] prescription:

[0117] Preparation method:

[0118] (1) Take an acid-resistant mixing tank, inject 3000mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of sodium edetate and sodium tanshinone into the mixing tank in sequence, and purge the drug solution with nitrogen for protection. Stir for 30 minutes until completely dissolved.

[0119] (2) Sample and test the pH value. Adjust the pH value to 3.0 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 15 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of tanshinone sodium and the pH value of the solution were determined in the central control: the concentration of tanshinone sodium was 3.33 mg / ml, and the pH value was 3.0.

[0120] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0121] In the dilution container, add water for injection to the required amount according to the central control test results.

[0122] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0123] (5) The final filling solution is sterilized at 121℃ for 15 minutes, inspected by light, and packaged to obtain tanshinone sodium injection (5mL: 5mg).

[0124] Example 9: Preparation of Tanshinone Sodium Injection (5 mL: 2500 mg)

[0125] prescription:

[0126] Preparation method:

[0127] (1) Take an acid-resistant mixing tank, inject 3000mL of water for injection into the mixing tank, and at the same time open the nitrogen valve and turn on the stirring switch. Add the prescribed amount of disodium edetate and sodium tanshinone into the mixing tank in sequence, and protect the drug solution with nitrogen. Stir for 30 minutes until completely dissolved.

[0128] (2) Sample and test the pH value. Adjust the pH value to 3.0 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 20 min, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of tanshinone sodium and the pH value of the solution were determined in the central control: the concentration of tanshinone sodium was 16.67 mg / ml, and the pH value was 3.0.

[0129] (3) Open the receiving valve of the diluent tank to deliver the medicine into the diluent tank, and at the same time open the nitrogen valve to fill with nitrogen until the filling is completed;

[0130] In the dilution container, add water for injection to the required amount according to the central control test results.

[0131] (4) Open the inlet valve of the buffer tank. The medicine solution in the dilution tank is transported to the buffer tank after being filtered by a 0.45μm filter and two 0.22μm filter cartridges. The buffer tank is then filled and sealed under nitrogen flow.

[0132] (5) The final filling solution is sterilized at 121℃ for 15 minutes, inspected by light, and packaged to obtain tanshinone sodium injection (5mL: 500mg).

[0133] Comparative Example 1: The existing Chinese medicine Danshen injection containing tanshinone on the market was manufactured by Lei Yunshang Pharmaceutical Co., Ltd.

[0134] Comparative Example 2: Prepared according to the following method:

[0135] Take a stainless steel mixing tank and pour 3000 mL of water for injection into it. Simultaneously, open the nitrogen valve and turn on the stirrer. Add 200 g of sodium tanshinone to the mixing tank, purging the solution with nitrogen and stirring for 30 minutes until completely dissolved. Sample and test the pH value, adjusting it to 3.0 with 10 mol / L hydrochloric acid solution. Add 0.1% (w / v) pharmaceutical charcoal. After adsorption by stirring at below 60℃ for 10 minutes, filter through a 1.0 μm titanium filter to remove the charcoal. The concentration of sodium tanshinone and the pH value of the solution were measured in the central control unit: sodium tanshinone concentration was 66.67 mg / mL, and pH value was 3.0. Open the receiving valve of the dilution tank to transfer the solution into the dilution tank, simultaneously purging with nitrogen until filling is complete. Add water for injection to the dilution tank according to the central control unit's test results to prepare the full volume (10000 mL). Open the inlet valve of the buffer tank. The drug solution in the dilution tank is transferred to the buffer tank after being filtered through a 0.45μm filter and two 0.22μm filters. The solution is then filled under nitrogen flow. Finally, the filled solution is sterilized at 121℃ for 15 minutes, inspected by light, and packaged to obtain tanshinone sodium injection (5mL: 100mg).

[0136] The beneficial effects of the present invention are demonstrated through specific experiments below. The injection solutions obtained in the examples and comparative examples were used as samples for the experiments.

[0137] Experiment 1: Finished product inspection results of the sodium tanshinone injection of this invention

[0138] The quality of the sample samples in the examples was tested according to the 2020 edition of the Chinese Pharmacopoeia. Inductively coupled plasma mass spectrometry (ICP-MS) was used to determine the Fe ion content in the sample samples. 3+ and Fe 2+ The content of tanshinone sodium and stabilizer was determined by high performance liquid chromatography (HPLC), and the results are shown in Table 1. It is evident that the tanshinone sodium injection produced by this invention is of high quality.

[0139] Table 1. Test results of the finished product of the sodium tanshinone injection of this invention.

[0140] Experiment 2: Stability test of the sodium tanshinone injection of the present invention

[0141] Experimental procedure: The samples of Examples 1-5 and Comparative Examples 1-2 were subjected to accelerated testing (40℃±2℃, relative humidity 75%±5%). Samples were taken and tested after 3 months and 6 months respectively, and compared with the samples at 0 days.

[0142] The experimental results are shown in Table 2.

[0143] Table 2 Accelerated test stability results

[0144] In Table 2, the active ingredient in Examples and Comparative Example 2 refers to sodium tanshinone, and the active ingredient in Comparative Example 1 refers to tanshinone.

[0145] As can be seen from the results in Table 2, the sodium tanshinone injection prepared by the formulation and process of this invention (samples of Examples 1-5) maintains extremely high stability. After storage for a certain period of time, the purity and clarity of sodium tanshinone in the sodium tanshinone injection of this invention are very stable; compared with Examples 1-5, the Fe in Comparative Examples 1-2 3+ Content (ng / L) and Fe 2+ The content (ng / L) was significantly higher than that of the experimental group, and the color deepened to varying degrees after 3 and 6 months. The purity of tanshinone or sodium tanshinone continuously decreased, while the impurity content continuously increased. This indicates that the stability of the sodium tanshinone injection of this invention is better than that of sodium tanshinone injection prepared with similar formulations and preparation processes (Comparative Example 2), and better than that of tanshinone injection prepared with similar products on the market (Comparative Example 1). The sodium tanshinone injection of this invention has significant advantages.

[0146] Experiment 3: The sodium tanshinone injection of the present invention at different concentrations of Fe 3+ and Fe 2+ Medium stability test

[0147] To further verify Fe 3+ Content and Fe 2+ The effect of content on the stability of sodium tanshinone injection was investigated. 500 ml of sodium tanshinone injection from Example 2 was divided into five experimental groups, each containing 100 ml. Different amounts of FeCl2 and FeCl3 were added to each group, resulting in the AE group injection. The AE group injection samples were subjected to accelerated testing (40℃±2℃, relative humidity 75%±5%), and samples were taken and tested after 3 months and 6 months, respectively, and compared with the 0-day sample. The experimental results are shown in Table 3.

[0148] Table 3. Accelerated test stability results

[0149] In Table 3, the active ingredient of the AE group injection is sodium tanshinone.

[0150] As can be seen from the results in Table 3, the sodium tanshinone injection (sample of Example 2) prepared by the formulation and process of this invention has good Fe content. 3+ Content (0.5 μg / L) and Fe 2+Within a certain range (1 μg / L), the sodium tanshinone content (groups AD and D) maintained extremely high stability. After storage for a certain period of time, the purity and clarity of the sodium tanshinone injection were very stable. In group E, Fe... 3+ Content (ng / L) higher than 0.5 μg / L and Fe 2+ When the content (ng / L) is higher than 1 μg / L, add Fe 3+ and Fe 2+ Subsequently, after 3 and 6 months, the color continued to deepen, visible foreign matter continued to increase, the turbidity of the solution continued to increase, and the purity of sodium tanshinone continued to decrease.

[0151] Experiment 4: Muscle irritation test of the sodium tanshinone injection of the present invention.

[0152] Two qualified rabbits were selected and divided into an experimental group and a control group. The animals were used in a self-controlled manner, with the right quadriceps femoris muscle serving as the test substance administration side. The experimental group received the sample from Example 2, and the control group received the sample from Comparative Example 2. The left quadriceps femoris muscle served as a negative control side, receiving 5% glucose injection. The drug was administered intramuscularly once daily for a total of three times, with a dosage of 1 mL per rabbit. The injection site of the quadriceps femoris muscle was visually observed before each administration and at 24 and 48 hours after administration. At the end of the observation period, the animals were sacrificed, and the quadriceps femoris muscle was dissected, longitudinally incised, and the muscle stimulation response at the injection site was observed. The dissected muscle tissue was prepared for histopathological examination. The irritant effect of the injection on the animal muscle was determined by a combination of visual observation and histopathological examination. The results are as follows:

[0153] The above experimental results showed that no obvious abnormalities were observed in the quadriceps femoris muscles of all animals before administration. No other obvious muscle stimulation symptoms were observed during the administration period and at the end of the observation period. After the observation period, no obvious abnormalities were observed in the dissected quadriceps femoris muscles of the rabbits in both the experimental and control groups, as well as in the test sample side (see Figures 1-2). Histopathological examination showed no obvious abnormalities in the muscle fibers of the negative control side and the test sample side of the experimental and control rabbits (see Figures 3-5); the test sample side of the control rabbits showed degeneration and necrosis of muscle fibers, inflammatory cell infiltration, interstitial congestion and edema, and inflammatory cell infiltration (see Figure 6). The experimental results indicate that the safety of the tanshinone sodium injection of the present invention is better than that of the tanshinone sodium injection prepared with similar formulations and preparation processes (Comparative Example 2), and the tanshinone sodium injection of the present invention has significant advantages.

[0154] Experiment 5: Hemolytic activity test of the sodium tanshinone injection of this invention.

[0155] Referring to the General Chapters of Part IV of the 2020 edition of the Chinese Pharmacopoeia, this study observed whether tanshinone sodium injection caused hemolysis and erythrocyte aggregation in vitro. Rabbit erythrocytes were selected as the experimental system for an in vitro hemolysis test using a test tube method. The injection solution from Example 4 was prepared into a 1 mg / ml tanshinone sodium injection solution as the test solution. Eight clean, stoppered test tubes were used and numbered: tubes 1-3 were the test tubes (i.e., the tanshinone sodium injection group, repeated three times); tube 4 was the test control tube; tubes 5 and 6 were negative control tubes; and tubes 7 and 8 were positive control tubes. After incubation for 24 hours, no hemolysis or aggregation was observed in the tanshinone sodium injection group. The supernatant in the test tubes remained clear, and the number of cells sedimented at the bottom of the tubes was not significantly different from that in the negative control. Upon shaking the test tubes at the end of the test, the precipitated blood cells were dispersed. The positive control tubes remained in a hemolytic state throughout. The experimental system was valid, and the results were reliable (see Figure 7).

[0156] Experimental results show that the sodium tanshinone injection of the present invention has high safety and significant advantages.

Claims

1. A sodium Danshensu injection, characterized in that, The solution contains sodium tanshinone and a stabilizer at concentrations of 1 mg / mL to 500 mg / mL and 0.02 mg / mL to 250 mg / mL, respectively, and the stabilizer concentration is 1 / 2 to 1 / 50 of the sodium tanshinone concentration. The pH of the injection solution is 1.0 to 6.

0. The stabilizer is sodium edetate or sodium calcium edetate.

2. The injection fluid of claim 1, wherein, Fe in sodium tanshinone injection 3+ Content not exceeding 0.5 μg / L; Fe 2+ The content is not higher than 1 μg / L.

3. The injectable solution according to claim 1, characterized in that, Fe in sodium tanshinone injection 3+ Content not exceeding 0.25 μg / L; Fe 2+ The content is not higher than 0.5 μg / L.

4. The injectable solution according to claim 1, characterized in that, Fe in sodium tanshinone injection 3+ Content not exceeding 0.1 μg / L; Fe 2+ The content is not higher than 0.2 μg / L.

5. The injection solution according to any one of claims 1 to 4, characterized in that, The concentration of sodium tanshinone is 2 mg / mL to 250 mg / mL, and the concentration of the stabilizer is 0.05 mg / mL to 50 mg / mL.

6. The injection fluid of claim 5, wherein, The concentration of sodium tanshinone is 5 mg / mL to 50 mg / mL, and the concentration of the stabilizer is 0.3 mg / mL to 5 mg / mL.

7. An industrial production method of the sodium tanshinone injection solution according to any one of claims 1 to 6, comprising the following steps: (1) Take an acid-resistant mixing tank, inject water for injection into the mixing tank, add the prescribed amount of sodium tanshinone and stabilizer, and stir until all solid components are dissolved; wherein the amount of stabilizer added is 1 / 2 to 1 / 50 of the amount of sodium tanshinone added; (2) Take a sample from the preparation tank and test the pH value of the solution. Adjust the pH value of the solution to the range of 2.0 to 4.0 using a pH adjuster. Add 0.1% (w / v) of medicinal charcoal and stir and adsorb at 60°C for 10 to 20 minutes. Then, filter the solution through a 1.0 μm titanium filter to remove the charcoal. After decarbonization, take a sample and test the concentration of sodium tanshinone and the pH value of the solution. (3) Transfer the decarbonized and filtered drug solution to the dilution tank; in the dilution tank, according to the central control test results, add water for injection, and adjust the pH value if necessary, so that the sodium tanshinone concentration is 1mg / mL~500mg / mL, the stabilizer concentration is 0.02mg / mL~250mg / mL, and the pH value of the drug solution is 1.0~6.

0. (4) The drug solution in the dilution tank is filtered through a 0.45μm filter and two 0.22μm filters in sequence and then transferred to a buffer tank and filled under nitrogen flow. (5) The drug solution in step (4) is sterilized at 121℃ for 15 min, inspected by light, and packaged to obtain the sodium tanshinone injection of the present invention. In steps (1) to (4), the entire process from solution preparation to filling is protected with nitrogen. The stabilizer is disodium edetate or sodium calcium edetate.

8. The industrial production process according to claim 7, characterized in that, in, The acid-resistant mixing tank mentioned in step (1) is a mixing tank treated with fluoroplastic coating.

9. The industrial production method according to claim 7, characterized in that, The pH adjuster is at least one of inorganic acid, organic acid, and inorganic base.

10. The industrial production method according to claim 9, characterized in that, The pH adjuster is one of hydrochloric acid, phosphoric acid, and sulfuric acid.

11. The industrial production method according to claim 10, characterized in that, The pH adjuster is a 10 mol / L hydrochloric acid solution.

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