Sodium thiosulfate injection as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a sodium thiosulfate injection as well as a preparation method and application thereof. The injection consists of sodium thiosulfate, potassium chloride, disodium hydrogen phosphate, phosphoric acid, sodium hydroxide and water for injection. In order to avoid reproduction harm caused by boric acid as a pH buffer solvent in the prior art, glycine and phosphate are screened as the pH buffer solvent, it is found that glycine as the pH buffer solvent is poor in storage stability and high in related substance index, and compared with disodium hydrogen phosphate and sodium dihydrogen phosphate as buffer solvents, the glycine has the advantages that the storage stability is high; disodium hydrogen phosphate and phosphoric acid are used as buffer solvents, so that the stability is better, related indexes are low, and the dosage of disodium hydrogen phosphate is further optimized. The sodium thiosulfate injection screened layer by layer is stable in property, free of reproductive toxicity, small in pain feeling, small in irritation, free of hemolytic risk accidentally and especially suitable for children to use.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a sodium thiosulfate injection, a preparation method thereof and an application Background Art

[0002] Sodium Thiosulfate Injection can be clinically used in combination with sodium nitrite for the treatment of acute cyanide poisoning. At home and abroad, sodium thiosulfate injection is used off-label for calciphylaxis. In addition, in China, sodium thiosulfate injection is also used off-label for urticaria and pruritus of hemodialysis patients. This injection can be used for adults, slowly injected intravenously at 12.5 - 25 g. If necessary, half or full dose can be repeated after 1 hour. It can also be used for children, with a pediatric intravenous injection of 250 - 500 mg / kg, once a day

[0003] In 2011, the sodium thiosulfate injection of Hope Pharmaceuticals was approved for marketing in the United States. Its English name is Sodium Thiosulfate Injection, with a specification of 50 ml: 12.5 g, included in the US Orange Book, and having the status of RLD (Reference Listed Drug) and RS (Reference Standard) preparations. Subsequently, it was successively marketed in many countries. The formulation of this preparation contains the excipient boric acid. Boric acid is a common pharmaceutical excipient, used as an antibacterial agent and a pH buffer, and is widely included in national pharmacopoeias and pharmaceutical excipient manuals. However, in recent years, some studies have suggested that there may be a risk of reproductive toxicity when it is used in children

[0004] On the other hand, the results of special safety experiments on animals of the original research injection of sodium thiosulfate show that hemolysis is likely to occur after injection, and there are problems such as greater irritation, indicating that injection pain, irritation and other discomforts are likely to occur during clinical use, which will reduce the acceptability of patients including children to inject this product

[0005] In addition, the aqueous solution of sodium thiosulfate is relatively sensitive to oxygen and high temperature, etc. After sterilization and long-term storage, especially during the accelerated retention sample process, it is unstable, and problems such as increased impurities and decreased content are likely to occur. These changes in the drug may also be related to the adverse reactions that occur during clinical use

[0006] In summary, developing a sodium thiosulfate injection with stable liquid properties, especially without the risk of reproductive toxicity when used in children, without the risk of hemolysis during injection, with little pain and little irritation during injection is an urgent problem to be solved in this field Summary of the Invention

[0007] Aiming at the defects of the existing technology, the present invention provides a safe and stable sodium thiosulfate injection and a preparation method thereof to solve the problems in the existing technology

[0008] The technical problem to be solved by the present invention is that in the prior art, sodium thiosulfate injection has reproductive toxicity, high irritation, insecurity, and instability. The present invention provides a sodium thiosulfate injection and a preparation method thereof, and surprisingly finds that the provided sodium thiosulfate injection does not produce a hemolysis risk.

[0009] In a first aspect, in order to achieve the above object, the present invention provides a sodium thiosulfate injection, comprising sodium thiosulfate, a chloride salt, a pH buffer phosphate, and a pH regulator.

[0010] In some embodiments, the chloride salt is selected from sodium chloride or potassium chloride, and preferably the chloride salt is potassium chloride.

[0011] In some embodiments, the pH regulator is selected from sodium hydroxide, potassium hydroxide, sodium acetate, sodium bicarbonate, sodium citrate, sodium tartrate, hydrochloric acid, boric acid, acetic acid, or phosphoric acid. Preferably, the pH regulator is sodium hydroxide and phosphoric acid.

[0012] In some embodiments, the pH buffer phosphate is a buffer composed of disodium hydrogen phosphate and phosphoric acid.

[0013] For the sodium thiosulfate injection provided by the present invention, the phosphate is 1-17.5 g / L of disodium hydrogen phosphate, preferably 3-12 g / L of disodium hydrogen phosphate, more preferably 7.5-10 g / L of disodium hydrogen phosphate, and an appropriate amount of phosphoric acid.

[0014] In some embodiments, the components of the sodium thiosulfate injection provided by the present invention are 100-500 g / L of sodium thiosulfate, 1-10 g / L of potassium chloride, 3-12 g / L of disodium hydrogen phosphate, an appropriate amount of phosphoric acid, and an appropriate amount of sodium hydroxide; preferably, 250 g / L of sodium thiosulfate, 4.4 g / L of potassium chloride, 7.5-10 g / L of disodium hydrogen phosphate, an appropriate amount of phosphoric acid, and an appropriate amount of sodium hydroxide.

[0015] In some embodiments, the components of the sodium thiosulfate injection provided by the present invention are 250 g / L of sodium thiosulfate, 4.4 g / L of potassium chloride, 8 g / L of disodium hydrogen phosphate, an appropriate amount of phosphoric acid, and an appropriate amount of sodium hydroxide.

[0016] In some embodiments, the pH value of the sodium thiosulfate injection is 7.5-9.5, and preferably the pH value is 8.0-8.5.

[0017] In some embodiments, the solvent of the sodium thiosulfate injection is distilled water, sterile water, or water for injection, and preferably water for injection.

[0018] In another preferred embodiment of the present invention, the small-volume sodium thiosulfate injection contains 1.0 g of sodium thiosulfate, 17.6 mg of chloride salt, 32.0 mg of pH buffer, an appropriate amount of pH regulator, and an appropriate amount of water for injection, with a pH value of 7.5 to 9.0, and the injection is prepared into 4 mL in total.

[0019] In another preferred embodiment of the present invention, the large-volume sodium thiosulfate injection contains 25 g of sodium thiosulfate, 440 mg of chloride salt, 800 mg of pH buffer, an appropriate amount of pH regulator, and an appropriate amount of water for injection, with a pH value of 7.5 to 9.0, and the injection is prepared into 100 mL in total.

[0020] In another preferred embodiment of the present invention, the large-volume sodium thiosulfate injection contains 12.5 g of sodium thiosulfate, 220 mg of chloride salt, 400 mg of pH buffer, an appropriate amount of pH regulator, and an appropriate amount of water for injection, with a pH value of 7.5 to 9.0, and the injection is prepared into 50 mL in total.

[0021] Second, the present invention provides a method for preparing the sodium thiosulfate injection of the first aspect, comprising the following steps:

[0022] (1) Add water for injection to the dispensing tank, introduce an inert gas into the dispensing tank, and control the dissolved oxygen.

[0023] (2) Add the prescribed amounts of potassium chloride and disodium hydrogen phosphate and stir to dissolve. After adjusting the pH value to 8.5 - 9.0 with the pH regulator phosphoric acid or sodium hydroxide, then add sodium thiosulfate to dissolve.

[0024] (3) Adjust the pH value to 7.5 - 9.0 with the pH regulator phosphoric acid or sodium hydroxide, and make up a small amount of water for injection to make up the volume.

[0025] (4) The liquid medicine is filtered through a filter element, filled, heat-sealed, and sterilized by high-pressure moist heat to obtain the sodium thiosulfate injection.

[0026] In some embodiments, the method for preparing the sodium thiosulfate injection of the present invention is as follows:

[0027] (1) Add water for injection to the dispensing tank, introduce an inert gas into the dispensing tank, so that the dissolved oxygen in the water for injection is within 0.5 ppm;

[0028] (2) After cooling to 20 - 40 °C, add the chloride salt and pH buffer to dissolve, and then adjust the pH value to 8.5 - 9.0 with the pH regulator phosphoric acid and add sodium thiosulfate;

[0029] (3) After adjusting the pH value to 7.5 - 9.0 with the pH regulator phosphoric acid or sodium hydroxide, make up the volume with water for injection.

[0030] (4) Filling and sterilization.

[0031] In a preferred embodiment of the present invention, after volume determination, the liquid medicine is filled and sealed by a blow-fill-seal integrated machine, filled with nitrogen, and the residual oxygen in the headspace of the plastic ampoule is controlled within 3%. The plastic ampoules are placed in a composite film bag or an aluminum-plastic bag with high barrier properties to oxygen and water vapor, etc.

[0032] In another preferred embodiment of the present invention, after volume determination, the liquid medicine is filled into a plastic infusion soft bag, filled with nitrogen, and the residual oxygen in the headspace of the soft bag is controlled within 5%. The plastic soft bag is placed in a composite film bag or an aluminum-plastic bag with high barrier properties to oxygen and water vapor, etc.

[0033] Furthermore, the conditions for high-temperature and humid heat sterilization are: 121 °C, 8 - 30 min, and more preferably 121 °C, F 0 ≥ 12.

[0034] In a third aspect, the present invention provides the sodium thiosulfate injection of the first aspect or the sodium thiosulfate injection prepared by the preparation method of the second aspect for treating or preventing diseases such as ototoxicity, cyanide poisoning, eye diseases, hearing loss, urticaria, skin pruritus, etc. in children or adults, or for calcification defense and disinfectants.

[0035] Preferably, the sodium thiosulfate injection of the first aspect of the present invention or the sodium thiosulfate injection prepared by the preparation method of the second aspect is used as a pediatric drug, such as a drug for diseases such as ototoxicity, cyanide poisoning, eye diseases, hearing loss, urticaria, skin pruritus, etc. in children, or for pediatric calcification defense and disinfectants. More preferably, it is used for pediatric ototoxicity.

[0036] Compared with the prior art, the present invention has the following technical effects:

[0037] (1) The pH regulator uses a non-boric acid pH buffer to avoid the risk of reproductive toxicity caused by the use of boric acid in the prior art, which is beneficial for pediatric drug use.

[0038] (2) Surprisingly, for the preferred phosphate pH buffer, the finished product prepared by the technical solution of the present invention not only has small related substances and stable properties, but also does not pose a risk of hemolysis during injection, has little injection irritation and little pain, which is beneficial for the use of pediatric patients clinically.

[0039] (3) The sodium thiosulfate injection of the present invention has a simple composition, fewer preparation process steps, and a feasible preparation method, which is suitable for large-scale production. Description of the Drawings

[0040] Figure 1 Shows the photos of each test tube in Comparative Example 2 after 3 hours of incubation;

[0041] Figure 2It shows the histopathology diagram of the right ear marginal vein of the negative control in Comparative Example 3 14 days after drug administration;

[0042] Figure 3 It shows the histopathology diagram of the left ear marginal vein of the test article group in Comparative Example 3 14 days after drug administration;

[0043] Figure 4 It shows the histopathology diagram of the left ear marginal vein of the reference preparation group in Comparative Example 3 14 days after drug administration. Detailed implementation manners

[0044] In order to more clearly and detailedly illustrate the object technical solutions of the present invention, the present invention will be further described through relevant embodiments below. The following embodiments are only for specifically illustrating the implementation methods of the present invention and do not limit the protection scope of the present invention.

[0045] Example 1

[0046] The prescription of the sodium thiosulfate injection of the present invention is as follows:

[0047] Component Dosage Sodium thiosulfate 12.5g Potassium chloride 220 mg Disodium hydrogen phosphate 400 mg Phosphoric acid Appropriate amount Sodium hydroxide Appropriate amount Water for injection 50 mL

[0048] Preparation process:

[0049] Add 90% of the total prescription water volume of injection water into the dispensing tank, introduce the inert gas nitrogen into the dispensing tank, and control the dissolved oxygen ≤ 0.5 ppm. Cool down to below 40°C, add the prescription amount of potassium chloride and disodium hydrogen phosphate and stir to dissolve. After adjusting the pH value to 8.5 - 9.0 with the pH value regulator phosphoric acid or sodium hydroxide, then add sodium thiosulfate to dissolve. Adjust the pH value to 7.5 - 9.0 with the pH value regulator phosphoric acid or sodium hydroxide, and make up a small amount of injection water to make up the volume. The liquid medicine is filtered through 0.45μm and 0.22μm filters, filled into a three-layer co-extruded infusion bag made of polypropylene material, the headspace is filled with nitrogen to control the residual oxygen within 0 - 5%, and heat-sealed. Add an outer bag with high barrier functions for oxygen and water vapor, and perform moist heat sterilization at 121°C, F0 ≥ 12.

[0050] Example 2

[0051] The prescription of the sodium thiosulfate injection of the present invention is as follows:

[0052] Component Dosage Sodium thiosulfate 25.0g Potassium chloride 440 mg Disodium hydrogen phosphate 800 mg Phosphoric acid Appropriate amount Sodium hydroxide Appropriate amount Water for injection 100 mL

[0053] Preparation process:

[0054] Add 90% of the total prescription water volume of injection water into the liquid dispensing tank, introduce inert gas nitrogen into the liquid dispensing tank, and control the dissolved oxygen ≤ 0.5 ppm. Cool down to below 40 °C, add the prescribed amount of potassium chloride and disodium hydrogen phosphate and stir to dissolve. After adjusting the pH value to 8.5 - 9.0 with pH regulator phosphoric acid or sodium hydroxide, then add sodium thiosulfate to dissolve. Adjust the pH value to 7.5 - 9.0 with pH regulator phosphoric acid or sodium hydroxide, and make up a small amount of injection water to make up the volume. The liquid medicine is filtered through 0.45 μm and 0.22 μm filters, filled into a three-layer co-extruded infusion bag made of polypropylene material, the headspace is filled with nitrogen to control the residual oxygen within 0 - 5%, and heat-sealed. Add an outer bag with high barrier functions for oxygen and water vapor, and sterilize by moist heat at 121 °C, F0 ≥ 12.

[0055] Example 3

[0056] The prescription of the sodium thiosulfate injection of the present invention is as follows:

[0057] Component Dosage Sodium thiosulfate 1.0g Potassium chloride 17.6 mg Disodium hydrogen phosphate 32.0 mg Phosphoric acid Appropriate amount Sodium hydroxide Appropriate amount Water for injection 4ml

[0058] Preparation process:

[0059] Add 90% of the total prescription water volume of injection water into the liquid dispensing tank, heat to boiling for 10 min, evacuate the headspace and fill with nitrogen 3 times and then maintain filling with nitrogen, control the dissolved oxygen ≤ 0.5 ppm. Cool down to 30 - 60 °C, add the prescribed amount of potassium chloride and boric acid and stir to dissolve. After adjusting the pH value to 7.8 - 8.7 with sodium hydroxide, then add sodium thiosulfate to dissolve. Adjust the pH value to 7.5 - 8.4 with sodium hydroxide, and make up a small amount of injection water to make up the volume. The liquid medicine is filtered through 0.45 μm and 0.22 μm filters, filled into a polymer plastic bottle, the headspace is filled with nitrogen to control the residual oxygen within 0 - 3%, and stopper and seal. Add a composite film bag or an aluminum-plastic bag with high barrier functions for oxygen and water vapor, etc. Sterilize by moist heat at 121 °C, F0 ≥ 12.

[0060] The present invention relates to the detection method of key quality indicators as follows, and the beneficial effects of the present invention are proved by the following comparative examples:

[0061] (1) Related substances:

[0062] Determine according to the high performance liquid chromatography method (General Principles 0512, Volume IV, Chinese Pharmacopoeia 2020 Edition).

[0063] For the test solution, accurately measure an appropriate amount of the preparation to be tested, and quantitatively dilute it with the mobile phase to prepare a solution containing about 2 mg of sodium thiosulfate per 1 mL.

[0064] For the control solution, accurately measure an appropriate amount of the test solution, and quantitatively dilute it with the mobile phase to prepare a solution containing about 20 μg of sodium thiosulfate per 1 mL.

[0065] System suitability solution: Weigh accurately appropriate amounts of sodium sulfite, sodium tetrathionate, and sodium thiosulfate reference substances, dissolve and dilute with the mobile phase to prepare a mixed solution containing about 2 mg of sodium thiosulfate, 3 μg of sodium sulfite, and 3 μg of sodium tetrathionate per 1 mL.

[0066] Chromatographic conditions: Use an octadecylsilyl silica gel bonded phase (C18, 4.6 mm × 150 mm, 5 μm or a chromatographic column with equivalent performance) as the filler; use a solution of disodium hydrogen phosphate (containing 0.0025 mol / L of tetrabutylammonium hydrogen sulfate, adjusted to pH 8.5 with 1 mol / L sodium hydroxide solution)-acetonitrile (75:25) as the mobile phase; the flow rate is 0.5 mL per minute; the column temperature is 20 °C; the detection wavelength is 212 nm; the injection volume is 20 μL; the injection tray temperature is 4 °C.

[0067] System suitability requirements: In the chromatogram of the system suitability solution, sulfite, thiosulfate, and tetrathionate should elute in sequence, and the resolution between the sulfite peak and the thiosulfate peak should meet the requirements.

[0068] Assay method: Accurately measure the test solution and the reference solution, and inject them into the liquid chromatograph respectively, and record the chromatogram.

[0069] Limit: After deducting the solvent peak in the chromatogram of the test solution, if there are impurity peaks, the peak areas of sodium sulfite and sodium tetrathionate shall not be greater than 0.15 times (0.15%) of the main peak area of the reference solution.

[0070] (2) Insoluble particles and visible foreign matters: For the detection of insoluble particles, refer to the method for the inspection of insoluble particles in the Chinese Pharmacopoeia (2020 Edition) (General Chapter 0903); for the detection of visible foreign matters, refer to the method for the inspection of visible foreign matters in the Chinese Pharmacopoeia (2020 Edition) (General Chapter 0904).

[0071] Information of the reference preparation (original developed preparation): A sodium thiosulfate injection, its content and composition information: In a 50 mL solution, there is 12.5 g of sodium thiosulfate, 2.8 mg of boric acid and 4.4 mg of potassium chloride per milliliter, and the pH value is adjusted with boric acid and sodium hydroxide, and the pH value is between 7.5 and 9.5.

[0072] Comparative Example 1 Screening of different buffers

[0073] (1) Screening experiment of glycine concentration

[0074] The prescriptions with different glycine concentrations selected are shown in the following table. Prepare samples of Prescription 1, 2, and 3 according to the following technological steps. Sterilize the samples of different prescriptions and conduct accelerated retention tests at 40 °C.

[0075] Component Prescription 1 Prescription 2 Prescription 3 Sodium thiosulfate 25.0g 25.0g 25.0g Potassium chloride 440 mg 440 mg 440 mg Glycine 400 mg 200 mg 100 mg Sodium hydroxide Appropriate amount Appropriate amount Appropriate amount Water for injection Add up to 100 mL Add up to 100 mL Add up to 100 mL

[0076] Preparation process:

[0077] Add 90% of the total amount of water for injection in the prescription to the liquid preparation tank, introduce inert gas nitrogen into the liquid preparation tank, and control the dissolved oxygen ≤ 0.5 ppm. Cool down to below 40 °C, add the potassium chloride and glycine in the prescription amount and stir to dissolve. After adjusting the pH value to 8.5 - 9.0 with pH regulator sodium hydroxide, then add sodium thiosulfate to dissolve. Adjust the pH value to 7.5 - 9.0 with pH regulator sodium hydroxide, and make up a small amount of water for injection to make up the volume. The liquid medicine is filtered through 0.45 μm and 0.22 μm filters, filled in a three-layer co-extruded infusion bag made of polypropylene material, and nitrogen is filled in the headspace to control the residual oxygen within 0 - 5%, and then heat-sealed. Add an outer bag with high-barrier oxygen and water vapor functions, and sterilize it by moist heat at 121 °C, F0 ≥ 12.

[0078] Keep samples selected from the above prescriptions with different glycine concentrations for retention inspection, and submit them for the following test indicators for inspection. The comparison results are as follows.

[0079] Table 1 Results of screening experiments with different glycine concentrations

[0080]

[0081] From the above results, it can be seen that for the original developed preparation during the 0-day inspection, the level of related substances is relatively high, there is a risk that the insoluble particles in the liquid medicine exceed the limit, and there are a small number of white dot-like visible foreign matters observed under the clarity meter. After 3 months at 40 °C in the accelerated test, the related substances in the reference preparation liquid medicine increase significantly, the number of insoluble particles increases sharply, and a large number of white dot-like visible foreign matters appear.

[0082] According to the above preparation process, samples are prepared using Prescription 2 and Prescription 3. When the glycine concentration in the prescription is relatively low, the pH value of the sample is unstable and shows a downward trend after sterilization. During the stability retention process of the sample, the pH value further decreases, the related substances show an increasing trend, and there are more white dot-like visible foreign matters. When the glycine concentration of Prescription 1 is relatively high, the pH value of the sample is stable and does not show a downward trend after sterilization. During the stability retention process of the sample, the pH value also does not show a downward trend. Moreover, during the accelerated retention process of Prescription 1, the level of insoluble particles is very low, and there is no risk of visible foreign matters. However, during the accelerated test process of Prescription 1, the related substances show an increasing trend.

[0083] (2) Screening experiment of sodium dihydrogen phosphate monohydrate + disodium hydrogen phosphate anhydrous combination

[0084] The screened prescriptions with different concentrations of sodium dihydrogen phosphate monohydrate + disodium hydrogen phosphate anhydrous combination are shown in the following table, and the process steps are described below. Sterilize the samples of different prescriptions and keep them for inspection at 40 °C in the accelerated test.

[0085] Component Prescription 1 Prescription 2 Prescription 3 Sodium thiosulfate 25.0g 25.0g 25.0g Potassium chloride 440 mg 440 mg 440 mg Sodium dihydrogen phosphate monohydrate 123 mg 246 mg 123 mg Disodium hydrogen phosphate anhydrous 16 mg 32 mg 16 mg Hydrochloric acid Appropriate amount Appropriate amount Appropriate amount Sodium hydroxide Appropriate amount Appropriate amount Appropriate amount pH value adjusted to 7.5-8.0 7.5-8.0 8.3 Water for injection Add up to 100 mL Add up to 100 mL Add up to 100 mL

[0086] Preparation process:

[0087] Dissolve anhydrous sodium thiosulfate into the buffer of sodium dihydrogen phosphate monohydrate and disodium hydrogen phosphate anhydrous, and adjust the pH to 7.5 - 8.0 or 8.3 with sodium hydroxide and hydrochloric acid. Filter twice through a 0.22 μm filter element. Fill the filtered solution into glass vials. Seal the vials with a film-coated rubber stopper and an aluminum cap, and crimp the cap. Autoclave the sealed and filled vials at 121 °C and 15 psi for at least 0.5 hours to sterilize the contents.

[0088] Keep samples of the preparations screened with the above different concentrations for retention inspection, and send them for inspection of the following detection indicators. The comparison results are as follows.

[0089] Table 2 Results of inspection of samples prepared with different prescriptions

[0090]

[0091]

[0092] Prescription 1 and Prescription 2 are combinations of sodium dihydrogen phosphate + disodium hydrogen phosphate, where the amount of sodium dihydrogen phosphate is relatively large and the amount of disodium hydrogen phosphate is relatively small. Sodium dihydrogen phosphate is acidic and the buffer system is acidic. Using this combination and following the above process, when the raw material sodium thiosulfate is put into the buffer system during the sample liquid preparation process, turbidity and precipitation occur. After the liquid medicine is filled into glass bottles, sealed and crimped, and autoclaved at 121 °C and 15 psi for 0.5 hours, the pH value of the sample drops significantly, and the related substances also increase a lot, far exceeding the quality standard limit.

[0093] Using Prescription 3 and following the above process to prepare the Prescription 3 preparation, after the liquid medicine is filled into glass bottles, sealed and crimped, and autoclaved at 121 °C and 15 psi for 0.5 hours, the pH value of the sample drops significantly, and the related substances also increase a lot; after accelerating and retaining samples at 40 °C for 3 months, needle-shaped crystals precipitate, affecting the safety of drug use. It shows that the quality of the samples prepared with the above prescription process and packaging materials is unstable.

[0094] (3) Screening experiment of disodium hydrogen phosphate anhydrous + phosphoric acid combination

[0095] The screened different concentration prescriptions of the disodium hydrogen phosphate anhydrous + phosphoric acid combination are shown in the following table, and the remaining process steps are the same as those in Example 1.

[0096] Sterilize the samples with different prescriptions and conduct retention inspection at 40 °C for acceleration.

[0097] Component Prescription 1 Prescription 2 Prescription 3 Prescription 4 Sodium thiosulfate 25.0g 25.0g 25.0g 25.0g Potassium chloride 440 mg 440 mg 440 mg 440 mg Disodium hydrogen phosphate 500 mg 700 mg 800 mg 900 mg Phosphoric acid Appropriate amount Appropriate amount Appropriate amount Appropriate amount Sodium hydroxide Appropriate amount Appropriate amount Appropriate amount Appropriate amount Water for injection Add up to 100 mL Add up to 100 mL Add up to 100 mL Add up to 100 mL

[0098] Send the samples screened with the above different phosphate concentrations for inspection of the following detection indicators. The comparison results are as follows.

[0099] Table 3 Results of different formulation screening experiments

[0100]

[0101]

[0102] Note: BRL represents below the limit of quantitation, and the value is very small.

[0103] According to the above process, for the preparation samples prepared with Formulation 1 and Formulation 2 at relatively low phosphate concentrations, the pH value of the samples was unstable and showed a downward trend after sterilization, and the pH value further decreased during the stability retention period of the samples. There was a tendency for the related substances to increase, and there were also a small number of white dot-like visible foreign matters.

[0104] When the phosphate concentrations of Formulation 3 and Formulation 4 were relatively high, the pH value of the samples was stable and did not show a downward trend after sterilization, and the pH value also did not show a downward trend during the stability retention period of the samples. Moreover, according to the above process, for the samples prepared with Formulation 3 - 4, during the accelerated retention period, the level of insoluble particles was very low, and there was no risk of visible foreign matters; compared with the reference preparation, the quality indicators such as related substances were better. According to the above test results, the preferred formulation concentration is 8.0 - 9.0 mg / mL.

[0105] Comparative Example 2: Comparison of in vitro hemolysis test

[0106] Through the in vitro hemolysis test of sodium thiosulfate injection solution, it was judged whether the test article sodium thiosulfate injection solution of Example 1 could cause reactions such as hemolysis and red blood cell aggregation, and compared with the control article (reference preparation), providing a reference for the clinical safe application of this preparation.

[0107] (1) Test method

[0108] Take 12 clean test tubes numbered 1 - 12. Add 2.5 mL of 2% red blood cell suspension to each tube.

[0109] Tube No. 1 is the negative control tube, add 2.5 mL of sodium chloride injection solution;

[0110] Tube No. 2 is the positive control tube, add 2.5 mL of sterile water for injection;

[0111] Tubes No. 3 - 7 are the test article tubes, add 2.4, 2.3, 2.2, 2.1 and 2.0 mL of sodium chloride injection solution respectively, and then add 0.1, 0.2, 0.3, 0.4 and 0.5 mL of the test article sodium thiosulfate injection solution stock solution (250 mg / mL) respectively;

[0112] Tubes No. 8 - 12 are reference preparation tubes. Add 2.4, 2.3, 2.2, 2.1, and 2.0 mL of sodium chloride injection respectively, and then add 0.1, 0.2, 0.3, 0.4, and 0.5 mL of the reference preparation sodium thiosulfate injection stock solution (250 mg / mL) respectively.

[0113] After mixing, immediately place it in a thermostatic water bath at 37 (±0.5) °C for light - protected incubation. Visually observe the solutions in each test tube at 15, 30, 45 min and 1, 2, 3 h after the start of incubation. Take pictures of the test tubes at the 3 - h incubation time point, see Figure 1 .

[0114] Centrifuge each tube at 1500 revolutions per minute for 10 minutes at 4 °C. Perform colorimetry at 545 nm on a UV - Vis spectrophotometer, zero with deionized water, measure the absorbance of the supernatant, read the absorbance value (OD) of each tube, and calculate the hemolysis percentage.

[0115] (2) Test results

[0116] Spectrophotometry:

[0117] The hemolysis percentages of the test article tubes with the test article solution volumes of 0.1 - 0.5 mL incubated for 3 h are 0.17%, 0.62%, 1.11%, 1.59%, and 3.59% in sequence;

[0118] The hemolysis percentages of the reference preparation tubes with the reference preparation solution volumes of 0.1 - 0.5 mL incubated for 3 h are 1.23%, 4.46%, 8.78%, 13.61%, and 16.88% in sequence.

[0119] The hemolysis percentages of the test tubes with the reference preparation volumes of 0.3 mL, 0.4 mL, and 0.5 mL are all greater than 5%, indicating hemolysis occurred. The hemolysis percentages of the remaining test tubes are all less than 5%, indicating no hemolysis occurred.

[0120] The absorbance and hemolysis test results of each group are shown in Table 4.

[0121] Table 4 Results of absorbance values and hemolysis rates

[0122] Test tube number Absorbance value ODt - ODnc ODpc - ODnc Hemolysis percentage (%) 1 0.0091 / / / 2 1.3394 / / / 3 0.0113 0.0022 1.3303 0.17 4 0.0173 0.0082 1.3303 0.62 5 0.0239 0.0148 1.3303 1.11 6 0.0303 0.0212 1.3303 1.59 7 0.0569 0.0478 1.3303 3.59 8 0.0255 0.0164 1.3303 1.23 9 0.0684 0.0593 1.3303 4.46 10 0.1259 0.1168 1.3303 8.78 11 0.1902 0.1811 1.3303 13.61 12 0.2336 0.2245 1.3303 16.88

[0123] Note: " / " represents not applicable.

[0124] (3) Test conclusion

[0125] Under the conditions of this test, the sodium thiosulfate injection samples of Examples 1 with different concentrations from No. 3 to No. 7 did not cause reactions such as hemolysis and red blood cell aggregation, and the results were negative. The reference preparations with lower concentrations from No. 8 to No. 9 did not cause hemolysis reactions, and the results were negative. The sodium thiosulfate injection of the reference preparations from No. 10 to No. 12 caused hemolysis reactions but did not cause red blood cell aggregation reactions, and the results were positive. The results after incubation of each test tube for 3 h are as Figure 1 shown.

[0126] Comparative Example 3: Comparison of rabbit vascular irritation test

[0127] Through the rabbit vascular irritation test of sodium thiosulfate injection, observe the local reactions of the rabbit's administration site (blood vessel) after contacting the sodium thiosulfate injection sample, and compare with the control (reference preparation) to provide a reference for the clinical safe application of this preparation.

[0128] Method

[0129] Twelve New Zealand white rabbits (half male and half female) were randomly stratified and grouped according to their body weights before grouping into 2 groups (sample group and reference preparation group), with 6 rabbits in each group, half male and half female.

[0130] The animals in the sample group were intravenously injected with the sodium thiosulfate injection stock solution (250 mg / mL) through the marginal ear vein of the left ear, and the animals in the reference preparation group were intravenously injected with the sodium thiosulfate injection stock solution (250 mg / mL) through the marginal ear vein of the left ear. The animals in both groups were intravenously injected with an equal volume of sodium chloride injection through the marginal ear vein of the right ear. The bilateral administration volume of all animals was 3 mL / kg. The administration rate was 120 mL / h, and the administration was a single dose.

[0131] During the test period, the animals were observed beside the cages every day. Before administration and 24 h, 48 h, and 72 h after administration, observe whether there are reactions such as congestion, swelling, and necrosis in the bilateral rabbit ears once, and then observe once every day. Three animals in each group were euthanized 72 hours after administration, and the remaining animals were euthanized 14 days after administration. Both ears were taken for gross necropsy and histopathological examination.

[0132] Results

[0133] Observation beside the cages:

[0134] During the test period, the general conditions, behaviors, and signs of all animals in the sample group and the reference preparation group were normal.

[0135] Observation of the administration site:

[0136] Negative control side: No irritation reactions were observed in all animals during the test period (the scores were all 0).

[0137] Test article side: At 24 h and 48 h after administration, among 6 animals, only 1 animal showed a stimulation reaction (score: 1), and no stimulation reaction was observed in the remaining animals (scores were all 0).

[0138] Reference preparation side: At 24 h after administration, among 6 animals, 4 animals successively showed stimulation reactions (scores: 1 - 2), and the stimulation reactions completely disappeared by the 5th day. No stimulation reaction was observed in the remaining animals (scores were all 0).

[0139] Note: Score the stimulation reaction according to the following table

[0140] Score Vascular wall irritation reaction 0 No obvious reaction 1 Mild congestion 2 Mild - moderate congestion, swelling 3 Moderate - severe congestion, swelling, ear drooping 4 Moderate - severe congestion, swelling, ear drooping, and mild - moderate necrosis 5 Moderate - severe congestion, swelling, ear drooping, and severe extensive necrosis

[0141] Histopathological examination:

[0142] At 72 h after administration, negative control group: Perivascular edema / hemorrhage was occasionally seen, considered to be caused by mechanical stimulation and vascular hemorrhage and extravasation due to injection puncture; Test article group: Drug irritation-related changes were seen in all animals, mainly manifested as endothelial cell hypertrophy and intimal degeneration of blood vessels, and thrombosis was also seen in some animals; Reference preparation group: Drug irritation-related changes were seen in all animals, mainly manifested as endothelial cell hypertrophy and intimal degeneration of blood vessels, and thrombosis, perivascular inflammation, degeneration and necrosis of blood vessel wall, and hemorrhage of blood vessel wall were also seen in some animals.

[0143] At 14 days after administration, the histopathological figures of the negative control group, test article group and reference preparation group are shown respectively as Figure 2 , Figure 3 and Figure 4 shown. The results shown in the figures are the microscopic pictures taken under a microscope after HE staining and magnification by 200 times. Negative control group: No obvious abnormality was seen in the ear marginal veins of all animals, Figure 2 which is a microscopic picture of a representative group in the negative control; Test article group: Drug irritation-related changes were seen in 2 animals, mainly manifested as endothelial cell hypertrophy and intimal degeneration of blood vessels, but both cases were less severe than those in the reference preparation group. No obvious abnormality was seen in the ear marginal veins of the other 4 animals, Figure 3 which is a microscopic picture of a representative group among the 4 animals without obvious abnormality in the test article group; Reference preparation group: Drug irritation-related changes were seen in all 6 animals, mainly manifested as thickening of blood vessel wall, and degeneration of blood vessel wall was also seen in some animals, Figure 4 which is a microscopic picture of a representative group among the 6 animals in the reference preparation group.

[0144] Under the conditions of this test, the vascular irritation of rabbits was investigated by single intravenous injection of the test article sodium thiosulfate injection and the reference preparation sodium thiosulfate injection through observation at the administration site and histopathological examination. Especially 14 days after drug withdrawal, the number of animals with irritation caused by the test article was significantly less than that of the reference preparation, and the degree of irritation was lighter.

[0145] Finally, it should be noted that the above are only embodiments of the present application, which are only used to illustrate the technical solutions of the present invention and are not used to limit the present application. For those skilled in the art, various changes and modifications can be made to the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included within the scope of the claims of the present application.

Claims

1. A sodium thiosulfate injection, characterized in that: Contains sodium thiosulfate, chloride salt, pH buffer phosphate, pH adjuster.

2. The sodium thiosulfate injection according to claim 1, characterized in that: The chloride salt is selected from sodium chloride or potassium chloride.

3. The sodium thiosulfate injection according to any one of claims 1-2, characterized in that: The pH adjuster is selected from sodium hydroxide, potassium hydroxide, sodium acetate, sodium bicarbonate, sodium citrate, sodium tartrate, hydrochloric acid, boric acid, acetic acid, and phosphoric acid. Preferably, the pH adjuster is selected from sodium hydroxide and phosphoric acid.

4. The sodium thiosulfate injection according to any one of claims 1 to 3, characterized in that: The pH buffer phosphate is a buffer composed of disodium hydrogen phosphate and phosphoric acid.

5. The sodium thiosulfate injection according to any one of claims 1 to 4, characterized in that: The phosphate is 1-17.5 g / L disodium hydrogen phosphate, preferably 3-12 g / L disodium hydrogen phosphate, more preferably 7.5-10 g / L disodium hydrogen phosphate, and an appropriate amount of phosphoric acid.

6. The sodium thiosulfate injection according to any one of claims 1 to 5, characterized in that: Sodium thiosulfate 100-500 g / L, potassium chloride 1-10 g / L, disodium hydrogen phosphate 3-12 g / L, appropriate amount of phosphoric acid, appropriate amount of sodium hydroxide; preferably, sodium thiosulfate 250 g / L, potassium chloride 4.4 g / L, disodium hydrogen phosphate 7.5-10 g / L, appropriate amount of phosphoric acid, appropriate amount of sodium hydroxide.

7. The sodium thiosulfate injection according to any one of claims 1 to 6, characterized in that: The pH value of the sodium thiosulfate injection is 7.5-9.5, preferably, the pH value is 8.0-8.

5.

8. The sodium thiosulfate injection according to any one of claims 1 to 7, characterized in that: The solvent of the sodium thiosulfate injection is distilled water, sterile water or water for injection, preferably water for injection.

9. The method for preparing the sodium thiosulfate injection according to any one of claims 1 to 8, characterized in that: The following steps are involved: (1) Add water for injection into the liquid preparation tank, and blow inert gas into the liquid preparation tank to control dissolved oxygen; (2) Add the prescribed amount of potassium chloride and disodium hydrogen phosphate and stir to dissolve, adjust the pH value to 8.5-9.0 with a pH adjuster such as phosphoric acid or sodium hydroxide, and then add sodium thiosulfate to dissolve; (3) pH adjuster: adjust the pH value to 7.5-9.0 with phosphoric acid or sodium hydroxide, and add a small amount of water for injection to make up the volume; (4) The drug solution is filtered through a filter element, filled, sealed, and sterilized by high-pressure wet heat to obtain a sodium thiosulfate injection.

10. The sodium thiosulfate injection according to any one of claims 1 to 8 or the sodium thiosulfate injection prepared by the preparation method according to claim 9 is used to treat or prevent ototoxicity, cyanide poisoning, eye diseases, hearing loss, urticaria, skin itching and other diseases in children or adults, or is used as calciphylaxis and disinfectant.

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