Azithromycin preparation and preparation method thereof

Abstract

The invention discloses an azithromycin preparation and a preparation method thereof. The azithromycin preparation comprises active pharmaceutical ingredients, a buffering agent and water, the active pharmaceutical ingredient is selected from at least one of azithromycin, azithromycin hydrate, pharmaceutically acceptable salts and isomers of azithromycin, and other substances capable of being converted into azithromycin and substances capable of releasing azithromycin in vivo; in the azithromycin preparation, the content of impurities is 0-1.2 wt%. Compared with oral azithromycin, the azithromycin composition has the advantages of quick effect taking, high bioavailability and the like, and compared with the existing preparation, the azithromycin composition is low in impurity content and low in dosage and can meet the clinical requirements of children on medication, new indications are added through non-clinical safety evaluation research and children safety and effectiveness clinical test research, and the application prospect is wide. The use method, dosage and clinical test data of children are provided in clinical use, the clarity and color of the solution meet the requirements, and the medication safety of the children can be better guaranteed.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical chemistry, and specifically relates to an azithromycin preparation and a preparation method thereof. Background Art

[0002] Azithromycin is a macrolide antibiotic and can be used for various infections such as skin infections, respiratory tract infections, and urogenital system infections caused by bacteria such as Chlamydia pneumoniae, Haemophilus influenzae, Legionella pneumophila, Moraxella catarrhalis, Staphylococcus aureus, or Streptococcus pneumoniae, and is one of the commonly used antibacterial drugs in clinical practice. Azithromycin has a broad antibacterial effect. The commercially available azithromycin dosage forms are mainly tablets and powders, and the injection is mainly an intravenous infusion, usually once a day.

[0003] Currently, the commercially available azithromycin for injection has problems such as a large number of impurities and a large total amount of impurities, unqualified inspection of solution clarity and color, too loose quality standard control, lack of pediatric medication guidance in clinical use, and single specification (the currently listed drug catalogs are all 0.5g specifications). Therefore, developing a new azithromycin to meet the clinical needs of pediatric medication has become a technical problem that needs to be solved urgently. Summary of the Invention

[0004] In order to solve the above problems, the present invention provides an azithromycin preparation and a preparation method thereof, which have the advantages of rapid onset and high bioavailability compared with oral azithromycin, and have low impurity content compared with the existing preparations, and low doses can meet the clinical needs of pediatric medication.

[0005] In order to achieve the above object, according to one aspect of the present invention, there is provided an azithromycin preparation, comprising a pharmaceutically active ingredient, a buffer, and water; the pharmaceutically active ingredient is selected from at least one of azithromycin, azithromycin hydrate, pharmaceutically acceptable salts of azithromycin, isomers, other substances that can be converted into azithromycin, and substances that can release azithromycin in vivo; in the azithromycin preparation, the impurity content is 0 to 1.2 wt%.

[0006] Preferably, the pharmaceutically active ingredient is azithromycin.

[0007] Among them, azithromycin hydrate includes monohydrate, dihydrate, etc., and anhydrous substance;

[0008] Pharmaceutically acceptable salts of azithromycin include citrate, fumarate, maleate, lactobionate, hydrochloride, sulfate, aspartate, malate, and tartrate.

[0009] In one embodiment of the present invention, in the azithromycin preparation, the content of the pharmaceutically active ingredient is 40 wt% to 50 wt%.

[0010] For example, 40 wt%, 41 wt%, 42 wt%, 43 wt%, 44 wt%, 45 wt%, 46 wt%, 47 wt%, 48 wt%, 49 wt%, 50 wt% or any point value within any of the above numerical ranges.

[0011] In one embodiment of the present invention, in the azithromycin preparation, the content of the buffer is 2.5 - 80 wt%; preferably 50 - 60 wt%, for example 55%.

[0012] In one embodiment of the present invention, the buffer is at least one group selected from the group consisting of a mixture of citric acid and sodium hydroxide, a mixture of citric acid and potassium hydroxide, a mixture of tartaric acid and sodium hydroxide, a mixture of malic acid and sodium hydroxide, and a mixture of lactic acid and sodium hydroxide.

[0013] Preferably, the buffer is a mixture of citric acid and sodium hydroxide, the content of citric acid is 3.8 - 76 wt%; the content of sodium hydroxide is 1.4 - 50 wt%, and the balance is water.

[0014] In one embodiment of the present invention, the preparation is an injection, preferably a sterile injection.

[0015] In one embodiment of the present invention, the water is water for injection.

[0016] In one embodiment of the present invention, the water content in the azithromycin for injection does not exceed 2.0%.

[0017] In one embodiment of the present invention, the azithromycin preparation further contains a cosolvent, and the cosolvent is selected from one or more of sulfuric acid, phosphoric acid, acetic acid, malic acid, maleic acid, acidic amino acids, citric acid, tartaric acid, fumaric acid, sodium dihydrogen phosphate, hydrochloric acid, and lactic acid, preferably citric acid.

[0018] In one embodiment of the present invention, the azithromycin preparation further contains a pH adjuster, and the pH adjuster is an acid or a base, for example selected from one or more of basic amino acids, organic amines, disodium hydrogen phosphate, sodium hydroxide, potassium hydroxide, citrates, and tartrates, preferably a 10 wt% sodium hydroxide solution.

[0019] In one embodiment of the present invention, the pH value of the azithromycin preparation is 6.2 - 6.8.

[0020] In one embodiment of the present invention, the azithromycin preparation is an injection, and the azithromycin for injection is a colorless and clear solution.

[0021] In one embodiment of the present invention, the total number of aerobic bacteria in every 100 ml of the azithromycin preparation does not exceed 10 cfu.

[0022] In one embodiment of the present invention, the endotoxin content in every 1 mg of azithromycin is less than 0.30 EU.

[0023] In one embodiment of the present invention, calculated according to the volume of the azithromycin injection, in terms of azithromycin, the concentration of the drug active ingredient is 90 - 110 mg / ml.

[0024] In one embodiment of the present invention, the density of the 0.2 g solution of azithromycin for injection is 1.070 g / ml.

[0025] In one embodiment of the present invention, the specification of the azithromycin for injection is 0.2 g, wherein there is 210 mg of azithromycin, 176.76 mg of citric acid, 73.5 mg of sodium hydroxide, an appropriate amount of 10 wt% sodium hydroxide; the injection water is added to 2.10 ml.

[0026] In one embodiment of the present invention, the particle size of the drug active ingredient: D 50 is 10 μm - 200 μm, D 90 is 50 μm - 350 μm.

[0027] In one embodiment of the present invention, in terms of the azithromycin preparation, according to the body weight of the administration object, when the body weight < 50 kg, it is administered at 10 mg / kg; when the body weight ≥ 50 kg, it is administered at 500 mg once a day.

[0028] In one embodiment of the present invention, the administration method of the azithromycin for injection can be subcutaneous injection, intradermal injection, intramuscular injection, intravenous injection. For example, it is administered by intramuscular injection.

[0029] The present invention also provides a preparation method of an azithromycin preparation for children, including steps of raw material weighing, liquid medicine preparation, sterilization filtration, filling, semi-capping, freeze-drying and capping; the liquid medicine preparation process includes:

[0030] S1, add injection water into the container, put the prescribed amount of citric acid into the container, stir to completely dissolve it, and then add the prescribed amount of sodium hydroxide, stir to completely dissolve it;

[0031] S2, slowly put the azithromycin raw material into the container, stir to completely dissolve it;

[0032] S3, prepare a pH value regulator, and adjust the pH value of the solution prepared in step S2 to make the pH value 6.2 - 6.8, to obtain the liquid medicine of the azithromycin preparation for children.

[0033] In one embodiment of the present invention, the pH value regulator is a 0.5 - 56% sodium hydroxide solution, preferably a 10 wt% sodium hydroxide solution.

[0034] The steps for preparing the pH regulator include: weighing an appropriate amount of sodium hydroxide, adding water for injection at 40 - 50°C to make up the volume, stirring to dissolve it to a concentration of 10 wt%, cooling it to below 35°C, and reserving it as the pH regulator.

[0035] For example, weigh 100.0 g of sodium hydroxide, add water for injection at 40 - 50°C to make up the volume to 1000 ml, stir to dissolve it, and cool it to below 35°C, then reserve it as a 10 wt% sodium hydroxide solution.

[0036] In an embodiment of the present invention, magnetic stirring is used during stirring in the feeding process, and the frequency of the magnetic pump is maintained at 90% - 100%, preferably 92% - 98%.

[0037] In an embodiment of the present invention, in step S3, each time when adding the 10 wt% sodium hydroxide solution to adjust the pH value, sample is taken for detection after magnetic stirring for 10 minutes.

[0038] In an embodiment of the present invention, before preparing the liquid medicine, it also includes: checking the endotoxin content of the water for injection to meet the water for injection standard.

[0039] In an embodiment of the present invention, it also includes: before feeding, first purge the interlayer of the container to drain the water in the interlayer; during the addition of sodium hydroxide, cool water is introduced into the interlayer of the container to cool the solution in the container to below 25°C.

[0040] In the sterilizing filtration step, pre - filtration and terminal sterilizing filtration are adopted. For example, a 0.45 μm pre - filter with a filter element made of polyethersulfone, a 0.22 μm redundant filter, and a 0.22 μm terminal sterilizing filter are connected in series, and the sterilizing operation is carried out by controlling the sterilizing pressure ≤ 1.5 bar.

[0041] In an embodiment of the present invention, the time interval from the start of liquid medicine preparation to the start of sterilizing filtration shall not exceed 8 hours; the time from the start of sterilizing filtration to the end of sterilizing filtration of the liquid medicine shall not exceed 5 hours.

[0042] In an embodiment of the present invention, in the filling step, according to the single - dose filling amount required for clinical use, add water for injection below 30°C to make up the volume. During volume - making, the magnetic pump stops stirring, and after volume - making is completed, the magnetic pump is started to stir. The frequency of the magnetic pump is 95%, and stirring is carried out for 15 minutes. The temperature of the solution after volume - making is below 30°C; sample is taken to detect the intermediate, and after sampling is completed, the magnetic pump is closed and the liquid medicine is allowed to stand.

[0043] The beneficial effects of the present invention:

[0044] The present invention provides an azithromycin injection, which has the advantages of rapid onset and high bioavailability compared with oral azithromycin. Compared with the commercially available azithromycin for injection, the present invention controls the impurity content of the product by using high-quality raw materials, formulating strict quality standards for the bulk drug, scientific formulation prescriptions, optimal production process parameters, well-compatible inner packaging materials, and the strictest formulation quality standards. The impurity limit is strictly controlled, the quality controllability is strong, and the safety is high. Three specifications (0.1 g, 0.2 g, 0.3 g) of azithromycin for injection have been developed. Through non-clinical safety evaluation studies, clinical trials on the safety and efficacy of children, new indications have been added, and the usage and dosage for children and clinical trial data have been provided in clinical use. The clarity and color of the solution meet the requirements, which can better ensure the safety of children's medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 shows the particle size and particle size distribution of azithromycin in Example 1 Figure 1 ;

[0046] Figure 2 shows the particle size and particle size distribution of azithromycin in Example 1 Figure 2 . DETAILED DESCRIPTION OF THE EMBODIMENTS

[0047] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below with reference to the embodiments. It should be emphasized that the specific embodiments described here are only used to better illustrate the present invention, and are partial embodiments of the present invention, not all embodiments, so they are not used to limit the present invention. In addition, the technical features involved in the embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0048] The term "injection" refers to an injectable composition (usually subcutaneous injection or intramuscular injection), and after injection, the drug is slowly distributed into the systemic circulation system of the subject being used. In this way, the drug can be continuously delivered in a controlled manner.

[0049] Example 1

[0050] The production process (taking 40,000 vials per batch as an example) is described as follows:

[0051] 1 Inner packaging material treatment

[0052] 1.1 Treatment of controlled injection vials

[0053] (1) Bottle sorting

[0054] Remove the outer packaging of the controlled injection vials. After wiping and disinfecting the inner packaging, they enter the vial and stopper storage room. Invert the packaging boxes twice to remove debris. When arranging the vials, pick out any unqualified vials (such as those with a dropped bottom, cracked opening, crooked neck, thick neck, thin neck, nodules, etc.) at any time. Place the trays of controlled injection vials on a special trolley in the D-class clean area and transfer them through the air lock chamber to the vial washing room. Before washing, distribute them on the conveyor belt of the vial conveyor.

[0055] (2) Vial washing

[0056] ① Parameter setting: The circulating water temperature is 50 - 60 °C, the number of workstations for circulating water flushing is 2, the circulating water pressure ≥ 0.2 MPa, the number of workstations for compressed air is 1, the compressed air pressure ≥ 0.3 MPa, the number of workstations for injection water flushing is 1, the injection water pressure ≥ 0.2 MPa, the number of workstations for compressed air is 2, the compressed air pressure ≥ 0.3 MPa, and the ultrasonic wave is set at 300 - 500 W.

[0057] ② Vial washing process: Place the controlled injection vials on the small conveyor belt of the vial washer. After ultrasonic treatment, they go through the following steps: circulating water flushing of the inner wall → circulating water flushing of the outer wall → circulating water flushing of the inner wall → discharging the water in the vial with compressed air → injection water flushing of the inner wall (through a 0.22 μm membrane) → discharging the water in the vial with compressed air → discharging the water in the vial with compressed air → compressed air flushing of the outer wall, and then enter the tunnel oven for sterilization.

[0058] (3) Vial drying

[0059] ① Parameter setting: The sterilization temperature of the sterilization and drying machine is 320 °C. Turn the conveyor belt knob to adjust the conveyor belt frequency converter to display ≤ 43.5 HZ. Set the conveyor belt to automatic. After the temperature rises to 320 ± 10 °C, the conveyor belt starts to move the vials. Adjust the vial washing speed of the vial washer to the 24 - 27 knob position. The pressure difference between the preheating section and the room is between 2 and 12 Pa, the pressure difference between the heating section and the room is between 2 and 12 Pa, and the pressure difference between the cooling section and the room is between 2 and 12 Pa. The pressure difference between the heating section and the room should be greater than the pressure differences between the cooling section and the preheating section and the room. Check and record the temperature displayed on the touch screen of the sterilization and drying machine every 30 minutes. When the controlled injection vials are washed to the air damper at the preheating section and the heating section, the air damper can be lifted to the specified position.

[0060] ② Vial drying process: After the washed and sterilized controlled injection vials are cooled in the cooling area of the sterilization and drying machine, they enter the filling room.

[0061] ③ Rear empty operation: When the controlled injection vials move to the inside of the heating section and are close to the cooling section, manually pause the conveyor belt and time for 5 minutes, then start the conveyor belt again to continue the vial washing operation.

[0062] ④ Front empty operation: After all the washed controlled injection vials leave the preheating section and enter the heating section through the conveyor belt, manually pause the conveyor belt (stop filling) and time for 5 minutes, then start the conveyor belt again to continue the filling operation.

[0063] ⑤ Precautions: Sterilize the injection vials within 2 hours after cleaning; the sterilized and clean injection vials shall not be used more than 4 hours after the sterilization is completed (stored under Class A conditions). If the time limit is exceeded, the sterilized vials shall be treated as waste.

[0064] ⑥ Use 4ml neutral borosilicate glass control injection vials.

[0065] 1.2 Stopper treatment

[0066] Table 1. Setting table of stopper process parameters

[0067]

[0068] (1) Stopper cleaning and sterilization: Use a stopper cleaning machine. First, suck the stoppers to be washed into the machine body by vacuum suction, and then operate according to the cleaning and sterilization procedure. After the fine cleaning is completed, QA samples to check the visible foreign matters in the cleaning water. After passing the inspection, perform silicification, rinsing, and then transfer to the sterilization procedure. After the sterilization is completed, send the air from the fan into the Class A clean air. After the chamber of the stopper cleaning machine returns to normal pressure, take out the stoppers from the stopper outlet of the filling chamber. The stopper taking-out is carried out under the Class A air supply.

[0069] (2) Precautions: The time from stopper cleaning to sterilization shall not exceed 4 hours. If the time limit is exceeded, re-cleaning is required. After the stopper sterilization is completed, the stoppers shall be stored in the stopper cleaning machine for no more than 25 hours and transferred to the Class A laminar flow storage for no more than 9 hours. The used stoppers cannot be recycled and reused, and the sterilized but unused stoppers cannot be sterilized and reused either. They shall all be treated as waste.

[0070] (3) Add dimethicone to the 13G stoppers at a ratio of 1 - 2 ml per 10,000 vials.

[0071] 1.3 Aluminum cap treatment

[0072] Table 2. Setting table of aluminum cap cleaning machine process parameters

[0073]

[0074] (1) Aluminum cap cleaning and sterilization: Use a fully automatic ultrasonic aluminum cap cleaning machine for positioning and feeding, and operate according to the cleaning and sterilization procedure. After the hot air drying is completed, the aluminum cap machine collects the Class A clean air for intake and recompression. After the vacuum in the aluminum cap machine returns to normal pressure and the temperature in the box is less than 70 °C, discharge the products under the protection of the Class A air supply.

[0075] (2) Precautions: After the aluminum cap sterilization is completed, the aluminum caps shall be stored in the aluminum cap cleaning machine for no more than 25 hours and transferred to the Class A laminar flow storage for no more than 9 hours.

[0076] (3) Use Φ13 aluminum caps.

[0077] 2 Weighing

[0078] Turn on the dust removal unit, turn on the power switch of the negative pressure weighing unit, turn on the fan switch, confirm whether the fan parameters meet the requirements, let the weighing unit self-purify for more than 10 minutes, and preheat the electronic balance.

[0079] Before weighing, check the name, incoming batch number, quantity, etc. of the raw and auxiliary materials. During weighing, one person should weigh and one person should recheck to ensure accuracy. QA independently rechecks to ensure that the weighed raw and auxiliary materials meet the requirements of the production order.

[0080] After weighing the materials each time, write the material label (information such as name, quantity, etc.) and paste it on the outside of the material weighing bag. After weighing is completed, the negative pressure weighing unit should self-purify for more than 10 minutes before it can be turned off. Do a good job in cleaning and clearing the operation room and end the work.

[0081] According to the batch production order form of 0.2g (40,000 pieces) of azithromycin for injection, turn on the dust removal unit and weigh the specified quantity of azithromycin, citric acid, sodium hydroxide, etc. in the negative pressure weighing unit of the weighing room.

[0082] 3 Preparation of liquid medicine

[0083] (1) Detection of preparation water

[0084] Take the injection water in the dilution tank to check the endotoxin, which is required to meet the standard of injection water.

[0085] (2) Preparation of liquid medicine

[0086] Preparation method of 10wt% sodium hydroxide solution: Weigh 100.0g of sodium hydroxide and make up the volume to 1000ml with injection water at 40 - 50°C, stir to dissolve, cool to below 35°C, and use it as 10wt% sodium hydroxide solution for standby. Prepare 2000ml of 10wt% sodium hydroxide solution according to production needs.

[0087] In the dilution room, add 58.80kg of injection water to the dilution tank, with the water temperature not higher than 35°C. Put the weighed prescription amount of citric acid (7070.4g) into the dilution tank and stir for 10 minutes to completely dissolve it. During stirring, the frequency of the magnetic pump is 95% (during the feeding period, the magnetic pump has been running at a frequency of 95%). Then add the prescription amount of sodium hydroxide (2940.0g) and stir for 10 minutes to completely dissolve it. During the addition of sodium hydroxide, cool water is introduced into the jacket of the dilution tank to cool the solution to below 25°C. After the solution temperature meets the requirements, drain the cooling water in the jacket. (Before feeding, first purge the jacket to drain all the water that may exist in the jacket. During the process, record the weight of the solution after dissolving citric acid completely. Adding the weight of the prescription amount of sodium hydroxide is the weight of the solution before adding cooling water. After draining the cooling water, purge the jacket with compressed air to this weight, which is regarded as meeting the requirement of emptying the jacket).

[0088] Slowly put 8400.0 g of azithromycin raw material (dry and pure, purchased from Shijiazhuang Pharmaceutical Group Ouyi Pharmaceutical Co., Ltd.) into the dilute preparation tank, stir until it is completely dissolved, and stir for another 20 minutes after visually checking that there is no visible matter on the liquid surface. Take a sample to measure the pH value of the solution, and adjust the pH value to 6.2 - 6.8 with 10 wt% sodium hydroxide solution (each time 10 wt% sodium hydroxide solution is added for adjustment, stir for 10 minutes and then take a sample for detection).

[0089] The density of the 0.2 g solution of azithromycin for injection is 1.070 g / ml, the batch size is 40,000 vials, the single vial filling volume is 2.1 ml according to clinical use requirements, and make up the volume to 89.88 kg with injection water below 30°C (the magnetic pump stops stirring during volume makeup, and starts stirring after volume makeup is completed). Stir for 15 minutes, the frequency of the magnetic pump during stirring is 95%, and the temperature of the solution after volume makeup is below 30°C. Take a sample and detect the intermediate product. After sampling is completed, turn off the magnetic pump and let the liquid medicine stand still.

[0090] Table 3. Key process parameters of the preparation post

[0091]

[0092] Table 4. Quality standards of intermediate products

[0093]

[0094] 4 Sterilization filtration

[0095] (1) Use a 0.45 μm pre-filter, a 0.22 μm sterilization filter (redundant filter) and a 0.22 μm terminal sterilization filter (main filter) with a polyethersulfone filter element material in series, and control the sterilization pressure to be ≤ 1.5 bar for sterilization operation.

[0096] (2) The integrity needs to be checked before the first use and after the 10th use of the redundant filter. The filter element of the main filter needs to be checked for integrity before and after use. If the integrity test fails, the filter element of the redundant filter needs to be checked for integrity again. If the integrity test passes, the product can still be released.

[0097] (3) Before each integrity test, the filter should be wetted with injection water. The integrity test result of the 0.22 μm filter element is ≥ 3320 mbar to be qualified;

[0098] (4) The time interval from the start of preparation to the start of sterilization filtration of the liquid medicine shall not exceed 8 hours. The time from the start of sterilization filtration to the end of sterilization filtration shall not exceed 5 hours. Filling can start after the intermediate product inspection (except for the microbial limit item) is qualified.

[0099] Table 5. Information table of plastic components

[0100]

[0101] 5 Filling and semi-stoppering

[0102] (1) Preparation, treatment and assembly of filling accessories and liquid receiving tank

[0103] Close the manual valve at the bottom of the liquid receiving tank, disassemble the heat exhaust pipe head connected to the manual valve of the liquid receiving tank, and connect the hose connecting the distributor to the manual valve of the liquid receiving tank in a Class A environment. After assembling the filling components, connect them to the distributor, and then open the manual valve at the bottom of the liquid receiving tank to allow the liquid medicine to enter the filling device through the liquid receiving tank. Dock the LCE laminar flow cart with the lower tray of the filling machine; prepare to start filling.

[0104] (2) Filling process

[0105] ① Adjust the position of the needle so that the needle is aligned with the center of the mouth of the controlled injection vial. Adjust the filling volume. Each needle is adjusted 2 - 3 times, and the filling volume is adjusted to 2.1 ml (2.247 g). Calculate the filling volume according to the intermediate product, and control the filling volume difference within the range of ±4%. The first 4 sub-packagings of the intermediate product for each needle are all treated as waste. Control the filling speed at 260 - 300 vials per minute, and monitor the filling volume every 30 minutes. Control the semi-stoppering height within the range of 3 - 5 mm.

[0106] ② After filling and semi-stoppering, directly enter the LCE laminar flow cart at the lower tray. After it is full, replace it with another LCE laminar flow cart for filling. Dock the laminar flow cart filled with the intermediate product with the shelf of the freeze dryer, and send the intermediate product into the partition of the freeze dryer through the small door of the freeze dryer. After the filling is completed, insert the freeze-drying temperature probe, close the small door of the freeze dryer, and notify the freeze-drying post to start freeze-drying.

[0107] (3) Time limit control: The time interval from the start of aseptic filtration to the end of filling shall not exceed 7 hours. The time from the end of filling to the start of freeze-drying shall not exceed 1 hour.

[0108] Note: If the time limit is exceeded, the liquid medicine, rubber stoppers, and injection vials shall all be treated as waste. They shall not be recycled for the production of preparations.

[0109] 6 Freeze-drying

[0110] 6.1 Preparation before starting the GLZY33BS vacuum freeze dryer

[0111] Before starting the machine, check whether the circulating water pressure, voltage, compressed air, compressor, and lubricating oil of the vacuum pump are within the specified range. Check whether the power supply and valves at all places are in normal state. Start the machine, enter the "freeze-drying" program, input the correct drug name and batch number, and click on manual operation. Requirements: Cooling circulating water pressure ≥ 0.1 MPa; Compressed air pressure ≥ 0.2 MPa.

[0112] 6.2 Freeze-drying process

[0113] When the product is put into the box, control the shelf temperature at 20°C or below.

[0114] (1) Pre-freezing:

[0115] Turn on the circulation pump. Turn on the refrigeration compressors 1 and 2 to collect gas. When the low-pressure gauge indicates below "0", open the plate cooling valves 1 and 2, and at the same time click to enter the "Trend Curve" program and click Start (must click), and the freeze-drying program starts recording. Control the shelf temperature to -45°C (temperature fluctuation range -45±5°C, the product temperature reaches below -35°C, freeze for more than 3 hours, and then sublime.

[0116] (2) Sublimation drying

[0117] After pre-freezing reaches the required temperature for about 1 hour or more, close the plate cooling valves 1 and 2, start the rear box cooling valves 1 and 2 to refrigerate the condenser. When the condenser temperature reaches below -56°C, start sublimation. First, turn on the vacuum pumps 1 and 2 (for 20 seconds), open the small butterfly valve between the vacuum pump and the condenser (for 1 - 2 minutes), then open the middle partition valve between the condenser and the freeze-drying chamber (for 20 - 30 seconds). When the vacuum pressure in the front chamber reaches below 20 Pa, set the heat transfer oil temperature to rise to -8°C in 1 hour, turn on the electric heating, and the first sublimation starts. Keep at -8°C for 15 - 23 hours. The operator observes the water line every half hour and closely pays attention to the changes in the medicine, whether there are phenomena of stratification and agglomeration. The disappearance of the water line basically indicates the end of the first sublimation. After the medicine turns completely white, set the heat transfer oil temperature to rise from -8°C to 40 - 45°C within 6 - 10 hours for desorption drying.

[0118] Desorption drying:

[0119] When the product temperature reaches above 35°C, release the vacuum control and keep it at 40 - 45°C for more than 4 hours. When the curves of the shelf temperature and the product temperature tend to be consistent, the freeze-drying is completed.

[0120] (3) Plugging:

[0121] After the desorption drying is completed, turn on the hydraulic pump. First, turn off the electric heating and the middle partition valve, then turn off the small butterfly valve, turn off the Roots pump, turn off the vacuum pumps 1 and 2, turn off the cold traps 1 and 2, the screw chillers 1 and 2, turn off the compressors 1 and 2, and turn off the circulation pump. Fill high-purity nitrogen until the absolute pressure in the front chamber is 50 - 55 KPa and then plug. Turn on the hydraulic pump, click ↓, and finally the compensation plate layer should be pressed on the top of the plate layer, and close the freeze-drying curve.

[0122] (4) Unloading from the box:

[0123] Open the front box intake valve (for 10 - 20 minutes). When the vacuum in the front box returns to normal pressure, notify the filling personnel in the aseptic area to open the box door. After receiving the notice, the filling personnel in the aseptic area notify the capping personnel to prepare to receive the drugs for capping, open the box door, and take out the drugs from the box in the reverse order of each layer during boxing (from bottom to top, from outside to inside, layer by layer).

[0124] (5) Defrosting:

[0125] After taking out the drugs from the box, enter the defrosting procedure. First, open the rear box air release valve. After normal pressure, open the drain valve and overflow valve in sequence, then open the rear box water inlet valve, and defrost by immersion and spraying with purified water. After defrosting, first close the rear box water inlet valve. After the water in the rear box is drained, close the rear box overflow valve and drain valve.

[0126] 7 Capping

[0127] Before capping, wipe the capping machine with 75% ethanol or 70% isopropyl alcohol, then place the clean aluminum caps into the oscillating tray of the machine, turn on the capping machine, adjust the cap feeding speed: less than 400 vials per minute for 4ml injection vials, adjust the capping speed: 150 - 175 vials per minute for 4ml injection vials, start the capping machine for trial capping, and check whether the cap feeding and capping speeds match during capping, and whether there are phenomena such as skirt edges and wrinkling. During the capping process, on the basis of the three-finger method, check with a full-automatic torque tester, and the required torque should be greater than 0.126 N·M. After passing the debugging, carry out capping formally. During the capping process, check the capping quality every half hour, which should be tight, smooth, flat, and without wrinkles. Attach an identification card to the product after capping. The vials should be capped as soon as possible after being stoppered.

[0128] 8 Visual inspection

[0129] (1) Classify and place the products with loose capping, cracked bottles, damaged, agglomerated, incorrect filling volume, unqualified color, and foreign objects detected during visual inspection, and place obvious signs indicating the product name, specification, batch number, quantity, reason for non-conformance, etc. After the end of batch production, the team leader counts the quantity and collects and hands it over to the administrative department for processing in a timely manner.

[0130] (2) Place the qualified products after visual inspection in blue transfer boxes, fill each box neatly (400 vials per box for 4ml injection vials), and place the production status identification card, filling in the product name, specification, batch number, quantity, worker number, etc., and transport them to the transfer room or transfer station for packaging.

[0131] Azithromycin for injection prepared by the above method is a sterile preparation made from azithromycin with appropriate citric acid and sodium hydroxide as buffer agents, and is a white or off-white loose mass or powder. The content of azithromycin (C 38 H 72 N 2 O 12 ) should be 95.0% - 115.0% of the labeled amount.

[0132]

Identification

[0133] (1) Test according to the thin layer chromatography method (General Principles 0502, Section IV, Chinese Pharmacopoeia 2020 Edition).

[0134] Test solution: Take an appropriate amount of this product, dissolve and dilute with absolute ethanol to prepare a solution containing about 20 mg of azithromycin per 1 ml.

[0135] Reference solution: Take an appropriate amount of azithromycin reference substance, dissolve and dilute with absolute ethanol to prepare a solution containing about 20 mg of azithromycin per 1 ml.

[0136] Chromatographic conditions: Use a silica gel G thin layer plate, and use ethyl acetate - n - hexane - diethylamine (10:10:2) as the developing solvent.

[0137] Determination method: Pipette 5 μl each of the test solution and the reference solution, spot them on the same thin layer plate respectively, develop, air - dry, spray with the developer (take 2.5 g of sodium molybdate and 1 g of cerium sulfate, dissolve and dilute to 100 ml with 10% sulfuric acid solution), let stand for 10 minutes, then heat at 105 °C for 4 - 5 minutes.

[0138] Result judgment: The color and position of the main spot shown by the test solution should be the same as those of the main spot of the reference solution.

[0139] In the chromatogram recorded under the content determination item, the retention time of the main peak of the test solution should be consistent with that of the main peak of the reference solution.

[0140]

Inspection

[0141] Acidity: Take an appropriate amount of this product, add water to prepare a solution containing about 100 mg of azithromycin per 1 ml, and determine according to law (General Principles 0631, Section IV, Chinese Pharmacopoeia 2020 Edition). The pH value should be 6.2 - 6.8.

[0142] Clarity and color of solution: Take 5 bottles of this product, add water to prepare a solution containing 100 mg of azithromycin per 1 ml respectively, and immediately examine. The solution should be clear and colorless.

[0143] Related substances: Determine according to the high performance liquid chromatography method (General Principles 0512, Section IV, Chinese Pharmacopoeia 2020 Edition). Prepare freshly before use or use a low - temperature injector.

[0144] Diluent: Ammonium dihydrogen phosphate solution (weigh 1.73 g of ammonium dihydrogen phosphate, dissolve and dilute to 1000 ml with water, adjust the pH value to 10.0 with ammonia water) - methanol - acetonitrile (7:7:6).

[0145] Test solution: Weigh an appropriate amount of this product accurately, dissolve it in an appropriate amount of water (dissolve 75 mg of azithromycin in 1 ml of water), and quantitatively dilute it with the diluent to prepare a solution containing about 7.5 mg of azithromycin per 1 ml.

[0146] Reference solution: Accurately measure 1 ml of the test solution, transfer it to a 100-ml volumetric flask, dilute it to the mark with the diluent, and mix well.

[0147] Blank excipient solution: Weigh accurately about 126.0 mg of citric acid and about 52.5 mg of sodium hydroxide, place them in the same 20-ml volumetric flask, dissolve them in 2 ml of water, then dilute to the mark with the diluent, and mix well.

[0148] System suitability solution: Weigh about 7.5 mg of azithromycin reference substance, dissolve it in 1 ml of the diluent, and mix well.

[0149] Chromatographic conditions: Use octadecylsilane chemically bonded silica gel as the filler; use phosphate buffer solution (0.90 g / L disodium hydrogen phosphate anhydrous solution, adjust the pH to 8.9 with dilute phosphoric acid or dilute sodium hydroxide solution) as mobile phase A, and use methanol-acetonitrile (250:750) as mobile phase B, perform linear gradient elution according to Table 6 below; the column temperature is 60 °C; the flow rate is 1.0 ml per minute; the detection wavelength is 210 nm; the injection volume is 50 μl.

[0150] Table 6. Linear gradient elution parameters

[0151]

[0152] System suitability requirements: In the system suitability solution, the retention time of the azithromycin peak should be between 40 and 50 minutes, and the theoretical plate number of azithromycin should not be less than 3000.

[0153] Determination method: Accurately measure the test solution, reference solution and blank excipient solution, inject them into the liquid chromatograph respectively, and record the chromatogram.

[0154] Limit: If there are impurity peaks in the chromatogram of the test solution, after deducting the blank excipient peak, for impurity L (relative retention time is about 0.27), the calculated peak area after correction (multiplied by the correction factor 2.3) shall not be greater than 0.2 times (0.2%) of the main peak area of the control solution; for impurity F (relative retention time is about 0.49), the calculated peak area after correction (multiplied by the correction factor 0.3) shall not be greater than 0.2 times (0.2%) of the main peak area of the control solution; the sum of the peak areas of impurities D, J, and Q (relative retention time is about 0.53) shall not be greater than 0.3 times (0.3%) of the main peak area of the control solution; the peak area of impurity B (relative retention time is about 1.37) shall not be greater than 0.5 times (0.5%) of the main peak area of the control solution; the peak area of any other single impurity shall not be greater than 0.2 times (0.2%) of the main peak area of the control solution; the sum of the peak areas of each impurity calculated after correction shall not be greater than 1.2 times (1.2%) of the main peak area of the control solution. Peaks less than 0.05 times (0.05%) of the main peak area of the control solution shall be ignored.

[0155] Water: Take this product and determine it according to the method for the determination of water (General Chapter 0832, Method 1 in Part IV of the Chinese Pharmacopoeia 2020 Edition). The water content shall not exceed 2.0%.

[0156] Bacterial endotoxin: Take this product and examine it according to the law (General Chapter 1143 in Part IV of the Chinese Pharmacopoeia 2020 Edition). The amount of endotoxin contained in every 1 mg of azithromycin shall be less than 0.30 EU.

[0157] Sterility: Take this product, dissolve and dilute it with 0.9% sterile sodium chloride to make a solution containing 20 mg of azithromycin per 1 ml. After treatment by the membrane filtration method, wash it with 0.1% sterile peptone aqueous solution in several portions (not less than 600 ml per membrane). Using Staphylococcus aureus as the positive control bacterium, examine it according to the sterility test method (General Chapter 1101 in Part IV of the Chinese Pharmacopoeia 2020 Edition), and it shall meet the requirements.

[0158] Others: It shall comply with the relevant regulations under the injection dosage form (General Chapter 0102 in Part IV of the Chinese Pharmacopoeia 2020 Edition).

[0159]

Assay

[0160] Determine according to the high performance liquid chromatography method (General Chapter 0512 in Part IV of the Chinese Pharmacopoeia 2020 Edition).

[0161] Diluent: Phosphate buffer solution (0.04 mol / L dipotassium hydrogen phosphate solution, adjusted to pH 8.2 with 20% phosphoric acid)-acetonitrile (40:60).

[0162] Test solution: Take an appropriate amount of the content under the difference in filling quantity, weigh it accurately, dissolve it with the diluent and quantitatively dilute it to make a solution containing about 0.5 mg per 1 ml.

[0163] Reference substance solution: Weigh an appropriate amount of azithromycin reference substance accurately, dissolve it in the diluent and quantitatively dilute it to prepare a solution containing about 0.5 mg per 1 ml.

[0164] Chromatographic conditions: Use octadecyl-bonded polyvinyl alcohol as the filler; use phosphate buffer solution (take 0.04 mol / L dipotassium hydrogen phosphate solution, adjust the pH value to 11.0 with 5 mol / L potassium hydroxide solution)-acetonitrile (40:60) as the mobile phase; the column temperature is 40 °C; the detection wavelength is 210 nm; the injection volume is 10 μl.

[0165] System suitability requirements: The number of theoretical plates calculated based on the azithromycin peak shall not be less than 3000, and the tailing factor shall be less than 1.5.

[0166] Assay method: Accurately measure the test solution and the reference substance solution, inject them into the liquid chromatograph respectively, and record the chromatogram. Calculate according to the external standard method with the peak area.

[0167]

Determination of particle size and particle size distribution

[0168] Use the azithromycin in Example 1 above to determine the particle size and particle size distribution with a laser particle size analyzer. The results are shown in Figure 1 and Figure 2 . It can be seen from Figure 2 that the particle size D 50 of the azithromycin sample is 10 μm - 200 μm, and D 90 is 50 μm - 350 μm, and the particle size distribution is relatively wide.

[0169] This product is a freeze-dried powder for injection, and the particle size of the raw material does not affect the final quality of the preparation.

[0170]

Microscopic imaging of particles

[0171] Use the azithromycin for injection prepared in Example 1 above, dilute the azithromycin for injection with injection water, and observe its morphology and aggregation with a microscope. The particle size of this sample is consistent with the results of laser particle size determination, indicating that the prepared azithromycin preparation is relatively stable and has uniform particle size.

[0172] Test 1

[0173]

Animal toxicity test

[0174] Perform a toxicity test by repeatedly intravenously infusing the azithromycin for injection obtained in the present invention to juvenile Beagle dogs for 4 weeks and a recovery period of 4 weeks.

[0175] Thirty-two Beagle dogs aged 9 to 11 weeks were used in the experiment. They were randomly divided into 4 groups, namely the low-dose, medium-dose, high-dose groups of the test article and the negative control group. The animals in each group were given azithromycin for injection and normal saline by intravenous infusion respectively, and the administration was repeated for 28 days.

[0176] During the experiment, no death or near-death phenomenon occurred in the animals of each group. No abnormality was observed in the local injection site of each group of animals. Some individual animals showed single vomiting symptoms, and loose stools, bloody stools and other symptoms appeared after drug withdrawal, which were mainly considered as gastrointestinal reactions caused by azithromycin; fatty degeneration occurred in the splenic small somatic cells of some animals. Under the conditions of this experiment, the main toxic reaction was fatty degeneration in the splenic small somatic cells.

[0177] Experiment 2

[0178]

Active systemic anaphylaxis test in animals

[0179] Twenty-four male guinea pigs were used for sensitization in the experiment. They were randomly divided into 4 groups according to body weight segments, with 6 in each group, namely the negative control group, positive control group, low-dose group of the test article, and high-dose group.

[0180] During sensitization, intraperitoneal injection was used for administration, once every other day, for a total of 3 times. On the 14th day after the last sensitization, intravenous injection was given in the foot for challenge, and the allergic reaction symptoms appeared in the animals of each group were observed after challenge. Under the conditions of this experiment, no allergic reaction symptoms were seen in the guinea pigs, and the allergic reaction was negative.

[0181] Experiment 3

[0182]

In vitro hemolysis test

[0183] The azithromycin for injection preparation prepared above according to the present invention was used to observe whether hemolysis or aggregation of rabbit red blood cells was caused in vitro, so as to provide in vitro test data for clinical research.

[0184] Under the conditions of this experiment, azithromycin for injection at a concentration of 1 mg / ml had no hemolytic effect on rabbit red blood cells in vitro and did not cause red blood cell aggregation.

[0185] Experiment 4

[0186]

Local irritation test research

[0187] Combined with the local observation during animal toxicity testing, at the end of the last administration and the recovery period, subcutaneous bleeding at the injection site to varying degrees was seen in some animals, which was considered as mechanical injury caused by drug administration or anesthesia injection, and no irritant change of the test article was seen.

[0188] Experiment 5

[0189] Compared with rapid intravenous administration, the change in plasma drug concentration during intravenous administration is closer to that of oral administration in clinical applications. The reproductive and developmental toxicity of intravenous administration can be evaluated by oral administration tests. Therefore, the reproductive and developmental toxicity test of intravenous administration was not conducted.

[0190] Reproductive toxicity tests in rats and mice have shown that when the dosage of azithromycin reaches the level of a moderately toxic maternal dose (i.e., 200 mg / kg / day, calculated by body surface area mg / m 2 , which is about 2 to 4 times the human drug dosage of 500 mg / kg / day), no teratogenic effects were found, and no damage to fertility and fetuses was found.

[0191] The above are only the preferred application embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the technical principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. An azithromycin preparation, characterized in that, the azithromycin preparation contains a drug active ingredient, a buffer and water, and the drug active ingredient is selected from at least one of azithromycin, azithromycin hydrate, a pharmaceutically acceptable salt of azithromycin, an isomer, other substances capable of being converted into azithromycin, and substances capable of releasing azithromycin in vivo; in the azithromycin preparation, the impurity content is 0 to 1.2 wt%.

2. The azithromycin preparation according to claim 1, characterized in that, the preparation is an injection, preferably a sterile injection. Preferably, the drug active ingredient is azithromycin. Preferably, in the azithromycin preparation, the content of the drug active ingredient is 40 wt% to 50 wt%. Preferably, in the azithromycin preparation, the content of the buffer is 2.5 to 80 wt%; preferably 50 to 60 wt%. Preferably, the buffer is at least one group of a mixture of citric acid and sodium hydroxide, a mixture of citric acid and potassium hydroxide, a mixture of tartaric acid and sodium hydroxide, a mixture of malic acid and sodium hydroxide, and a mixture of lactic acid and sodium hydroxide. Preferably, the buffer is a mixture of citric acid and sodium hydroxide, the content of citric acid is 3.8 to 76 wt%; the content of sodium hydroxide is 1.4 to 50 wt%, and the balance is water.

3. The azithromycin preparation according to claim 1, characterized in that, the azithromycin preparation further contains a cosolvent, and the cosolvent is selected from one or more of sulfuric acid, phosphoric acid, acetic acid, malic acid, maleic acid, acidic amino acids, citric acid, tartaric acid, fumaric acid, sodium dihydrogen phosphate, hydrochloric acid, and lactic acid. Preferably citric acid.

4. The azithromycin preparation according to claim 1, characterized in that, the azithromycin preparation further contains a pH regulator, and the pH regulator is selected from one or more of basic amino acids, organic amines, disodium hydrogen phosphate, sodium hydroxide, potassium hydroxide, citrate salts, and tartrate salts. Preferably a 10 wt% sodium hydroxide solution. Preferably, the water content in the injectable azithromycin does not exceed 2.0 wt%, and the pH value is 6.2 to 6.

8. Preferably, the total number of aerobic bacteria in every 100 ml of the test sample does not exceed 10 cfu. Preferably, the amount of endotoxin contained in every 1 mg of injectable azithromycin is less than 0.30 EU.

5. The azithromycin preparation according to claim 1, characterized in that, calculated according to the volume of the injection, based on azithromycin, the concentration of the drug active ingredient is 90 to 110 mg / ml. Preferably, the density of the 0.2 g injectable azithromycin solution is 1.070 g / ml. For example, the specification of the injectable azithromycin is 0.2 g, wherein, azithromycin is 210 mg, citric acid is 176.76 mg, sodium hydroxide is 73.5 mg, 10% sodium hydroxide is appropriate; water for injection is added to 2.10 ml.

6. The azithromycin preparation according to claim 1, characterized in that, The particle size characteristics of the drug active ingredient in the azithromycin preparation are as follows: D 50 is 10 μm - 200 μm, D 90 is 50 μm - 350 μm. Preferably, based on the azithromycin preparation, according to the body weight of the administration subject, if the body weight < 50 kg, administer at 10 mg / kg; if the body weight ≥ 50 kg, administer 500 mg once a day.

7. A method for preparing the azithromycin preparation according to any one of claims 1 to 6, characterized in that it includes steps of raw material weighing, liquid medicine preparation, sterilization filtration, filling, semi-capping, freeze-drying and capping; the liquid medicine preparation includes: S1, add water for injection into the container, put the prescribed amount of citric acid into the container, stir to completely dissolve it, then add the prescribed amount of sodium hydroxide and stir to completely dissolve it; S2, slowly put the azithromycin raw material into the container and stir to completely dissolve it; S3, prepare a pH regulator and adjust the pH value of the solution prepared in step S2 to make the pH value 6.2 - 6.8 to obtain the azithromycin preparation liquid medicine for children.

8. According to the preparation method of claim 7, characterized in that the pH regulator is a 0.5 - 56 wt% sodium hydroxide solution, preferably a 10 wt% sodium hydroxide solution. Preferably, the step of preparing the pH regulator includes: weighing the corresponding amount of sodium hydroxide, adding water for injection at 40 - 50 °C to make up the volume, stirring to dissolve it to make the concentration 10 wt%, cooling to below 35 °C and using it as the pH regulator for standby.

9. According to the preparation method of claim 7, characterized in that before preparing the liquid medicine, it also includes: checking the endotoxin of the water for injection to meet the standard of water for injection. Preferably, during the addition of sodium hydroxide, cool water is introduced into the jacket of the container to cool the solution in the container to below 25 °C. Preferably, before feeding, the jacket of the container is purged to drain the water in the jacket.

10. According to the preparation method of claim 7, characterized in that in the sterilization filtration step, pre-filtration and terminal sterilization filtration are adopted. For example, use a 0.45 μm pre-filter with a filter element made of polyethersulfone, a 0.22 μm redundant filter and a 0.22 μm terminal sterilization filter in series, and control the sterilization pressure ≤ 1.5 bar for sterilization operation. Preferably, the time interval from the start of liquid medicine preparation to the start of sterilization filtration shall not exceed 8 hours. Optionally, the time from the start of sterilization filtration to the end of sterilization filtration of the liquid medicine does not exceed 5 hours. Preferably, in the filling step, according to the single-dose filling amount required for clinical use, add water for injection below 30 °C to make up the volume. During the volume-making period, the magnetic pump stops stirring. After the volume-making is completed, start stirring, stir for 15 minutes, the frequency of the magnetic pump during stirring is 95%, and the temperature of the solution after volume-making is below 30 °C; take samples, detect the intermediate, and after sampling, close the magnetic pump and let the liquid medicine stand still.

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