CN112361726B - Freeze-dried preparation water control method and application thereof - Google Patents
Freeze-dried preparation water control method and application thereof Download PDFInfo
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B5/00—Drying solid materials or objects by processes not involving the application of heat
- F26B5/04—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum
- F26B5/06—Drying solid materials or objects by processes not involving the application of heat by evaporation or sublimation of moisture under reduced pressure, e.g. in a vacuum the process involving freezing
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/66—Phosphorus compounds
- A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/55—Protease inhibitors
- A61K38/57—Protease inhibitors from animals; from humans
- A61K38/58—Protease inhibitors from animals; from humans from leeches, e.g. hirudin, eglin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F26—DRYING
- F26B—DRYING SOLID MATERIALS OR OBJECTS BY REMOVING LIQUID THEREFROM
- F26B25/00—Details of general application not covered by group F26B21/00 or F26B23/00
- F26B25/22—Controlling the drying process in dependence on liquid content of solid materials or objects
Abstract
The invention relates to a freeze-dried preparation water control method and application thereof, belonging to the technical field of preparation of freeze-dried preparations. The invention obtains the freeze-dried preparation with low impurity content and low water content by controlling the parameters of various operations of cooling speed, annealing temperature, annealing times, sublimation drying temperature and sublimation drying vacuum degree in the process of pre-freezing the preparation.
Description
Technical Field
The invention relates to the technical field of freeze-dried preparation, in particular to a freeze-dried preparation water control method and application thereof.
Background
The freeze-drying technology is that the material containing a large amount of water is cooled and frozen into solid in advance, and then solid ice crystals are directly sublimated out under the vacuum condition, so that the material per se keeps the original skeleton structure, the material keeps the original shape, and the rehydration of the product is excellent.
The freeze-drying technique has many different applications, and is indispensable in many biochemical and pharmaceutical applications, and the freeze-drying method is used for drying products such as pharmaceutical preparations and the like under vacuum, so that the products are always in a high-oxygen-deficient state, the easily oxidized medicines are protected and are not oxidized, and biological materials which can be stored for a long time and are not easy to deteriorate, such as microbial cultures, enzymes, blood and medicines, are obtained, and besides the stability of long-term storage, the freeze-drying method also retains the inherent biological activity and structure.
Generally, the properties of the raw material medicines of the freeze-dried preparation are unstable and are easy to degrade when exposed to damp and heat, and the raw material medicines or the medicine preparation are developed into the freeze-dried preparation due to the unstable characteristics of the raw material medicines when exposed to damp and heat. The lower the moisture content in the finished lyophilized preparation, the more stable the drug can be stored for a long time, so the control of the moisture of the product is particularly critical in the preparation process of the lyophilized preparation. Most of freeze-dried preparations on the market at present have large mass change during storage, are easy to degrade during storage, and have short effective period, which is caused by high water content of products. Therefore, in order to improve the shelf life of the lyophilized preparation, it is necessary to minimize the moisture content of the lyophilized preparation during the preparation process of the lyophilized preparation, thereby improving the stability and shelf life of the lyophilized preparation.
Disclosure of Invention
The invention aims to provide a freeze-dried preparation water control method and application thereof, which can effectively reduce the water content in a freeze-dried preparation, keep the drug effectiveness of the freeze-dried preparation and prolong the storage and quality guarantee period of the freeze-dried preparation.
In the research process of the long-term freeze-drying process, the strategy of slow cooling and multiple annealing is adopted in the pre-freezing stage of the preparation of the freeze-dried preparation, the annealing temperature is controlled to be 2-5 ℃ higher than the eutectic point, particularly 2 ℃ higher than the eutectic point, the annealing is carried out for 2-3 times, the temperature is kept for 50-90min (preferably 1h) each time, the crystal size is uniformly distributed, the later-stage sublimation drying is facilitated, the temperature of the preparation is controlled to be 2-5 ℃ lower than the eutectic point in the sublimation drying process, and the vacuum degree of a box body in the sublimation drying process is controlled to be 20 +/-3 Pa so as to achieve the higher sublimation drying speed and effectively reduce the moisture content in the freeze-dried preparation. The annealing time refers to the time of heat preservation after the annealing temperature is raised to-28 ℃ to-20 ℃.
The eutectic point is the temperature at which the water in the material is completely frozen into ice crystals. The invention discovers that annealing is carried out for 2-3 times at the temperature of 2-5 ℃ above the eutectic point temperature, the temperature is raised to-28-minus 20 ℃ in the annealing stage, the temperature is kept for 50-90min, particularly, the temperature is raised to-25-minus 21 ℃, the temperature is kept for 50-80 min, and the temperature of the product is kept to be 2-5 ℃ lower than the eutectic point temperature in sublimation, so that the moisture of the product can be removed, and the stability of the product can be improved. Through a large number of experiments, the technical scheme is determined, so that the prepared freeze-dried preparation obtains ideal crystals in the pre-freezing stage, the sublimation drying speed is improved, and the products are prevented from melting and collapsing.
Specifically, the invention provides a water control method of a freeze-dried preparation, which comprises the steps of slowly cooling in a pre-freezing stage of the preparation, preserving heat for a period of time when the temperature is reduced to-50 ℃ to-40 ℃, and then entering an annealing stage, wherein the annealing temperature in the annealing stage is 2-5 ℃ higher than the eutectic temperature of the preparation, and annealing is carried out for 2-3 times, and the temperature is kept for 50-90min each time. Preferably, after annealing to the target temperature, the temperature is maintained for 1 hour.
The temperature reduction speed of slow temperature reduction in the pre-freezing stage is 8-13 ℃/h, and preferably 9-11 ℃/h.
The method for controlling the moisture of the freeze-dried preparation is particularly suitable for freeze-drying preparation of the liquid medicine preparation which takes mannitol as a propping agent and has the solid content of the liquid medicine between 5.0 percent and 20.0 percent; the% is the mass-to-volume ratio of solid content to liquid (w/v, mg: ml) in the preparation.
In the water control method provided by the invention, the annealing stage comprises the steps of heating the preparation which is kept at the temperature of-50 to-40 ℃ for a period of time to the temperature of-28 to-20 ℃ at the speed of 15 to 25 ℃/h, and keeping the temperature for 50 to 90 min; and after the annealing is finished, cooling to-50 to-40 ℃ at the speed of 8-13 ℃/h, and preserving the heat for 2-2.5 h. Preferably, after the first annealing is finished, the temperature is reduced to minus 50 to minus 40 ℃, the heat preservation is carried out for 2 to 2.5 hours, and after the last annealing is finished, the temperature is reduced to minus 50 to minus 40 ℃, and the heat preservation is carried out for 3 to 5 hours.
In the pre-freezing stage of the method, after the temperature is slowly reduced to the target temperature, the heat preservation for a period of time means heat preservation for 1-4 hours.
The invention provides a method for controlling the moisture of the freeze-dried preparation, wherein the annealing stage comprises the steps of heating the preparation which is kept at the temperature of-50 to-40 ℃ for 1 to 3 hours to-25 to-21 ℃ at the speed of 18 to 22 ℃/h, and keeping the temperature for 50 to 80 min; cooling to-50 to-40 ℃ at the speed of 9-12 ℃/h, and preserving heat for 2-5 h; preferably, the temperature is increased to-25 to-22 ℃ at the speed of 20 ℃/h, and the temperature is kept for 60 min; and then cooling to-50 to-40 ℃ at the speed of 10 ℃/h, and preserving heat for 2 to 3 h.
The method further comprises a sublimation drying stage after annealing is finished, so that the vacuum degree reaches 17-25 Pa, the temperature of the partition plate is increased to 2-5 ℃ below the eutectic point temperature of the preparation at a speed of 4-6 ℃/h, and the temperature is kept for 6-12 h. Preferably, the incubation is carried out for 8 hours.
The method for controlling the moisture of the freeze-drying preparation further comprises the step of entering an analysis drying stage after sublimation drying, wherein a partition plate is arranged to raise the temperature to 38-45 ℃ at the speed of 18-22 ℃/h, the temperature is kept for 4-6 h, and an air mixing valve is closed for the last 1.5-2.5 h, so that the freeze-drying box keeps the ultimate vacuum degree.
Then, after the freeze drying is finished, filling nitrogen into the freeze drying box to 50MPa, and pressing and plugging; rolling an aluminum-plastic combined cover to obtain a freeze-dried preparation product.
Therefore, the invention also provides a freeze-dried product prepared by the freeze-dried preparation water control method.
The invention also provides application of the freeze-dried preparation water control method in reducing the water content of the freeze-dried preparation and improving the storage stability of the preparation.
The invention has the beneficial effects that:
(1) the method adopts a strategy of slowly cooling in a pre-freezing stage, controls the cooling speed of 10 ℃/h, and reduces the temperature of the product to a target temperature to obtain larger crystals;
(2) the invention adopts a control strategy of annealing in a pre-freezing stage; controlling the annealing temperature to be 2 ℃ higher than the eutectic point temperature, annealing for 2-3 times, keeping for 50-90min each time, enabling the crystal size to be uniformly distributed, obtaining ideal crystals, and further facilitating later sublimation drying;
(3) according to the invention, the temperature of the product in the sublimation drying process is controlled to be 2-5 ℃ lower than the temperature of the eutectic point, and the vacuum degree of the box body in the sublimation drying process is controlled to be 20 +/-3 Pa, so that the sublimation drying speed is further improved, and the reduction of the product quality caused by melting and collapsing of the product is avoided;
(4) the freeze-dried preparation product prepared by the method has low moisture content, the moisture content of the prepared tedizolid phosphate freeze-dried preparation for injection is 0.84%, the moisture content of the prepared azacitidine freeze-dried preparation for injection is 0.56%, the moisture content of the prepared bivalirudin freeze-dried preparation for injection is 1.34%, the impurity content of the freeze-dried preparation for injection is lower than that of similar products sold in the market, the purity is higher, the stability is better, and the effective period is longer.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
In the following examples, tedizolid phosphate for injection (specification: 200mg, solid content of drug solution 20%) is exemplified in example 1 and example 2, azacitidine for injection (specification: 100mg, solid content of drug solution 5%) is exemplified in example 3 and example 4, and bivalirudin for injection (specification: 250mg, solid content of drug solution 20%) is exemplified in example 5 and example 6. The moisture content in the embodiment of the present invention is measured by the first method (fisher's method) of the determination of moisture content of the fourth 0832, which is published in the chinese pharmacopoeia 2015.
In the following examples and comparative examples of the invention, each parallel experiment was repeated 5 times, and the mean values were taken, and the data differences between the batches were not significant.
Tedizolid phosphate for injection (solid content of liquid medicine is about 20%, sodium hydroxide is used for adjusting pH value, and dosage is small, so that dosage can be ignored)
Azacitidine prescription composition for injection (solid content of liquid medicine is 5%)
Bivalirudin prescription for injection (solid content of liquid medicine is 20%)
Example 1 Tedizolid phosphate for injection (200mg) lyophilization process:
prefreezing strategy using slow cooling
(1) A preparation stage: putting the half-stoppered tedizolid phosphate sample into a freeze dryer;
(2) pre-freezing: setting the cooling speed of a freeze dryer to 10 ℃/h, and preserving heat for 3h when the temperature of the product is reduced to-40 ℃;
(3) sublimation drying: starting a vacuum pump, when the vacuum degree reaches 20Pa, setting a partition plate to heat up to-20 ℃ at the speed of 5 ℃/h, and preserving heat for 12 h;
(4) and (3) resolving and drying: setting a clapboard to heat up to 40 ℃ at the speed of 20 ℃/h, preserving the heat for 5h after the temperature of the clapboard reaches 40 ℃, and closing a gas mixing valve in the last 2h to ensure that the freeze-drying box keeps the ultimate vacuum degree;
(5) and (3) plug pressing: after the freeze drying is finished, filling nitrogen into the freeze drying box to 50MPa, and pressing and plugging;
(6) and (3) rolling a cover: and rolling the aluminum-plastic combined cover.
Example 2 Tedizolid phosphate for injection (200mg) lyophilization process:
prefreezing strategy with 2 anneals
(1) A preparation stage: putting the half-stoppered tedizolid phosphate sample into a freeze dryer;
(2) pre-freezing: setting the cooling speed of a freeze dryer at 10 ℃/h, preserving heat for 1h when the temperature of a product is reduced to-40 ℃, then heating to-23 ℃ at the speed of 20 ℃/h, preserving heat for 1h, then cooling to-40 ℃ at the speed of 10 ℃/h, preserving heat for 2h, repeating annealing for 2 times, and preserving heat for 3h at-40 ℃;
(3) sublimation drying: starting a vacuum pump, when the vacuum degree reaches 20Pa, setting a partition plate to heat up to-20 ℃ at the speed of 5 ℃/h, and preserving heat for 8 h;
(4) and (3) resolving and drying: setting a clapboard to heat up to 40 ℃ at the speed of 20 ℃/h, preserving the heat for 5h after the temperature of the clapboard reaches 40 ℃, and closing a gas mixing valve in the last 2h to ensure that the freeze-drying box keeps the ultimate vacuum degree;
(5) and (3) plug pressing: after the freeze drying is finished, filling nitrogen into the freeze drying box to 50MPa, and pressing and plugging;
(6) and (3) rolling a cover: and rolling the aluminum-plastic combined cover.
The tedizolid phosphate for injection prepared in the technical scheme of the embodiment 2 adopts a prefreezing process of slowly cooling and annealing for 2 times, the temperature of a product is controlled to be lower than the eutectic point temperature by 3 ℃ in the sublimation drying process, and the vacuum degree of a drying oven is kept at 20Pa enough to achieve a faster sublimation speed, and meanwhile, the preparation product has lower moisture and good stability during storage.
The stability was investigated while detecting moisture in examples 1 and 2 of the present invention and the commercially available tedizolid phosphate. The moisture test results are shown in table 1.
Information on tedizolid phosphate: the manufacturer: merck & co, inc. trade name: sivextro, lot number: G00256.
TABLE 1 moisture comparison results
| / | Example 1 | Example 2 | Tedizolid phosphate on sale |
| Moisture (%) | 1.26 | 0.84 | 3.42 |
The moisture detection results show that: the moisture content of the products of examples 1 and 2 is obviously lower than that of the product sold in tedizolid phosphate, the lower moisture content is beneficial to improving the stability of the products during storage, and the stability results are shown in a table 2.
TABLE 2 stability test results
The stability test result shows that: the tedizolid phosphate product for injection prepared by adopting the slow cooling and annealing process prefreezing of the embodiment 2 has low water content, good stability during storage, no obvious change in product quality, low water content, few impurities and high purity.
Example 3 azacitidine for injection (100mg) lyophilization process:
prefreezing strategy using slow cooling
(1) A preparation stage: placing the azacitidine sample after being filled with the half stopper into a freeze dryer;
(2) pre-freezing: setting the cooling speed of a freeze dryer to 10 ℃/h, and preserving heat for 3h when the temperature of the product is reduced to-45 ℃;
(3) sublimation drying: starting a vacuum pump, when the vacuum degree reaches 20Pa, setting a partition plate to heat up to-17 ℃ at the speed of 5 ℃/h, and preserving heat for 10 h;
(4) and (3) resolving and drying: setting a clapboard to heat up to 35 ℃ at the speed of 20 ℃/h, preserving the heat for 5h after the temperature of the clapboard reaches 35 ℃, and closing a gas mixing valve in the last 2h to ensure that the freeze-drying box keeps the ultimate vacuum degree;
(5) and (3) plug pressing: after the freeze drying is finished, filling nitrogen into the freeze drying box to normal pressure, and pressing and plugging;
(6) and (3) rolling a cover: and rolling the aluminum-plastic combined cover.
Example 4 azacitidine for injection (100mg) lyophilization process:
prefreezing strategy with 2 anneals
(1) A preparation stage: placing the azacitidine sample after being filled with the half stopper into a freeze dryer;
(2) pre-freezing: setting the cooling speed of a freeze dryer to 10 ℃/h, preserving heat for 1h when the temperature of a product is reduced to-45 ℃, then increasing the temperature to-23 ℃ at the speed of 20 ℃/h, preserving heat for 1h, then reducing the temperature value to-40 ℃ at the speed of 10 ℃/h, preserving heat for 2h, repeatedly annealing for 2 times in the way, and preserving heat for 3h at-40 ℃;
(3) sublimation drying: starting a vacuum pump, when the vacuum degree reaches 20Pa, setting a partition plate to heat up to-17 ℃ at the speed of 5 ℃/h, and preserving heat for 7 h;
(4) and (3) resolving and drying: setting a clapboard to heat up to 35 ℃ at the speed of 20 ℃/h, preserving the heat for 5h after the temperature of the clapboard reaches 35 ℃, and closing a gas mixing valve in the last 2h to ensure that the freeze-drying box keeps the ultimate vacuum degree;
(5) and (3) plug pressing: after the freeze drying is finished, filling nitrogen into the freeze drying box to normal pressure, and pressing and plugging;
(6) and (3) rolling a cover: and rolling the aluminum-plastic combined cover.
The azacitidine for injection prepared in the technical scheme of the invention adopts a prefreezing process of slowly cooling and annealing for 2 times, the temperature of the product is controlled to be lower than the eutectic point temperature by 5 ℃ in the sublimation drying process, and the vacuum degree of a drying oven is kept at 20Pa enough to achieve a faster sublimation speed, and meanwhile, the preparation product has lower moisture and good stability during storage.
The stability of azacitidine for injection in examples 3 and 4 of the present invention and on the market was investigated together with the water content. The results are shown in Table 3. Information on azacitidine sale: the manufacturer: trade name of Celgene Europe ltd: vidaza (vidasa), batch number: 8J 132A.
TABLE 3 moisture comparison results
| / | Example 3 | Example 4 | Azacitidine on sale |
| Moisture (%) | 0.63 | 0.56 | 2.54 |
The moisture detection results show that: the products of examples 3 and 4 had significantly lower moisture than azacitidine on the market and lower moisture was beneficial to improve the stability of the product during storage, and the results are shown in table 4.
TABLE 4 stability test results
The stability test result shows that: the azacitidine product for injection prepared by adopting slow cooling and annealing process prefreezing has low water content, good stability during storage, no obvious change in product quality, low water content, less impurities and high purity.
Example 5 bivalirudin for injection (250mg) lyophilization process:
prefreezing strategy using slow cooling
(1) A preparation stage: putting the bivalirudin sample after filling and half plugging into a freeze dryer;
(2) pre-freezing: setting the cooling speed of a freeze dryer to 10 ℃/h, and preserving heat for 3h when the temperature of the product is reduced to-50 ℃;
(3) sublimation drying: starting a vacuum pump, when the vacuum degree reaches 20Pa, setting a partition plate to heat to-25 ℃ at the speed of 5 ℃/h, and preserving heat for 15 h;
(4) and (3) resolving and drying: setting a clapboard to heat up to 35 ℃ at the speed of 20 ℃/h, preserving the heat for 8h after the temperature of the clapboard reaches 35 ℃, and closing a gas mixing valve in the last 2h to ensure that the freeze-drying box keeps the ultimate vacuum degree;
(5) and (3) plug pressing: after the freeze drying is finished, filling nitrogen into the freeze drying box to normal pressure, and pressing and plugging;
(6) and (3) rolling a cover: and rolling the aluminum-plastic combined cover.
Example 6 freeze-drying Process of bivalirudin for injection (250 mg):
prefreezing strategy with 2 anneals
(1) A preparation stage: putting the bivalirudin sample after filling and half plugging into a freeze dryer;
(2) pre-freezing: setting the cooling speed of a freeze dryer at 10 ℃/h, preserving heat for 1h when the temperature of a product is reduced to-50 ℃, then increasing the temperature to-25 ℃ at the speed of 20 ℃/h, preserving heat for 1h, then reducing the temperature to-50 ℃ at the speed of 10 ℃/h, preserving heat for 2h, repeatedly annealing for 2 times in the way, and preserving heat for 3h at-50 ℃;
(3) sublimation drying: starting a vacuum pump, when the vacuum degree reaches 20Pa, setting a partition plate to heat up to-25 ℃ at the speed of 5 ℃/h, and preserving heat for 7 h;
(4) and (3) resolving and drying: setting a clapboard to heat up to 35 ℃ at the speed of 20 ℃/h, preserving the heat for 8h after the temperature of the clapboard reaches 35 ℃, and closing a gas mixing valve in the last 2h to ensure that the freeze-drying box keeps the ultimate vacuum degree;
(5) and (3) plug pressing: after the freeze drying is finished, filling nitrogen into the freeze drying box to normal pressure, and pressing and plugging;
(6) and (3) rolling a cover: and rolling the aluminum-plastic combined cover.
The bivalirudin for injection prepared in the technical scheme of the invention adopts a prefreezing process of slowly cooling and annealing for 2 times, the temperature of the product is controlled to be 3 ℃ lower than the eutectic point in the sublimation drying process, and the vacuum degree of a drying oven can be kept at 20Pa enough to achieve a faster sublimation speed, and meanwhile, the preparation product has lower moisture and good stability during storage.
The results of the electron microscope scanning of example 5 and example 6 of the present invention and the commercial bivalirudin show that the crystal size is uniform and is an ideal crystal, and the stability study was performed while detecting moisture, and are shown in table 5.
In the context of the marketed bivalirudin information: the manufacturer: THE MEDICINES COMPANY, trade name: angiomax, lot number: 00142.
TABLE 5 moisture comparison results
| / | Example 5 | Example 6 | On sale bivalirudin |
| Moisture (%) | 1.65 | 1.34 | 2.96 |
The moisture detection results show that: the moisture content of the products of examples 5 and 6 was significantly lower than that of the products in the case of salebivalirudin, the lower moisture content was beneficial to improving the stability of the products during storage, and the stability results are shown in Table 6.
TABLE 6 stability test results
The stability test result shows that: the injection bivalirudin product prepared by adopting slow cooling and annealing process prefreezing has lower water content, good stability during storage, no obvious change in product quality, and the prepared freeze-dried product has the water content obviously lower than that of a commercial similar freeze-dried preparation, less impurity content and high purity.
Comparative example 1 formulation with glucose as proppant
The tedizolid phosphate preparation with glucose as the propping agent comprises 200mg of tedizolid phosphate, 200mg of glucose, a proper amount of sodium hydroxide and 2ml of water for injection.
Referring to the method of example 2 of the present invention, glucose was used as a proppant, and the freeze-drying process as in example 2 was used for the tedizolid phosphate preparation of this comparative example, and the obtained product had a large number of bubbles and bulges, and the product appearance was poor, and the reason for analysis was that the viscosity of glucose was high, and the moisture was not easily escaped during the drying process, and the final product had a high moisture content. The moisture test results are shown in Table 7.
TABLE 7 moisture comparison results
| / | Example 1 | Example 2 | Comparative example 1 | Tedizolid phosphate on sale |
| Moisture (%) | 1.26 | 0.84 | 4.87 | 3.42 |
Comparative example 2 the preparation has a liquid solid content of 2%, 22%, 25%, 10%
The preparation 1 of the comparative example is a azacitidine preparation for injection, and the prescription composition thereof comprises 100mg of azacitidine, 100mg of mannitol and 10ml of water for injection. Namely, the solid content of the liquid medicine in the preparation is 2 percent.
The preparation 2 of the comparative example is azacitidine preparation for injection, and the prescription composition of the preparation comprises 220mg of azacitidine, 220mg of mannitol and 2ml of water for injection. Namely, the solid content of the liquid medicine in the preparation is 22 percent.
The preparation 3 of the comparative example is azacitidine preparation for injection, and the prescription composition of the preparation comprises 280mg of azacitidine, 220mg of mannitol and 2ml of water for injection. Namely, the solid content of the liquid medicine in the preparation is 25 percent.
The preparation 4 of the comparative example is azacitidine preparation for injection, and the prescription composition of the preparation is 100mg of azacitidine, 100mg of mannitol and 2ml of water for injection. Namely, the solid content of the liquid medicine in the preparation is 10 percent.
The azacitidine liquid preparation with the solid content of 2%, 22%, 25% and 10% is tested, and after the freeze-drying process the same as that in the embodiment 4 is adopted, the obtained product of the preparation 1 is found to be loose and dispersed, part of samples fly out of the bottle in the freeze-drying process, the product appearance is poor, and the product is polluted and the content is unqualified due to the fact that part of the products fly out of the bottle; but the final product has a lower moisture content due to the lower solids content.
Formulation 2 and formulation 3 appeared as: the product is compact, the inner pore canal is small, the moisture is not easy to escape, the moisture of the product is high, and the bottom of the preparation 3 is slightly melted.
Formulation 4 appeared as: the product is in loose block shape, the inside is loose and porous, the moisture can escape favorably, and the moisture of the final product is lower. The azacitidine content and moisture measurements are shown in Table 8.
TABLE 8 moisture comparison results
Comparative example 3 annealing stage only 1 anneal
Bivalirudin prescription composition for injection
(1) A preparation stage: putting the bivalirudin sample after filling and half plugging into a freeze dryer;
(2) pre-freezing: setting the cooling speed of a freeze dryer at 10 ℃/h, preserving heat for 1h when the temperature of a product is reduced to-50 ℃, then heating to-25 ℃ at the speed of 20 ℃/h, preserving heat for 1h, then reducing the temperature value to-50 ℃ at the speed of 10 ℃/h, and preserving heat for 3h at-50 ℃;
(3) step (6) was the same as in example 6, and this comparative example was different from example 6 in that step (2) was annealed only once, and the resulting product had slight traces of melting at the bottom from a small amount, indicating failure of lyophilization.
Comparative example 4 extension of annealing incubation time
Bivalirudin prescription composition for injection
(1) A preparation stage: putting the bivalirudin sample after filling and half plugging into a freeze dryer;
(2) pre-freezing 1: setting the cooling speed of a freeze dryer at 10 ℃/h, preserving heat for 1h when the temperature of a product is reduced to-50 ℃, then heating to-25 ℃ at the speed of 20 ℃/h, preserving heat for 2h, then reducing the temperature value to-50 ℃ at the speed of 10 ℃/h, and preserving heat for 2 h; annealing for 2 times, and keeping the temperature at-50 ℃ for 3 h;
pre-freezing 2: the same method as pre-freezing 1 is adopted, except that the annealing time is kept for 40 min.
(3) Steps (6) are the same as in example 6.
This comparative example prefreeze 1 differs from example 6 in that: the holding time at-25 ℃ is prolonged from 1h to 2h during each annealing. The obtained product has fine and smooth appearance, a layer of hard shell is arranged on the surface of the product, and the moisture is not easy to escape, and the analysis reason is that the product is excessively melted in the annealing process due to the overlong annealing time, so that the product is in two cycles of freezing-complete melting-freezing-complete dissolving-freezing in the pre-freezing process, the aim of annealing to enable the ice crystal to grow cannot be achieved, the internal structure of the final product is fine and compact, the moisture is not easy to escape, and the moisture of the final product is high.
The product performance of prefreeze 2 is: this comparative example prefreeze 2 differs from example 6 in that: the heat preservation time at the temperature of 25 ℃ below zero is shortened from 1h to 40min during each annealing. The obtained product has fine and smooth appearance and is not beneficial to moisture escape, the analysis reason is that the annealing process is incomplete due to too short annealing time, the purpose of fully growing ice crystals by annealing cannot be achieved, the internal structure of the final product is fine and compact, the moisture escape is not facilitated, and the moisture of the final product is high. The results of the moisture detection are shown in Table 9.
TABLE 9 moisture comparison results
| / | Example 5 | Example 6 | Comparative example 4 prefreezing 1 | Comparative example 4 prefreezing 2 | On-sale bivalirudin |
| Moisture (%) | 1.65 | 1.34 | 3.18 | 2.87 | 2.96 |
Comparative example 5
The preparation of the comparative example is azacitidine preparation for injection, and the prescription composition of the preparation comprises 100mg of azacitidine, 100mg of mannitol and 4ml of water for injection.
According to the method of the embodiment 4 of the invention, the annealing temperature is changed to be higher than the eutectic point temperature by 6 ℃ only in the annealing stage, other process parameters are consistent with those of the embodiment 4, the obtained product has fine appearance, a layer of hard shell is arranged on the surface of the product, and moisture is not easy to escape, and the analysis reason is that the product is melted too fast in the annealing process and ice crystals cannot grow in time due to the fact that the annealing temperature is too high, so that the internal result of the product is fine and compact, moisture is not easy to escape, and the moisture of the final product is high. The results of the moisture measurements are shown in Table 10.
TABLE 10 moisture comparison results
| / | Example 3 | Example 4 | Comparative example 5 | Azacitidine on sale |
| Moisture (%) | 0.63 | 0.56 | 1.63 | 2.54 |
Comparative example 6
The preparation of the comparative example is azacitidine preparation for injection, and the prescription composition of the preparation comprises 100mg of azacitidine, 100mg of mannitol and 4ml of water for injection.
Referring to the method of the embodiment 4 of the invention, the sublimation temperature is changed to be lower than the eutectic point temperature by 1 ℃ only in the sublimation drying stage, other process parameters are kept consistent with those of the embodiment 4, the obtained product is cracked and broken in appearance, and the bottom of the product has obvious melting and collapse traces.
Although the invention has been described in detail with respect to the general description and the specific embodiments thereof, it will be apparent to those skilled in the art that modifications and improvements can be made based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (11)
1. A freeze-drying preparation water control method adopting slow cooling and multiple annealing is characterized by comprising a pre-freezing stage, an annealing stage, a sublimation drying stage after annealing is finished and an analysis drying stage after sublimation drying; the method sequentially comprises the following steps:
(1) a pre-freezing stage: in the pre-freezing stage of the preparation, a slow cooling strategy is adopted to obtain ideal larger crystals, the cooling speed of slow cooling in the pre-freezing stage is 9-11 ℃/h, the temperature is kept for a period of time when the temperature is reduced to minus 50 ℃ to minus 40 ℃, and then the annealing stage is carried out;
and (3) annealing stage: a multi-time annealing strategy is adopted in an annealing stage, the annealing temperature in the annealing stage is 2-5 ℃ higher than the eutectic point temperature of the preparation, annealing is carried out for 2-3 times, and the annealing is kept for 50-90min after each annealing to the target temperature; the eutectic point temperature is the temperature when water in the material is completely frozen into ice crystals; thereby ensuring the size distribution of the crystal to be uniform and being beneficial to the sublimation drying in the later period;
a sublimation drying stage: in the sublimation drying stage, the temperature of the preparation is controlled to be 2-5 ℃ lower than the eutectic point temperature of the preparation, and the vacuum degree of a box body in the sublimation drying stage process is controlled to be 20 +/-3 Pa so as to achieve a faster sublimation drying speed and effectively reduce the water content in the freeze-dried preparation; the sublimation drying speed is further improved, and the reduction of the product quality caused by melting and collapsing of the product is avoided;
and (3) analysis and drying stage: setting a partition plate, heating to 38-45 ℃ at a speed of 18-22 ℃/h, preserving heat for 4-6 h, and closing a gas mixing valve for the last 1.5-2.5 h to keep the freeze-drying box in a limiting vacuum degree;
the freeze-drying preparation water control method with slow cooling and multiple annealing can obtain ideal crystals in the freeze-drying process, so that the product is prevented from melting and collapsing; and improve the sublimation drying speed; the water content of the freeze-dried preparation can be effectively reduced;
the lyophilized preparation obtained by the method has low water content, low impurity content, high purity, and improved storage stability.
2. The method of claim 1, wherein the lyophilized formulation is tedizolid phosphate, azacitidine, or bivalirudin.
3. The moisture control method according to claim 1, wherein the preparation is a liquid medicine preparation which takes mannitol as a propping agent and has a solid content of 5.0-20.0%; the% is the mass to volume ratio of solid content to liquid (w/v).
4. The water control method according to any one of claims 1 to 3, wherein the annealing stage comprises heating the preparation, which is kept at-50 to-40 ℃ for a certain period of time, to-28 to-20 ℃ at a speed of 15 to 25 ℃/h, and keeping the temperature for 50 to 90 min; and after the annealing is finished, cooling to-50 to-40 ℃ at the speed of 8-13 ℃/h, and keeping the temperature for 2-5 h.
5. The method for controlling moisture according to claim 4, wherein the holding time is 1 to 4 hours.
6. The water control method according to any one of claims 1 to 3, wherein the annealing stage comprises heating the preparation, which is kept at-50 ℃ to-40 ℃ for 1-3 hours, to-25 ℃ to-21 ℃ at a speed of 18-22 ℃/hour, and keeping the temperature for 50-80 min; after the annealing is finished, cooling to-50 to-40 ℃ at the speed of 9-12 ℃/h, and preserving heat for 2-3 h.
7. The water control method according to claim 6, wherein the annealing stage comprises raising the temperature to-25 ℃ to-22 ℃ at a speed of 20 ℃/h, and keeping the temperature for 60 min; and then cooling to-50 to-40 ℃ at the speed of 10 ℃/h, and preserving heat for 2 to 3 h.
8. The moisture control method according to any one of claims 1 to 3, wherein after the annealing is finished, a sublimation drying stage is performed, the vacuum degree is increased to 17-25 Pa, the temperature is increased to 2-5 ℃ below the eutectic point temperature of the preparation at a speed of 4-6 ℃/h, and the temperature is maintained for 6-12 h.
9. The moisture control method according to any one of claims 1 to 3, further comprising, after the step (1) to the step (4), a step (5): after the freeze drying is finished, filling nitrogen into the freeze drying box to 50MPa, and pressing and plugging; rolling the aluminum-plastic combined cover to obtain the freeze-dried preparation product.
10. A lyophilized product produced by the moisture control method according to any one of claims 1 to 9.
11. Use of the moisture management method of any one of claims 1 to 9 for reducing the moisture content of a lyophilized formulation and improving the storage stability of the formulation.
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| CN113750054B (en) * | 2021-05-27 | 2023-03-24 | 北京康仁堂药业有限公司 | Preparation method of freeze-dried powder |
| CN113521019B (en) * | 2021-07-27 | 2023-08-18 | 吉林大学第一医院 | Mesenchymal stem cell supernatant freeze-dried preparation and preparation method thereof |
| CN115463128B (en) * | 2022-09-23 | 2023-10-03 | 深圳太太药业有限公司 | Panipenem and betamilone compound combination drug for injection and preparation method thereof |
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| CN107753516A (en) * | 2016-08-19 | 2018-03-06 | 泰凌(中国)投资有限公司 | A kind of Uropoly acid-peptide of low moisture absorption freezes compound powder and preparation method thereof |
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