CN110898040A - Freeze-dried powder for aztreonam solution for inhalation and preparation method thereof - Google Patents
Freeze-dried powder for aztreonam solution for inhalation and preparation method thereof Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/425—Thiazoles
- A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
- A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
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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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/007—Pulmonary tract; Aromatherapy
- A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
- A61K9/0078—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a nebulizer such as a jet nebulizer, ultrasonic nebulizer, e.g. in the form of aqueous drug solutions or dispersions
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
Abstract
The invention belongs to the field of pharmaceutics, and particularly relates to aztreonam solution freeze-dried powder for inhalation, and a preparation method and application thereof. The aztreonam solution for inhalation comprises freeze-dried powder for aztreonam inhalation and diluent, wherein the freeze-dried powder for inhalation and the diluent are mixed when in use. The aztreonam dry powder for inhalation provided by the invention has the advantages of good re-solubility, no need of other pH regulators and cosolvents, high stability, low anaphylaxis rate and the like.
Description
Technical Field
The invention belongs to the technical field of medicines, and relates to aztreonam solution freeze-dried powder for inhalation and a preparation method thereof.
Background
Aztreonam (Aztreonam) is an antibacterial drug developed by Shinpao corporation in America, is marketed in Italy in 1984, is the first monocyclic β -lactam antibiotic applied to clinic, has stronger antibacterial activity on gram-negative bacilli, and is stable to β -lactam enzyme.
In 2010, aztreonam-lysine inhalation was marketed as approved by the FDA and can be inhaled by patients by nebulization to exert therapeutic effects, and is made by a) dissolving α form of aztreonam in water to form a slurry, b) titrating the aqueous lysine solution to form aztreonam lysine salt in solution, c) drying the aztreonam lysine salt from the solution by freeze-drying or spray-drying, wherein the aztreonam lysine salt is milled, precipitated, spray-dried or otherwise processed to a particle size of 1-5 microns.
Chemical name of aztreonam: [2S- [2a, 3b (Z) ] ] -2- [ [ [1- (2-amino-4-thiazolyl) -2- [ (2-methyl-4-oxo-1-sulfo-3-azetidinyl) amino ] -2-oxyethylene ] amino ] oxo ] -2-methylpropionic acid. CAS number: 78110-38-0, molecular weight: 435.43, respectively; the structure is as follows:
aztreonam is a white or yellowish polycrystalline powder, the aztreonam raw material has four crystal forms, namely α crystal form, β crystal form, gamma crystal form and delta crystal form, the β crystal form is an aqueous crystal form, usually contains 7-14% of water, is easy to dissolve in water, is easy to absorb moisture, is poor in stability, is easy to open a ring under the conditions of moisture and heat, and forms open-ring aztreonam, the open-ring aztreonam is a main impurity of the aztreonam, the content of the aztreonam is reduced due to the existence of the aztreonam, so that the potency of a medicament is reduced, and the sterilization and bacteriostasis effects of the aztreonam are reduced.
Therefore, the aztreonam lysine salt inhalation solution in the prior art has the following problems that aztreonam is easy to agglomerate at the bottom and the wall of a preparation container when an β type aztreonam aqueous solution is prepared, the aztreonam is difficult to dissolve, the aztreonam aqueous solution is not uniformly mixed with auxiliary materials, insoluble particles and unqualified clarity are not obtained when the aztreonam freeze-dried powder is used for injection or is redissolved, and the aztreonam freeze-dried powder is unstable in the preparation, storage and transportation processes.
Disclosure of Invention
The invention firstly provides freeze-dried powder for an aztreonam solution for inhalation, and an atomized inhalation solution preparation comprises aztreonam lysine dry powder which is prepared from β crystal forms of aztreonam and L-lysine in a molar ratio of 1: 1.5-1: 2 (such as 1:1.55, 1:1.6, 1:1.7, 1:1.8, 1:1.85 and 1: 1.9).
Further, as a preferred embodiment, the aztreonam lysine dry powder is prepared from β crystal forms of aztreonam and L-lysine in a molar ratio of 1: 1.7-1: 2, and the aztreonam lysine freeze-dried powder is good in re-solubility, convenient to store and transport and capable of effectively reducing endogenous allergens of aztreonam.
Further, as a preferred embodiment, the dilution of the lyophilized powder for aztreonam solution for inhalation is 1.5-3mg/mL (such as 2.0mg/mL, 2.5mg/mL, 3.0mg/mL) of sodium chloride aqueous solution.
The invention also provides a preparation method of the freeze-dried powder for preparing the aztreonam solution for inhalation, which sequentially comprises the following steps:
the liquid preparation method comprises the steps of adding L-lysine and β crystal form aztreonam into water, and completely dissolving to obtain a mixed liquid, wherein the mixed liquid is filtered, filled and half-filled to obtain a pre-freeze-dried solution;
a freeze-drying step: the temperature is uniformly reduced to minus 40 +/-10 ℃ from the temperature of the prepared solution within 1 hour, and the temperature is kept for 3 to 6 hours; after primary drying, setting the vacuum of a freeze-drying box at 0.150-0.3mbar, uniformly raising the temperature of a shelf from minus 40 ℃ plus or minus 10 ℃ to minus 20 ℃ plus or minus 5 ℃, and preserving the heat for 8-10 hours; the temperature of the shelf is uniformly increased from minus 20 +/-5 ℃ to minus 5 ℃, and the temperature is kept for 8 hours; raising the temperature of the shelf to 8 +/-2 ℃ at a constant speed, and keeping the temperature for 3-4 hours, wherein the temperature of the sample is lower than 8 ℃.
Further preferred is a lyophilization step: setting the shelf for 1h, uniformly cooling the temperature of the mixed solution to minus 40 +/-10 ℃, and preserving the heat for 3-6 h; primary drying: the preparation time of the condenser is 20-40 minutes, and the vacuum of a freeze-drying box is set to be 0.150-0.3 mbar; when in use, the shelf temperature is increased from minus 40 +/-10 ℃ to minus 20 +/-5 ℃ at a constant speed within 2-4 hours, the vacuum of a freeze-drying box is 0.150-0.3mbar, and the heat is preserved for 8-10 hours; when the freeze-drying agent is used, the temperature of a shelf is uniformly increased from minus 20 +/-5 ℃ to minus 5 ℃ within 2 hours, the vacuum of a freeze-drying box is 0.150-0.3mbar, and the heat is preserved for 8 hours; the shelf temperature is uniformly raised to 8 +/-2 ℃ in 3-4h, the freeze-drying box is vacuumized for 0.150-0.3mbar, the temperature is kept for 3-4h, and the sample temperature is lower than 8 ℃.
Further, the preparation method also comprises the step of filling nitrogen for tamponade at the temperature of 8 ℃.
Further, as a preferred embodiment, in the step of preparing the liquid, the β crystal form aztreonam and the L-lysine are uniformly mixed, added into water, and completely dissolved to obtain a mixed liquid.
Further, as a preferred embodiment, in the liquid preparation step, ① prepares 1/4-2/5 prescription amount of β crystal form aztreonam and water for injection into an aztreonam suspension, prepares L-lysine and water for injection into a lysine solution, slowly injects the lysine solution into the aztreonam solution until β crystal form aztreonam is completely dissolved to obtain a composite solution, ② adds 1/4-2/5 prescription amount of β crystal form aztreonam into the composite solution to completely dissolve the aztreonam, and ③ repeats ② steps until the prescription amount of aztreonam is completely dissolved to obtain a mixed liquid.
Further, as a preferred embodiment, the liquid preparation step is carried out at 2 to 20 ℃ (e.g., 3 ℃, 4 ℃, 5 ℃, 6 ℃, 7 ℃, 8 ℃, 9 ℃, 10 ℃, 11 ℃, 12 ℃, 13 ℃, 14 ℃, 15 ℃, 16 ℃, 17 ℃, 18 ℃, 19 ℃).
Further, as a preferred embodiment, the liquid preparation step is carried out at 2 to 8 ℃.
Further, in a preferred embodiment, in the solution preparation step, the mixed solution is filtered through 0.45 μm and 0.22 μm filters in this order.
The invention overcomes the problem of poor solubility of β crystal form, and effectively reduces the content of high polymer or high molecular impurities formed by ring-opening aztreonam, thereby reducing the allergic rate, and more specific beneficial effects of the invention are reflected in the following 5 aspects:
firstly, the freeze-dried powder for the aztreonam solution for inhalation and the preparation method thereof do not involve adding other acid-base regulators, and only use L-lysine to regulate the solubility and pH value of the aztreonam in the production process.
Secondly, the freeze-dried powder for the aztreonam solution for inhalation provided by the invention does not need to be added with other solubilizers before use.
In a third aspect, the freeze-dried powder for the aztreonam solution for inhalation provided by the invention, wherein the β crystal forms aztreonam and/or L-lysine are both sterile and pyrogen-free, and has the advantages of simple preparation process and low production cost.
In a fourth aspect, the freeze-dried powder for the aztreonam solution for inhalation has low water content and stable properties; moreover, the mixture is uniform, and the difference of clarity, pH value and irritation between bottles caused by incomplete dissolution can be avoided.
In a fifth aspect, the β crystal forms of aztreonam and L-lysine in the freeze-dried powder for the aztreonam solution for inhalation have stable properties in the processes of storage and transportation, the re-solubility of the aztreonam and L-lysine is equivalent to α aztreonam lysine salt, but the stability is obviously improved, and the allergic rate is obviously reduced.
Detailed Description
The preparation process and the materials used in the preparation or the dosage of the materials used in the preparation in the following examples of the pharmaceutical preparation are not limited to the words, and all methods containing the pharmaceutical preparation provided by the present invention are within the protection scope of the present invention.
1. The aztreonam used in the invention is in an aztreonam β crystal form, and the lysine used in the invention is L-lysine.
2. Both comparative sample 1 and comparative sample 2 aztreonam are crystalline forms of aztreonam α.
3. The aztreonam β crystal form starting material was prepared from shandong luoxin pharmaceutical industries, inc, aztreonam α crystal form.
4. Clarity examination: clarity and color of the solution: adding water according to the marked amount to obtain a solution containing 0.1g of aztreonam in each 1ml, wherein the solution is clear and colorless; if the liquid becomes turbid, the liquid is not concentrated as compared with the No. 1 turbidity standard liquid (appendix IXB of the second part of the 2005 edition of Chinese medicated leaven); if the color is developed, the color should not be darker than that of yellow or yellow-green No. 4 standard colorimetric solution (appendix IXA first method of the second part of Chinese pharmacopoeia 2005 edition).
5. Inspection of insoluble microparticles: according to the method of the appendix IXH of the second part of the Chinese pharmacopoeia 2005 edition.
6. Visual foreign matter inspection: IXC method according to appendix IXC of second part of Chinese pharmacopoeia 2005 edition
7. Checking liquid chromatography conditions of ring-opening aztreonam impurities and related substances: octadecylsilane chemically bonded silica is used as a filling agent; 0.02mol/L potassium dihydrogen phosphate solution (pH is adjusted to 3.0 +/-O.1 by 10 percent phosphoric acid) and methanol (75/25) are taken as a mobile phase, the detection wavelength is 254nm, the column temperature is 40 ℃, and the number of theoretical plates is not lower than 2000 according to the aztreonam peak; the separation degree between the ring-opened aztreonam and adjacent impurities is in accordance with the requirement.
Examples 1,
Lyophilized powder for aztreonam lysine inhalation (1000 lyophilized powder injections) consists of:
| β crystalline form aztreonam | 75g(0.172mol) |
| L-lysine | 46.7g(0.319mol) |
| Water for injection to | 1000ml |
Examples 2,
Lyophilized powder for aztreonam lysine inhalation (1000 lyophilized powder injections) consists of:
| β crystalline form aztreonam | 75g(0.172mol) |
| L-lysine | 37.1g(0.258mol) |
| Water for injection to | 1000ml |
Examples 3,
Lyophilized powder for aztreonam lysine inhalation (1000 lyophilized powder injections) consists of:
| β crystalline form aztreonam | 75g(0.172mol) |
| L-lysine | 50.3g(0.344mol) |
| Water for injection to | 1000ml |
Adding L-lysine and β crystal form aztreonam into water, dissolving completely to obtain a mixed solution, filtering the mixed solution, filling, and adding a half plug to obtain a pre-freeze-dried solution, wherein the whole process is carried out at 20 ℃.
A freeze-drying step: the temperature is uniformly reduced to minus 40 +/-10 ℃ from the temperature of the prepared solution within 1 hour, and the temperature is kept for 3 to 6 hours; after primary drying, setting the vacuum of a freeze-drying box at 0.150-0.3mbar, uniformly raising the temperature of a shelf from minus 40 ℃ plus or minus 10 ℃ to minus 20 ℃ plus or minus 5 ℃, and preserving the heat for 8-10 hours; the temperature of the shelf is uniformly increased from minus 20 +/-5 ℃ to minus 5 ℃, and the temperature is kept for 8 hours; raising the temperature of the shelf to 8 +/-2 ℃ at a constant speed, and keeping the temperature for 3-4 hours, wherein the temperature of the sample is lower than 8 ℃.
The above three prescription samples with different ratios are re-dissolved with 1ml of 0.17% sodium chloride solution, and the impurities and related substances are checked, and the average results of the checks are shown in table 1:
TABLE 1 results of examination of impurities and related substances after reconstitution of lyophilized powders for inhalation obtained by different formulations
The lyophilized samples for inhalation of different formulations were reconstituted with 1ml of 0.17% sodium chloride solution, and the reconstituted samples were examined for clarity, visible impurities, and insoluble particles, the results of which are shown in Table 2.
TABLE 2 investigation of visible foreign bodies and insoluble particles after reconstitution of lyophilized samples Nos. 1-5 in sequence
| Examination item | Example 1 | Example 2 | Example 3 |
| Visible foreign body | Qualified | Qualified | Qualified |
| Insoluble particle > 10 mu m (one/mL) | 9.3 | 10.2 | 9.8 |
| Insoluble particle > 20 μm (one/mL) | 0.8 | 0.9 | 0.5 |
The results in tables 1 and 2 show that different formulations have no effect on the stability and reconstitution of lyophilized powders for inhalation.
The following examination of the production method employed recipe 1.
Preparation method 1
Step a was carried out at 20 ℃:
sequence 1: adding aztreonam powder into lysine solution
Adding the L-lysine into 80% water for injection according to the prescription amount, stirring until the L-lysine is dissolved, adding aztreonam powder into the solution, and finishing the addition within 5 min.
Sequence 2: lysine solution is added to aztreonam solution
Adding the lysine with the prescription amount into 40 percent of water for injection under stirring, and stirring until the lysine is completely dissolved; the prescription amount of aztreonam is added into 40 percent of water for injection under stirring, the mixture is stirred until the mixture is completely suspended, and the lysine solution is slowly added into the aztreonam solution.
Sequence 3: lysine powder is added into aztreonam solution
Adding the prescribed amount of aztreonam into 80% water for injection under stirring, stirring until the aztreonam is completely suspended, adding the prescribed amount of lysine powder, and stirring until the aztreonam is completely dissolved.
Sequence 4: the aztreonam and lysine powder are mixed firstly and then dissolved into water for injection.
And uniformly mixing aztreonam and lysine in the mixing bag, slowly adding 80% of water for injection under a stirring state, and fixing the volume to the prescription amount after dissolution.
The sequence 5 is that aztreonam solution prepared from ① 1/4-2/5 prescription amount of aztreonam and water for injection is prepared into solution of lysine and water for injection, the solution is slowly injected into the aztreonam solution under the stirring state until the aztreonam is completely dissolved, ② is added with 1/4-2/5 aztreonam powder into the mixed solution of the aztreonam lysine, the lysine solution is added while stirring, ③ is repeated ② steps until the aztreonam is completely dissolved, and the water for injection is added to the full amount.
Step B
Putting the semi-finished product subpackaged in the step A on a shelf of a freeze-drying box, and closing a door of the freeze-drying box; prefreezing (cooling stage and cooling maintaining stage): setting the shelf for 1h, uniformly cooling the temperature of the mixed solution to minus 40 +/-10 ℃, and then keeping the temperature of minus 40 +/-10 ℃ for 3-6 h; primary drying: setting the vacuum of the freeze drying box at 0.150-0.3mbar for 20-40 min, raising the shelf temperature from-40 +/-10 deg.c to-20 +/-2 deg.c at constant speed for 2-4 hr, maintaining at-20 +/-2 deg.c for 8-10 hr, raising the shelf temperature from-20 +/-5 deg.c to-5 deg.c at constant speed for 2 hr, and maintaining at-5 deg.c for 8 hr; secondary drying: the temperature of the shelf is uniformly raised to 8 +/-2 ℃ within 3-4h, and then maintained at 8 +/-2 ℃ for 3-4h, wherein the temperature of the sample is lower than 8 ℃;
the compounding step for comparative sample 1 was also carried out at 20 ℃ and was prepared as described in AU2017200384a1 by a) dissolving aztreonam in the form of α in water to form a slurry, b) titrating the aqueous lysine solution to form aztreonam lysine salt in the solution, and the lyophilization process using the lyophilization process of page 26 second paragraph of AU2017200384a1, i.e. the rack of the lyophilizer was pre-cooled to 0 ℃ before the start of the operation to allow rapid freezing of the bulk solution in the vial, the lyophilized vial containing the bulk solution was placed on the pre-cooled rack of the lyophilizer, then the interior of the lyophilizer was cooled to-38 ℃ at which the temperature of the bulk solution in the vial was frozen, the temperature of the lyophilizer was maintained at-38 ℃ and the vacuum was adjusted to 0.08mbar within 3 hours at a constant rate, then the temperature of the lyophilizer was increased to-25 ℃ within a few minutes and maintained for about 11 hours, then the vacuum was adjusted to 0.047mbar within 3 hours and the temperature was increased to +50 ℃ and maintained for 12 hours and the vial was removed after 12 hours and the crimp was removed.
Multiple batches of serial No. 1 to 5 samples were lyophilized by step B, the lyophilized samples were reconstituted with 1ml of 0.17% sodium chloride solution, and the impurities and related substances were checked, and the average results of the checks are shown in table 3:
TABLE 3 results of inspection of impurities and related substances after reconstitution of lyophilized powder obtained at 20 deg.C
As can be seen from the results in table 3, only the sequence of step a is different, with sample No. sequence 1 having the highest total impurity content; the highest ring-opening impurities of sequence 1 and comparative 1 also rose the highest. The difference of clinical medication safety is caused by the difference of the fine product quality, and the characteristics of good stability, no need of pH adjustment and the like of the products obtained in the sequence 4 and the sequence 5 are highlighted only by the preparation and the freeze-drying.
And (3) carrying out freeze-drying on a plurality of batch sequence No. 1 to No. 5 samples through the step B, redissolving the freeze-dried samples by using 1ml of 0.17% sodium chloride solution, and carrying out the clarity, visible foreign matters and insoluble particles detection on the redissolved samples, wherein the detection results are shown in a table 4.
TABLE 4 investigation of visible foreign bodies and insoluble particles after reconstitution of lyophilized samples Nos. 1-5 in sequence
From Table 4, it can be seen that the re-solubility of the inventive sample and the comparative sample is equivalent at the compounding temperature of 20 ℃, which indicates that the β -aztreonam lysine obtained by the present invention has the solubility equivalent to that of the comparative sample α aztreonam lysine.
Preparation method II
The method comprises the following steps of adopting a first preparation method under the same conditions, wherein the difference is that the step of preparing the solution is carried out at 2-8 ℃, the comparison sample is a comparison sample 2 of aztreonam α crystal form, respectively detecting impurities and related substances of the sample obtained by the second preparation method and the comparison sample 2, the detection conditions are the same as the first preparation method, the detection results are shown in a table 5, and the detection results of visible foreign matters and insoluble particles are shown in a table 6:
TABLE 52-8 ℃ results of examination of impurities and related substances in the solution obtained in step A
Comparing the experimental results of tables 3 and 5, the open loop impurities and total impurities were highest for the comparative sample and sequence 1 compared to the samples obtained from the other sequences, both in the transverse direction under the same conditions and in the longitudinal direction at different temperatures. Different sequences of liquid preparation steps play a crucial role in the stability of the product.
Table 6 examination of visible foreign matter and insoluble particles after reconstitution of lyophilized samples nos. 1-5 in sequence and comparative sample No. 2
From table 6, it can be seen that the resolubility of the inventive sample and the comparative sample obtained at 2-8 ℃ is equivalent, indicating that the β -aztreonam lysine obtained by the invention has solubility equivalent to α aztreonam lysine.
Through multiple small-test pilot-scale prescription process researches, the sequence 2 and the sequence 3 both need to prepare the aztreonam with the prescription amount into solutions, the phenomena of adherence and sticking are serious in the preparation process of the small-test solutions, the mixing time is long, complete dissolution is difficult to ensure after amplification, and the mixing is uniform, so that the method is not suitable for large-scale production; the open-loop impurities of the product obtained in the liquid preparation sequence 1 are equivalent to those of the comparative sample, and finally the liquid preparation is determined to be carried out by adopting the sequence 4 or the sequence 5, and the freeze-drying process comprises the steps of putting the semi-finished product subpackaged in the step A on a shelf of a freeze-drying box, and closing a door of the freeze-drying box; prefreezing (cooling stage and cooling maintaining stage): setting the shelf to use 1h, uniformly reducing the temperature from 20-25 ℃ to minus 40 +/-10 ℃, and then keeping the temperature at minus 40 +/-10 ℃ for 3-6 h; primary drying: setting the vacuum of the freeze drying box at 0.150-0.3mbar for 20-40 min, raising the shelf temperature from-40 +/-10 deg.c to-20 +/-2 deg.c at constant speed for 2-4 hr, maintaining at-20 +/-2 deg.c for 8-10 hr, raising the shelf temperature from-20 +/-5 deg.c to-5 deg.c at constant speed for 2 hr, and maintaining at-5 deg.c for 8 hr; secondary drying: the temperature of the shelf is uniformly raised to 8 +/-2 ℃ within 3-4h, and then maintained at 8 +/-2 ℃ for 3-4h, wherein the temperature of the sample is lower than 8 ℃. "
The quality standard item investigation of the aztreonam lysine inhalation preparation freeze-dried powder and the comparison sample is carried out in all aspects, the investigation result shows that the aztreonam lysine inhalation preparation freeze-dried powder and the comparison sample do not show more remarkable difference at different liquid preparation temperatures of 20 ℃ and 2-8 ℃, the following experiment results are products obtained at optional liquid preparation temperatures of 20 ℃ or 2-8 ℃, the investigation conditions of the comparison sample are the same as those of the aztreonam lysine inhalation preparation freeze-dried powder and the comparison sample, and the experiment results show that the total impurities, especially the ring-opening impurities, of the comparison sample are smaller at 2-8 ℃, so that the comparison sample selected in the following experiment is carried out at 2-8 ℃, and the comparison sample or α -aztreonam lysine salt is used for representing in the following experiment results.
Only the experimental results of significant differences between the present invention and the comparative samples are described herein, and detailed descriptions under other quality criteria items are omitted.
The stability of the inventive and comparative samples after 24 months at room temperature showed more significant differences, and the examination results are shown in Table 7
TABLE 7 stability test results of the inventive and comparative samples after 24 months of storage
As can be seen from the examination results of table 7, the present invention has shown a significant difference in stability compared to the comparative sample when left for 24 months at room temperature.
The sequence 5 of the second preparation method of the invention and the comparative sample 2 are selected to carry out a whole body active allergy experiment and a skin passive allergy experiment, and the specific implementation scheme is as follows:
medicine for experiment and experimental animal
1. The water for injection is prepared by self.
The 2.0.17% sodium chloride solution is prepared into proper concentration with normal saline before each use.
3. Bovine serum albumin, Genview product, lot number FA016-25G, available from Biotechnology, Inc. of Beijing ancient cooking.
4. Trichosanthin is provided by Tianjin, institute of medicine and science.
5. Evans blue, Flaka import split charging, Shanghai chemical reagent purchase supply station split charging factory.
Wistar rats weighing 180-220 g, half male and half female, and the qualification number (J) 2017-.
7. White guinea pig, weight 400 + -20 g, male, certificate number (J) 2017-.
8. In the active allergy experiment, the large and small dose groups of aztreonam are 625mg/kg and 312.5mg/kg respectively; 625mg/kg corresponds to the maximum daily dose of 8g for a 70kg person.
9. In passive allergy test, the size dose of aztreonam is 750mg/kg and 375mg/kg respectively, and 750mg/kg is equivalent to the maximum daily dose of 8g for 70kg people.
Systemic active hypersensitivity test
Before the start of the experiment, animals were kept for 1 week in advance to observe their activity, 96 healthy male white guinea pigs were randomly divided into 8 groups of 16 animals, the first group was a physiological saline group, the second group was a positive control group (bovine serum albumin 25mg/kg), the third group was α -aztreonam lysine salt 312.5mg/kg, the fourth group was α -aztreonam lysine salt 625mg/kg, the fifth group was β -aztreonam lysine 312.5mg/kg, the sixth group was β -aztreonam lysine salt 625 mg/kg., and the four-day intraperitoneal injection was given to each group of guinea pigs once every two times, the volume was 0.5 mL/mL and 3 times in total, to sensitize the guinea pigs, then each group was divided into two groups of 14d and 21d after the 1-day sensitization, physiological saline was given to the ear vein, bovine serum albumin 50mg/kg, α -aztreonam lysine salt 312.5mg/kg, the concentration of the cattle serum albumin was given to each day was 125 mg/kg, the highest concentration of the arginine salt was given to the ear vein, the group was found to be a nasal allergy reaction was found to be a systemic anaphylactic reaction (nasal shock), the animals, the patients were observed to have a systemic anaphylactic reaction, a systemic anaphylactic reaction was observed to be 1-allergic reaction, a nasal reaction to be observed to be 1-250 mg/kg, a nasal allergy to be observed to be 1-250 mg nasal allergy to be a nasal allergy to be equal to be 1 mg.
TABLE 8 symptoms of anaphylaxis
| 0 normal | 7 shortness of breath | 14 gait instability |
| 1 Bu Ning (a Chinese character of Bu' an) | 8 urination | 15 jump |
| 2 vertical hair | 9 discharging manure | 16 wheezing |
| 3 trembling | 10 tear flow | 17 spasm of liver |
| 4 scratching nose | 11 dyspnea | 18 rotate |
| 5 sneezing | 12 wheeze sound | 19 tidal breathing |
| 6 cough | Purpura 13 | 20 death |
TABLE 9 evaluation criteria for systemic sensitization
| 0 | - | Negative allergic reaction |
| 1 to 4 symptoms | + | Weak positive of anaphylaxis |
| 5 to 10 symptoms | ++ | Positive allergic reaction |
| 11 to 19 symptoms | +++ | Strong positive of allergic reaction |
| 20 | ++++ | Very strong positive of allergic reaction |
The results of the systemic active hypersensitivity test are shown in Table 10
TABLE 10 test results of systemic active hypersensitivity of the products of the invention and the comparative samples
As can be seen from Table 10, the 0.17% sodium chloride group, the third group of α -aztreonam lysine salt 412.5mg/kg, the fifth group of β -aztreonam lysine 312.5mg/kg and the sixth group of β -aztreonam lysine salt 625mg/kg all did not show grade 1-20 reaction symptoms, and were judged as being negative for anaphylaxis, while the group of α -aztreonam lysine salt 625mg/kg showed positive and weak positive reactions after being stimulated on days 14 and 21, respectively.
Passive skin allergy test
Before the experiment, the animals were kept for 1 week in advance to observe their activity performance, and 54 healthy rats, male and female halves, were randomly divided into six groups, 9 rats in each group, the first group was 0.17% physiological saline, the second group was a positive control group (trichosanthin 1mg/ml)5mg/kg, the third and fourth groups were α -aztreonam lysine salt (75mg/ml and 150mg/ml)375mg/kg and 750mg/kg, respectively, and the fifth and sixth groups were β -aztreonam lysine (75mg/ml and 150mg/ml)375mg/kg and 750mg/kg, respectively.
Preparation of IsE-containing antiserum from rats 1 rat per group was injected intramuscularly with the above-mentioned doses of physiological saline, trichosanthin, α -aztreonam lysine salt, β -aztreonam lysine salt, 1 ml/rat, 1 time every other day, 3 times in total, 10 days after the last sensitization, 3% chloral hydrate was used for anesthesia, abdominal aorta was bled, centrifuged at 2000r/min for 10min, and the serum was separated and stored at-20 ℃ for use, according to the requirements of the same kind of passive skin allergy test of the injected rats.
The same kind of passive skin anaphylaxis experiment of rats the rest 48 healthy rats were collected, 8 rats in each group were prepared with skin 4mm × 4mm on the two sides of the dorsal midline of each group of rats about 1.5cm from the spinal column. The above-mentioned IgE-containing antiserum was diluted with physiological saline at a ratio of 1: 4 and 1: 8 into 2 dilutions, and the diluted antiserum was injected into the prepared skin area on the back of the rat at the left side (1: 4 ratio) and the right side (1: 8 ratio), one spot per side, and 0.1ml per spot. After 48h of sensitization, each group was stimulated by injecting 1ml of the same amount of the stimulating antigen as the sensitizing dose plus an equal amount of 0.8% evans dye into the tail vein. After 30min, each group of rats was sacrificed and the diameter of blue reaction spots in the inner layer of the skin was measured, and the diameter of irregular spots was the sum of the major diameter and the minor diameter, and those with a diameter of more than 5mm were judged to be positive, and the results are shown in Table 11.
TABLE 11 Passive skin allergy test results
As can be seen from Table 11, the rats in the 0.17% sodium chloride group, the third group of α -aztreonam lysine salt of 375mg/kg, the fifth group of β -aztreonam lysine of 375mg/kg and the sixth group of 750mg/kg dose groups have no blue reaction spots on the back skin, the positive control group has blue spot reactions on the skin at the injection site in different degrees, and the diameters of the blue spots on both sides (1: 4 or 1: 8 dilution) are more than 5mm, so that the positive reaction is judged.
Claims (10)
1. A freeze-dried powder for an aztreonam solution for inhalation is characterized in that the freeze-dried powder is prepared from β crystal form aztreonam and L-lysine in a molar ratio of 1: 1.5-1: 2.
2. A freeze-dried powder for an aztreonam solution for inhalation according to claim 1, characterized in that the dry powder is prepared from β crystal form aztreonam and L-lysine in a molar ratio of 1: 1.7-1: 2.
3. A lyophilized powder for aztreonam solution for inhalation according to claim 1 or 2, characterized in that the diluent of the formulation of the nebulized inhalation solution is 1.5-3.0mg/mL of aqueous sodium chloride solution.
4. A process for the preparation of a lyophilized powder for an aztreonam solution for inhalation according to any one of claims 1 to 3, characterized in that it comprises the following steps in sequence:
the liquid preparation method comprises the steps of adding L-lysine and β crystal form aztreonam into water, and completely dissolving to obtain a mixed liquid, wherein the mixed liquid is filtered, filled and half-filled to obtain a pre-freeze-dried solution;
a freeze-drying step: a freeze-drying step: the temperature is uniformly reduced to minus 40 +/-10 ℃ from the temperature of the prepared solution within 1 hour, and the temperature is kept for 3 to 6 hours; after primary drying, setting the vacuum of a freeze-drying box at 0.150-0.3mbar, uniformly raising the temperature of a shelf from minus 40 ℃ plus or minus 10 ℃ to minus 20 ℃ plus or minus 5 ℃, and preserving the heat for 8-10 hours; the temperature of the shelf is uniformly increased from minus 20 +/-5 ℃ to minus 5 ℃, and the temperature is kept for 8 hours; raising the temperature of the shelf to 8 +/-2 ℃ at a constant speed, and keeping the temperature for 3-4 hours, wherein the temperature of the sample is lower than 8 ℃.
5. The method of claim 4, wherein the lyophilization step comprises: setting the shelf for 1h, uniformly cooling the temperature of the mixed solution to minus 40 +/-10 ℃, and preserving the heat for 3-6 h; primary drying: the preparation time of the condenser is 20-40 minutes, and the vacuum of a freeze-drying box is set to be 0.150-0.3 mbar; when in use, the shelf temperature is increased from minus 40 +/-10 ℃ to minus 20 +/-5 ℃ at a constant speed within 2-4 hours, the vacuum of a freeze-drying box is 0.150-0.3mbar, and the heat is preserved for 8-10 hours; when the freeze-drying agent is used, the temperature of a shelf is uniformly increased from minus 20 +/-5 ℃ to minus 5 ℃ within 2 hours, the vacuum of a freeze-drying box is 0.150-0.3mbar, and the heat is preserved for 8 hours; the shelf temperature is uniformly raised to 8 +/-2 ℃ for 3-4h, and the temperature is kept for 3-4h, wherein the sample temperature is lower than 8 ℃.
6. The preparation method according to claim 4 or 5, wherein the liquid preparation step comprises the steps of uniformly mixing β crystal form aztreonam and L-lysine, adding the mixture into water, and completely dissolving the mixture to obtain a mixed liquid.
7. The preparation method according to claim 4 or 5, characterized in that the liquid preparation step comprises the steps of ① preparing 1/4-2/5 prescription amount of β crystal form aztreonam and water for injection into an aztreonam suspension, preparing a lysine solution from prescription amount of L-lysine and water for injection, slowly injecting the lysine solution into the aztreonam solution until the β crystal form aztreonam is completely dissolved to obtain a mixed solution, ② adding 1/4-2/5 prescription amount of β crystal form aztreonam into the mixed solution to completely dissolve the aztreonam, and ③ repeating the ② step until the prescription amount of aztreonam is completely dissolved.
8. The method according to any one of claims 4 to 6, wherein the step of preparing the solution is carried out at 2 to 20 ℃.
9. The method according to any one of claims 4 to 6, wherein the step of preparing the solution is carried out at 2 to 8 ℃.
10. The method according to any one of claims 4 to 9, wherein the method further comprises a nitrogen-charging and pressure-plugging step at 8 ℃.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002051356A2 (en) * | 2000-12-27 | 2002-07-04 | Salus Pharma, Inc. | Inhalable aztreonam for treatment and prevention of pulmonary bacterial infections |
| WO2004052333A1 (en) * | 2002-12-11 | 2004-06-24 | Pari Gmbh | Pharmaceutical compositions for the pulmonary delivery of aztreonam |
| WO2005005424A1 (en) * | 2003-07-02 | 2005-01-20 | TEVA Gyógyszergyár Részvénytársaság | Aztreonam l-lysine and methods for the preparation thereof |
| CN101172974A (en) * | 2007-11-16 | 2008-05-07 | 西南合成制药股份有限公司 | Method for producing aztreonam amino acid salt |
| CN101579336A (en) * | 2009-07-07 | 2009-11-18 | 重庆市庆余堂制药有限公司 | Aztreonam for injection and production method thereof |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002051356A2 (en) * | 2000-12-27 | 2002-07-04 | Salus Pharma, Inc. | Inhalable aztreonam for treatment and prevention of pulmonary bacterial infections |
| WO2004052333A1 (en) * | 2002-12-11 | 2004-06-24 | Pari Gmbh | Pharmaceutical compositions for the pulmonary delivery of aztreonam |
| WO2005005424A1 (en) * | 2003-07-02 | 2005-01-20 | TEVA Gyógyszergyár Részvénytársaság | Aztreonam l-lysine and methods for the preparation thereof |
| CN101172974A (en) * | 2007-11-16 | 2008-05-07 | 西南合成制药股份有限公司 | Method for producing aztreonam amino acid salt |
| CN101579336A (en) * | 2009-07-07 | 2009-11-18 | 重庆市庆余堂制药有限公司 | Aztreonam for injection and production method thereof |
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