CN110141561A - A kind of technology of preparing of aerosolizable protein dry powder inhalant - Google Patents
A kind of technology of preparing of aerosolizable protein dry powder inhalant Download PDFInfo
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- CN110141561A CN110141561A CN201810271022.XA CN201810271022A CN110141561A CN 110141561 A CN110141561 A CN 110141561A CN 201810271022 A CN201810271022 A CN 201810271022A CN 110141561 A CN110141561 A CN 110141561A
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- A—HUMAN NECESSITIES
- 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
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- A—HUMAN NECESSITIES
- 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
- 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
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
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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
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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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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
Abstract
The present invention discloses a kind of technology of preparing of aerosolizable protein dry powder inhalant.The present invention provides a kind of reagent for being used to prepare dry powder inhaler formulations, including solute, the solute is made of drug, mannitol, freeze drying protectant and dispersing agent；The freeze drying protectant is at least one of inositol, trehalose and dextran-40；The dispersing agent is made of leucine and poloxamer；The reagent is used into atomizing freeze drying (SFD) technology again, prepares the suitable dry powder inhaler formulations of partial size.The dry powder formulations are when storage and transport it is noted that moisture-proof with solidapowder form storage and transport, will affect dry powder aerosol when excessively moist and effect occurs.The dry powder formulations are soluble easily in water, and hygroscopicity is not strong, and water content is lower, and protein vigor loss is small in preparation process, and the dry powder aerosol occurred out can preferably be deposited on lower respiratory tract and lung, can play ideal effect for respiratory immune or respiratory administration.
Description
Technical field
The invention belongs to field of biotechnology more particularly to a kind of preparation skills of aerosolizable protein dry powder inhalant
Art.
Background technique
Most of strong pathogen can be sucked through aerosol to infect, and main infection position is respiratory tract and lung, compared to other
Route of infection spread speed is faster.Vaccine passes through traditional percutaneous or be inoculated with through intramuscular injection, respiratory tract and lung's mucosa-immune
Response is opposite to be lacked, so it is poor to the immune protective effect of aerosol sucking infection.The aerosolized transpulmonary suction of vaccine dry powder
Adaptive immune response is not only induced after entering immunity inoculation, also can induce respiratory tract and the lung of traditional immunization route of inoculation shortcoming
Mucosal immune response, so sucking protection against infection effect can be significantly improved.In addition, clinically certain infectious diseases and asthma etc.
Administration also often treated by way of respiratory tract administration, this administration mode can make drug go directly lesion, thus
Preferably play a role.
Compared to other routes of infection of pathogen, aerosol sucks transmission of infection speed faster, and disease is rapider, disease
Feelings are more serious, therefore most of strong pathogen for sucking infection body through aerosol are often considered as biological warfare agent and bio-terrorism
Agent.Vaccine inoculation is to defend the important means of biological warfare agent and bio-terrorism agent, but traditional percutaneous or be inoculated with through intramuscular injection,
Compared with direct lung's immunity inoculation, lack respiratory tract and lung's mucosal immune response, to the immune guarantor of aerosol sucking infection
Shield effect is poor, therefore lung's immunity inoculation is considered as the important immunization route for aerosol infection.Some researches show that powder
Agent can cause stronger mucosa-immune reaction compared with liquor, and reason may be that liquid vaccine is delivered after lung quickly by mucosal system
Removing campaign excrete, and the adhesive force that dry powder vaccine is delivered after lung in part is preferable, to delay mucous membrane pair
Its removing speed.Not only inducing systemic immune response after the transpulmonary inhalational immunization inoculation of aerosolized vaccine dry powder, usually may be used also
The respiratory tract and lung's mucosal immune response of traditional immunization route of inoculation shortcoming are induced, thus compared to traditional immunization inoculation side
Method, aerosolized vaccine dry powder can play more preferable or equal effect to sucking protection against infection effect.The suction of aerosolizable
Entering type dry powder vaccine preparation is a kind of vaccine preparation for being used directly for respiratory immunity, and ideal vaccine dry powder usually has
Water content is low, hygroscopicity is poor, good dispersion, property are stable, is easy to the advantages that transporting.
It prepares vaccine dry powder formulations and is related to the drying of liquid sample.Currently used dry technology has freeze-drying and spray
Mist drying etc..It is freeze-dried obtained pie product and usually obtains powder product by mechanical lapping, product cut size is distributed model
The heat generated when enclosing extensively, and grinding will cause product quality reduction.Spray drying will the sample that is directly atomized with hot gas
Solvent is evaporated during media contact, to obtain dried powder, but the active constituent in medical product is often due to thermo-labile
It is destroyed.The advantages of atomizing freeze drying method combines above two drying means, while them are overcome the shortcomings of again.It is spraying
Freezing dry process generally comprises following 3 steps: 1. using the atomizer of special designing the fluid sample mist for needing drying
Chemical conversion is tiny droplet；2. by cryogenic gas or liquid above-mentioned droplet being quickly cooled down and being freezed, is formed and freezed
Powder；3. carrying out vacuum freeze drying by sublimation principle to above-mentioned frozen powder, finally obtaining powdered drying finished product.
Research shows that 7 μm of particulate matters below of aerodynamic diameter can enter throat, 5 μm of particulate matters below can enter
The deep of respiratory tract, 2.5 μm of particles below as deep as alveolar and can deposit.
Summary of the invention
It is an object of the present invention to provide a kind of reagents for being used to prepare dry powder inhaler formulations.
Reagent provided by the invention, including solute,
The solute is made of drug, mannitol, freeze drying protectant and dispersing agent；
The freeze drying protectant is at least one of inositol, trehalose and dextran-40；
The dispersing agent is made of leucine and poloxamer.
In mentioned reagent,
The drug, the mannitol, the freeze drying protectant, the leucine and the poloxamer proportion be
0.05-0.15g:1g:1-1.5g:0.5-1g:0.05ml；
Or, the freeze drying protectant is inositol and trehalose, the mass ratio of the inositol and the trehalose is 1:1；
Or, the freeze drying protectant is inositol and dextran-40, the quality of the inositol and the dextran-40
Than for 1:0.5.
In mentioned reagent,
The reagent further includes solvent, and the solvent of solution can be deionized water, ultrapure water, distilled water etc..
The reagent is made of the solute and the solvent；
Quality volumn concentration of the drug in the reagent is 0.05-0.15%；
Quality volumn concentration of the mannitol in the reagent is 1%；
The freeze-dried quality volumn concentration in the reagent is 1-1.5%；
Quality volumn concentration of the inositol in the reagent is 0.5-1%；
Quality volumn concentration of the trehalose in the reagent is 0.5-1%；
Quality volumn concentration of the dextran-40 in the reagent is 0.5%；
Quality volumn concentration of the leucine in the reagent is 0.5-1%；
Volumn concentration of the poloxamer in the reagent is 0.05%.
In mentioned reagent,
The drug is albumen or other chemical moleculars；
The albumen is antigen or protease；Antigen protein includes the various eggs for having therapeutic effect or immune response stimulating
It is white.
And/or the protease is specially trypsase.
Another object of the present invention is to provide a kind of dry powder inhaler formulations.
Dry powder inhaler formulations provided by the invention obtain dry powder inhaler formulations for above-mentioned reagent is prepared as dry powder.
The aerodynamic qualities median diameter of above-mentioned dry powder inhaler formulations is 1-5 μm.
Or the preparation method is that atomizing freeze drying method；
Or the fog-spray nozzle that the atomizing freeze drying method uses is two fluid spray head, supply gas pressure 0.1-0.2MPa,
Feed flow rate is 4-8mL/min.
The particle size range of above-mentioned dry powder inhaler formulations is 1 μm -25 μm.
3rd purpose of the invention is to provide a kind of method for preparing above-mentioned dry powder inhaler formulations.
Method provided by the invention, includes the following steps:
1) above-mentioned reagent is prepared；
2) reagent is prepared into dry powder inhaler formulations using atomizing freeze drying method.
In the above method,
The fog-spray nozzle that the spray-freezing drying method uses is two fluid spray head, supply gas pressure 0.1-0.2MPa,
Feed flow rate is 4-8mL/min.
Above-mentioned dry powder inhaler formulations preparation human or animal's vaccine or treat human or animal's disease product in application be also
The scope of protection of the invention.
It is maintained at 1-5 μm commonly used in the drug aerodynamic size of sucking and can just there is the preferable lung that reaches to lead concurrently
Drug effect is waved, 5 μm of particulate matters below of diameter can enter the deep of respiratory tract, and 2.5 μm of diameter or so of particulate matter can be as deep as lung
It steeps and deposits, and comparatively, particle size is bigger, the active drug amount carried is also bigger, therefore 2.5 μm of left sides of diameter
Right particulate matter is preferably to select.Mass median diameter usually can preferably represent the distribution characteristics of particulate matter.The present invention
Novel dry powder suck preparation technology of preparing, its aerodynamic qualities median diameter of the solid dry powder particles prepared is in 1-
Between 5 μm, it can preferably be deposited on lung.
Invent a kind of novel to supplement existing microorganism infection related vaccines preparation or respiratory administration drug, inventor
Dry powder inhaler formulations technology of preparing, the dry powder inhaler formulations technology of preparing are used drug or antigen protein and immunologic adjuvant, several
The solution of kind excipient uses SFD technology after mixing, to mixed solution, prepares the suitable solid dry powder particles of partial size, close
Envelope saves；Before use, solid dry powder is packed into dry powder aerosol generator, acceptor occurs inhalable dry powder aerosol,
Drug is sucked respiratory tract and lung by breathing by human or animal.
Novel dry powder of the present invention sucks preparation, before use, being packed into appropriate solid powder according to recipient's difference
Dry powder aerosol occurs for suitable dry powder aerosol generator.Recipient is sucked dry powder by respiration, is exempted to reach
The purpose or effect of epidemic disease stimulation or disease treatment.
Novel dry powder of the present invention sucks preparation, and with solidapowder form storage and transport, when preservation guards against damp.
Formula is different, then the water content of solid dry powder and water absorbing capacity difference.The dry powder particles dispersibility prepared is preferably, loose more
Hole, soluble easily in water, easily aerosolized, aerodynamic size is smaller, is suitable for use as breathing vaccine or breathing drug.
This research is intended to prepare aerosolizable protein dry powder inhalant.Wherein, destination protein is wrapped in excipient base
In, the aerodynamic qualities median diameter of Foradil Aerolizer formoterol fumarate is between 1~5 μm, and particle is loose porous, and particle diameter distribution is more equal
It is even.The Foradil Aerolizer formoterol fumarate aerodynamic qualities median diameter that this research is finally prepared by atomizing freeze drying technology is in 2.5 μ
M or so can be deposited on target organ preferably to stimulate the immune response of body.
Its aerodynamic qualities median diameter of dry powder inhaler formulations prepared by the present invention, can be preferable at 2.5 μm or so
Be deposited on target organ to stimulate body immune response or play effect of drugs.Moreover, being solid dry powder after being successfully prepared
Shape particulate matter, water content is low, and wettability power is weak, dispersed (spherical particle, partial size are smaller), relative to liquid preparation dry powder system
Agent is more easily transported, and is conducive to save, and water-soluble is good, has preferable respiratory immune or respiratory administration prospect.
Detailed description of the invention
Fig. 1 is the powder morphology of the dry powder inhaler formulations of embodiment 1- example 5.
Fig. 2 is the particle diameter distribution of 50% particulate matter quality among the dry powder inhaler formulations of embodiment 1- example 5.
Fig. 3 be embodiment 1- example 5 dry powder inhaler formulations water suction percentage-time graph, determination condition be 25 DEG C, 60%
RH。
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Quality concentration expressed in percentage by volume is quality unless otherwise specified in following embodiments: volume g:ml.
To facilitate the understanding of the present invention, a more comprehensive description of the invention is given in the following sections with reference to the relevant attached drawings.In attached drawing
Give presently preferred embodiments of the present invention.But the invention can be realized in many different forms, however it is not limited to this paper institute
The embodiment of description.On the contrary, purpose of providing these embodiments is keeps the understanding to the disclosure more thorough
Comprehensively.
Trypsinogen (PanReac AppliChem, A4532,0001), mannitol (Sigma, M1902), inositol
(Sigma, I7508), trehalose (Sigma, T0167), leucine (Sigma, L8912), poloxamer (Sigma, P5556).
Embodiment 1 sucks preparation by the Novel dry powder of model proteins of trypsinogen
1, P1 sucks the preparation of preparation mixed solution
P1 sucking preparation mixed solution is made of each solute as shown in table 1 below and solvent (water), and the concentration of each solute
(no specified otherwise is then quality concentration expressed in percentage by volume, and quality: volume (g:ml) is concentration shown in table 1.
Table 1 is the solute that Novel dry powder sucks preparation P1
By P1 sucking preparation adjusting pH value to 7.2,0.2 μm of filter filtration sterilization is then used, it is spare.
The preparation method of P1 sucking preparation mixed solution:
The mother liquor of each solute is first configured, specifically: 0.5% destination protein aqueous solution, 10% inositol aqueous solution can be heated and be helped
Molten, 10% aqueous trehalose solution, can heat hydrotropy；2% leucine aqueous solution, can heat hydrotropy；10% mannitol can be heated and be helped
It is molten；Again by sample is prepared shown in table 1 after, adjust pH value to 7.2, then use 0.2 μm of filter filtration sterilization.
2, Novel dry powder sucks preparation P1
P1 sucking preparation mixed solution prepares dry powder using SFD technology, obtains Novel dry powder sucking preparation P1, and method is specific
It is as follows:
1, air compressor is opened to and is connected to two fluid spray heads (TSE), adjusting air pressure is about 0.15MPa, feed flow
Rate is 4-8mL/min.
2, the stainless-steel pan for filling about 3/4 volume liquid nitrogen and magnetic stirring apparatus rotor is placed on magnetic stirring apparatus, it is second-rate
Spray body head frame is above stainless-steel pan.Then the formula P1 sucking preparation mixed solution that 2h is pre-chilled in 0 DEG C is transferred to 20mL
Syringe is inserted into two fluid spray heads by syringe, starts to carry out spray chilling.
3, after spray chilling, the ice crystal state solid matter in stainless-steel pan is transferred to stainless steel together with a small amount of liquid nitrogen
In bottle, almost volatilizees to liquid nitrogen totally, stainless steel cylinder is transferred in freeze drier and is freeze-dried.
4, sample is taken out after being freeze-dried 36h, is then weighed, is covered, is sealed, is saved, Novel dry powder sucking system is obtained
Agent P1.
Embodiment 2 sucks preparation by the Novel dry powder of model proteins of trypsinogen
P2 sucking preparation mixed solution is made of each solute as shown in table 2 below and aqueous solvent, and the concentration of each solute is
Concentration shown in table 2.
Table 2 is the solute that Novel dry powder sucks preparation P2
Wherein, before preparing mixed solution, mother liquor is prepared in advance.Preparation program is referring to embodiment 1.
2, Novel dry powder sucks preparation P2
Preparation process: with embodiment 1, used mixed solution is that P2 sucks preparation mixed solution.
Embodiment 3 sucks preparation by the Novel dry powder of model proteins of trypsinogen
P3 sucking preparation mixed solution is made of each solute as shown in table 3 below and aqueous solvent, and the concentration of each solute is
Concentration shown in table 3.
Table 3 is the solute that Novel dry powder sucks preparation P3
Wherein, before preparing mixed solution, mother liquor is prepared in advance.Preparation program is referring to embodiment 1.
2, Novel dry powder sucks preparation P3
Preparation process: with embodiment 1, used mixed solution is that P3 sucks preparation mixed solution.
Embodiment 4 sucks preparation by the Novel dry powder of model proteins of trypsinogen
P4 sucking preparation mixed solution is made of each solute as shown in table 4 below and aqueous solvent, and the concentration of each solute is
Concentration shown in table 4.
Table 4 is the solute that Novel dry powder sucks preparation P4
Wherein, before preparing mixed solution, mother liquor is prepared in advance.Preparation program is referring to embodiment 1.
2, Novel dry powder sucks preparation P4
Preparation process: with embodiment 1, used mixed solution is that P4 sucks preparation mixed solution.
Embodiment 5 sucks preparation by the Novel dry powder of model proteins of trypsinogen
P5 sucking preparation mixed solution is made of each solute as shown in table 5 below and aqueous solvent, and the concentration of each solute is
Concentration shown in table 5.
Table 5 is the solute that Novel dry powder sucks preparation P5
Wherein, before preparing mixed solution, mother liquor is prepared in advance.Preparation program is referring to embodiment 1.
2, Novel dry powder sucks preparation P5
Preparation process: with embodiment 1, used mixed solution is that P5 sucks preparation mixed solution.
The examination criteria of embodiment 6, Novel dry powder sucking preparation
Three parts of Novel dry powders containing trypsinogen, which are respectively prepared, by 1-5 of embodiment of the present invention the method sucks preparation,
Measure following index:
One, measurement trypsinogen is the dry powder inhaler formulations freeze-drying front and back proenzyme specific activity of enzyme of model proteins, calculates enzyme
Specific activity of enzyme loss late in original
The protein dry powder proenzyme Rate activity of five different formulations is measured, the reagent and instrument of use are as follows: pancreas egg
White enzyme activity determination kit (Abcam, ab102531), trypsase (Amresco, 0785), trypsinogen (PanReac
AppliChem, A4532), climatic chamber (the permanent experimental instrument and equipment Co., Ltd of Shanghai one), boric acid (Chinese medicines group),
CaCl2·2H2The NaOH etc. of O (Sigma), 1M
Experimental method: prepared as follows before experiment:
(1) borate buffer: 0.1M boric acid+0.1M CaCl is prepared2, pH 8.0；
(2) it (is equivalent to SFD method using the original solution that borate buffer prepares P1-P5 respectively and prepares the spraying use before dry powder
Solution), and dissolve dry composition solution using borate buffer simultaneously, in the dry powder solution of original solution and preparation trypsinogen and
The final concentration of trypsase is respectively that the mass ratio of 0.5mg/mL and 0.01mg/mL, i.e. trypsinogen and trypsase are 50:1；
(3) by prepared each reaction solution, 25 DEG C of climatic chambers is placed in and are incubated for about 8h, are then carried out using borate buffer
After 400 times of dilutions, using trypsase vitality test kit, referring to trypsinogen vigor in its specification detection each sample.
(4) finally into each sample be added reaction mixture after, at once in 405nm at detection absorbance value, early period every
2min or so detects an absorbance value, and the later period is according to detection case appropriate adjustment detection time spacing.It is read according to microplate reader
Calculate enzyme activity.
Enzyme activity calculation method is as follows:
All data subtract zero hole, then draw p-NA standard curve.Δ A405nm is applied to standard curve to obtain
To the nmol of p-NA (data are generated between T1 and T2).
B is the p-NA (nmol) calculated from standard curve；T1 and T2 is for the first time and second reads (min)；V
It is to be added to the pretreated sample volume (mL) of reacting hole.
Unit definition: 25 DEG C, trypsase per minute cracks substrate, generates the p-NA of 1.0 μm of ol, is defined as tryptose
One unit (1U) of enzyme.
(5) pancreas egg in each sample can be calculated according to trypsase vigor in unit volume solution and trypsase quality
White original Rate activity, formula are as follows:
The specific activity of enzyme loss late of each formula is further calculated according to the Rate activity of trypsinogen in each sample:
As a result it see the table below shown in 6:
The freeze-drying front and back proenzyme specific activity of enzyme loss late of table 6 (mean+SD that three parts of repetitions prepare sample)
| Novel dry powder sucks preparation | Specific activity of enzyme loss late (%) |
| P1 | 10.4±2.6 |
| P2 | 18.2±2.6 |
| P3 | -0.5±1.6 |
| P4 | 16.1±11.9 |
| P5 | 24.8±10.3 |
The result shows that the proenzyme Rate activity in the embodiment 1-5 of freeze-drying front and back has different degrees of decline, wherein in sample P 3
Proenzyme Rate activity loss reduction.
Two, the aerodynamic size test of dry powder inhaler formulations
Test method:
Instrument equipment mainly has aerodynamic size spectrometer (APS-3321, Technical Sourcing Internation, the U.S.), small-sized biological gas
Colloidal sol sedimentation evaluation and test cabin (Beijing intelligent flourish and Science and Technology Ltd.), hand-held dry powder aerosol lung delivery apparatus (Beijing it is intelligent flourish and
Science and Technology Ltd.), mass concentration detector (8530, Technical Sourcing Internation, the U.S.).
Concrete operations process is as follows:
1, a small fan is put to single layer animal systemic exposure system bottom, then runs single layer animal systemic exposure system
System opens contamination mode and carries out system self-cleaning, while in single layer animal systemic exposure system side interface quality of connection concentration
Detector, the particle concentration inside real-time monitoring exposure system are dense to single layer animal whole body dynamic exposure system endoparticle object
Degree is down to 0.006mg/m3And when following, the contamination mode of single layer animal systemic exposure system is closed.
2, for dry powder particle object, contain pancreas using hand-held dry powder aerosol lung delivery apparatus generation embodiment 1-5
(each dry powder sample uses 5mL gas that 5mg occurs to the Novel dry powder sucking preparation P1-P5 dry powder aerosol of proproteinase every time
Sample) after, fan 30S is run, concentration inspection of improving quality then is connected at the connector of single layer animal systemic exposure system side
Instrument, aerodynamic size spectrometer and pressure detecting table are surveyed, after fan is out of service, runs aerodynamic size spectrometer, even
Continuous sampling 5min, while being changed using mass concentration detector monitoring single layer animal systemic exposure system endoparticle object concentration.
3, after to aerodynamic size spectrometer sampling, the data of instrument record is read, the sky of every group of sample is recorded
Aerodynamics mass median diameter.Summarizing to the reading of the sample for repeating preparation three times of each formula, (three parts of repetitions are made
Reading average value+standard deviation of standby sample).
It the results are shown in Table shown in 7:
Table 7 is the test (unit μm) of aerodynamic qualities median diameter
| Novel dry powder sucks preparation | Aerodynamic qualities median diameter |
| P1 | 3.593±0.245 |
| P2 | 2.670±0.547 |
| P3 | 2.857±0.307 |
| P4 | 2.030±0.287 |
| P5 | 3.057±0.549 |
The result shows that the aerodynamics of the Novel dry powder sucking preparation P1-P5 containing trypsinogen of embodiment 1-5
Partial size is not quite similar, but difference is not counting very big.Comparatively, the aerodynamic size of Novel dry powder sucking preparation P2 is for exhaling
It inhales most more particularly suitable.
Three, the moisture determination of dry powder inhaler formulations
With instrument thermogravimetric analyzer (DISCOVERY, TA company, the U.S.) detection embodiment 1-5 containing trypsinogen
The water content of Novel dry powder sucking preparation P1-P5.
The results are shown in Table 8,
8 moisture determination result (%) of table
---: water content is extremely low, and undetermined goes out data
The result shows that each dry powder sample its water content prepared in embodiment 1-5 is not quite similar, P3 sample moisture content phase
To highest, P2 and P4 sample moisture content be can be ignored.
Four, trypsinogen is the dry powder inhaler formulations scanning electron microscopic observation of model proteins
Instrument: scanning electron microscope (S-3400N, Hitachi, Japan)
Experiment flow: observing each sample using scanning electron microscope after film-making, and contains to embodiment 1-5
The random measurement wherein 150 respectively of the dry powder samples of three parts of the preparation P1-P5 repetition preparations of Novel dry powder sucking of trypsinogen
The partial size of a or so single particle records its absolute particle size, further progress statistical analysis.
It is directly observed with scanning electron microscope, as a result visible Fig. 1.
To in the Novel dry powder sucking preparation P1-P5 containing trypsinogen of the embodiment 1-5 of measurement particulate matter it is exhausted
Its mass median diameter partial size corresponding with intermediate 50% mass is calculated after summarizing to partial size, as a result visible Fig. 2, intermediate point in figure
For particulate matter quality median diameter, the corresponding partial size of upper and lower ends indicates the total particle weight of particulate matter Zhan during this partial size
50%.
The result shows that spherical shape, loose porous, quality is all presented in each dry powder sample prepared in embodiment 1-5 substantially
It is relatively light.
Five, the water suction percentage of dry powder inhaler formulations P1-P5-time graph measurement
Instrument: climatic chamber (Shanghai Yiheng Scientific Instruments Co., Ltd)
Experiment flow: climatic chamber temperature and humidity is set as 25 DEG C, electronic balance is placed in climatic chamber by 60%RH
In cabin, start to be tested after parameter stability.The batch cultur ware of one diameter 9cm is put into the balance into climatic chamber
On, after zeroing, Novel dry powder sucking preparation P1-P542 ± 10mg containing trypsinogen of embodiment 1-5 is weighed, balance
Leeward side and top glass baffle plate are opened or stay seam, and the door of climatic chamber is then shut off, and record a dry powder weight every 30min
Amount, records 6h in total.
It calculates each sample and increases percentage (%) relative to the weight at 0 time point at every point of time, draw water suction hundred
Divide ratio-time graph.
As a result see Fig. 3, it can be seen that each dry powder sample its water absorbing capacity prepared in embodiment 1-5 is different,
The water absorbing capacity of middle P3 is most strong, and the water absorbing capacity of P5 and P2 is relatively weak.
Can be used as vaccine after trypsinogen in the Foradil Aerolizer formoterol fumarate is changed to antigen protein, can be used for suck to
Medicine.Dry powder aerosol can occur to recipient (can be human or animal) by dry powder aerosol generator, recipient passes through
Respiration actively or passively sucks dry powder aerosol, evokes lung's immune response or healing disease purpose to reach.
Claims (10)
1. a kind of reagent for being used to prepare dry powder inhaler formulations, including solute,
The solute is made of drug, mannitol, freeze drying protectant and dispersing agent；
The freeze drying protectant is at least one of inositol, trehalose and dextran-40；
The dispersing agent is made of leucine and poloxamer.
2. reagent according to claim 1, it is characterised in that:
The drug, the mannitol, the freeze drying protectant, the leucine and the poloxamer proportion be 0.05-
0.15g:1g:1-1.5g:0.5-1g:0.05ml；
Or, the freeze drying protectant is inositol and trehalose, the mass ratio of the inositol and the trehalose is 1:1；
Or, the freeze drying protectant is inositol and dextran-40, the mass ratio of the inositol and the dextran-40 is
1:0.5。
3. reagent according to claim 1 or 2, it is characterised in that:
The reagent further includes solvent,
The reagent is made of the solute and the solvent；
Quality volumn concentration of the drug in the reagent is 0.05-0.15%；
Quality volumn concentration of the mannitol in the reagent is 1%；
The freeze-dried quality volumn concentration in the reagent is 1-1.5%；
Quality volumn concentration of the inositol in the reagent is 0.5-1%；
Quality volumn concentration of the trehalose in the reagent is 0.5-1%；
Quality volumn concentration of the dextran-40 in the reagent is 0.5%；
Quality volumn concentration of the leucine in the reagent is 0.5-1%；
Volumn concentration of the poloxamer in the reagent is 0.05%.
4. reagent according to claim 1 to 3, it is characterised in that:
The drug is albumen or other chemical moleculars；
The albumen is antigen or protease；
And/or the protease is specially trypsase.
5. a kind of dry powder inhaler formulations obtain dry powder sucking for the reagent any in claim 1-4 is prepared as dry powder
Preparation.
6. dry powder inhaler formulations according to claim 5, it is characterised in that: the aerodynamics of the dry powder inhaler formulations
Mass median diameter is 1-5 μm；
Or the preparation method is that atomizing freeze drying method；
Or the fog-spray nozzle that the atomizing freeze drying method uses is two fluid spray heads, supply gas pressure 0.1-0.2MPa, feed flow
Rate is 4-8mL/min.
7. dry powder inhaler formulations according to claim 5 or 6, it is characterised in that: the partial size of the dry powder inhaler formulations
Range is 1 μm -25 μm.
8. a kind of method for preparing any dry powder inhaler formulations in claim 5-7, includes the following steps:
1) any reagent in claim 1-4 is prepared；
2) reagent is prepared into dry powder inhaler formulations using atomizing freeze drying method.
9. according to the method described in claim 8, it is characterized by:
The fog-spray nozzle that the spray-freezing drying method uses is two fluid spray heads, supply gas pressure 0.1-0.2MPa, feed flow
Rate is 4-8mL/min.
10. any dry powder inhaler formulations are in preparation human or animal's vaccine or treatment human or animal's disease in claim 5-7
Application in product.
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| CN113908254A (en) * | 2021-10-19 | 2022-01-11 | 山西锦波生物医药股份有限公司 | Dry powder inhalant and preparation method and application thereof |
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| CN111346222A (en) * | 2020-03-30 | 2020-06-30 | 中国人民解放军军事科学院军事医学研究院 | Preparation method of plague attenuated live vaccine dry powder |
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Inventor after: Zhou Dongsheng Inventor after: Zhao Yuee Inventor after: Yu Xuedong Inventor after: Yin Zhe Inventor after: Xiong Xiaolu Inventor after: Jiao Jun Inventor after: Jin Aijun Inventor after: Hu Lingfei Inventor after: Yang Wenhui Inventor after: Jiao Zhouguang Inventor after: Sun Yansong Inventor after: Qiu Yefeng Inventor after: Gao Bo Inventor after: Fa Yunzhi Inventor after: Yang Huiying Inventor after: Feng Junxia Inventor before: Jiao Zhouguang Inventor before: Zhao Yuee Inventor before: Yu Xuedong Inventor before: Yin Zhe Inventor before: Xiong Xiaolu Inventor before: Jiao Jun Inventor before: Jin Aijun Inventor before: Hu Lingfei Inventor before: Yang Wenhui Inventor before: Zhou Dongsheng Inventor before: Sun Yansong Inventor before: Qiu Yefeng Inventor before: Gao Bo Inventor before: Fa Yunzhi Inventor before: Yang Huiying Inventor before: Feng Junxia |