CN108992669A - Composite yolk antibody composition, aerosol inhalation solution agent and the preparation process and application for preventing and treating respiratory tract infection - Google Patents
Composite yolk antibody composition, aerosol inhalation solution agent and the preparation process and application for preventing and treating respiratory tract infection Download PDFInfo
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- CN108992669A CN108992669A CN201810983399.8A CN201810983399A CN108992669A CN 108992669 A CN108992669 A CN 108992669A CN 201810983399 A CN201810983399 A CN 201810983399A CN 108992669 A CN108992669 A CN 108992669A
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Abstract
A kind of composite yolk antibody composition, aerosol inhalation solution agent and preparation process and application preventing and treating respiratory tract infection, is related to composite yolk antibody field.The composite yolk antibody composition of the embodiment of the present invention is to be immunized to obtain using the antigen of the common pathogen strain including respiratory tract infection, the common pathogen of respiratory tract infection includes staphylococcus aureus, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa, its safety can prevent and treat respiratory tract infection；The application of the composite yolk antibody composition of the embodiment of the present invention is to be used to prepare the preparation for preventing and treating respiratory tract infection；The aerosol inhalation solution agent of the embodiment of the present invention and its preparation process are the aerosol inhalation solution agent prepared using composite yolk antibody composition for preventing and treating respiratory tract infection, absorb fast, significant effect.
Description
Technical field
The present invention relates to composite yolk antibody fields, and in particular to a kind of composite yolk antibody for preventing and treating respiratory tract infection
Composition, aerosol inhalation solution agent and preparation process and application.
Background technique
Respiratory tract infection is divided into the infection of the upper respiratory tract, lower respiratory tract infection, bacterial pneumonia or mixed cell sexuality dye
Viral pneumonia, be the most common infectious diseases.The current respiratory tract infection for the treatment of both at home and abroad is mainly anti-using Cephalosporins
Raw element, quinolone antibiotics and other synthetic antibacterial drugs.To control progression of the disease, the antibiosis that do not stint with large dosage, high concentration
As a result element has greatly accelerated bacterial resistance sexual development, an antibiotics listing is only used only 2 or three years, and antibody-resistant bacterium can go out
It is existing, and developing an antibiotics then needs 10 years or so.In addition, the toxic side effect of antibiotic, also startling.Based on all kinds of anti-
Various drawbacks of rhzomorph can no longer meet the demand of prevention and treatment respiratory tract infection clinical application now.
Composite yolk antibody (Immunoglobulin of Yolk, IgY) belongs to IgG immunoglobulin like protein, is with a variety of causes
Germ immune health breeding egg-laying bird is allowed to generate a large amount of corresponding immunoglobulins, extracted to isolate and purify, and is made special
Property composite yolk immune antiboidy.Composite yolk immune antiboidy can be specifically bound with corresponding antigens, to inhibit or change
The state and activity for becoming the antigen promote the phagocytosis of segmented cell or macrophage to pathogenic bacteria by opsonic action.But
In many cases, since diversification is usually presented in the underlying diseases of patient and the severity and delay of the state of an illness, pathogenic bacteria,
Usually based on mixed infection, therefore the effect of the selection of pathogenic bacteria directly affects the specific composite yolk antibody of acquisition.Mesh
The preceding common pathogen strain for using respiratory tract infection as antigen be immunized special chicken made from specificity composite yolk antibody pair
The treatment of respiratory tract infection is not to have good therapeutic effect to the respiratory tract infection of all kinds there is also limitation.
Therefore, it is necessary to develop a kind of safety, and the special efficacy preparation of all respiratory tract infection can be prevented and treated.
Summary of the invention
The purpose of the present invention is to provide a kind of composite yolk antibody composition for preventing and treating respiratory tract infection and its preparation works
Skill, product safety can prevent and treat all respiratory tract infection.
The purpose of the present invention is to provide a kind of applications of composite yolk antibody composition for preventing and treating respiratory tract infection, are used for
Preparation prevents and treats the preparation of respiratory tract infection.
The purpose of the present invention is to provide a kind of aerosol inhalation solution agent and its preparation processes, using composite yolk antibody group
It closes object and prepares the aerosol inhalation solution agent for preventing and treating respiratory tract infection, absorb fast, significant effect.
The present invention solves its technical problem and adopts the following technical solutions to realize.
The present invention proposes a kind of composite yolk antibody composition for preventing and treating respiratory tract infection, is using including respiratory tract sense
The common pathogen strain of dye is obtained as antigen is immune, the common pathogen of respiratory tract infection include staphylococcus aureus,
Hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa.
Further, in a preferred embodiment of the present invention, staphylococcus aureus, hemolytic streptococcus, streptococcus pneumonia,
Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa mass ratio be 4-8:4-8:3-5:4-8:4-8:3-5.
Further, in a preferred embodiment of the present invention, the pathogenic bacteria of respiratory tract infection further include staphylococcus epidermis, gold
Staphylococcus aureus and the mass ratio of staphylococcus epidermis are 4-8:3-5.
The present invention proposes a kind of preparation process of the composite yolk antibody composition of above-mentioned prevention and treatment respiratory tract infection, packet
Include following steps:
Preparation comprising staphylococcus aureus, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii,
The complex antigen of Pseudomonas aeruginosa；
Reinforced immunological is carried out to chicken with complex antigen, obtains the immunity eggs of immune chicken；
The yolk of immunity eggs is extracted and obtains crude extract dry powder；
Crude extract dry powder is isolated and purified by ion-exchange chromatography and gel permeation chromatography, obtains composite yolk antibody group
Close object.
The present invention proposes a kind of application of the composite yolk antibody composition of above-mentioned prevention and treatment respiratory tract infection, composite yolk
Antibody compositions prepare the aerosol inhalation solution agent for preventing and treating respiratory tract infection.
The present invention proposes a kind of aerosol inhalation solution agent, includes: above-mentioned compound ovum in every 100ml aerosol inhalation solution agent
Yellow antibody compositions 0.5-1.5g, buffering to and isotonic regulator, surplus be solvent.
Further, in a preferred embodiment of the present invention, buffering to selected from citric acid-sodium citrate, Acetic acid-sodium acetate,
One of citrate-phosphate disodium hydrogen, sodium dihydrogen phosphate-disodium hydrogen phosphate, lactic acid-sodium lactate, tartaric acid-sodium tartrate.
Further, in a preferred embodiment of the present invention, isotonic regulator is selected from sodium chloride, potassium chloride, magnesium chloride, chlorination
One of calcium, glucose, xylitol, sorbierite are a variety of.
The present invention proposes a kind of preparation process of above-mentioned aerosol inhalation solution agent comprising following steps:
Composite yolk antibody composition is added in partial solvent, is stirred evenly；Buffering is added to, isotonic regulator,
Stirring is to dissolving, and supplementing solvent is to enough, refined filtration after filtering.
Further, in a preferred embodiment of the present invention, additionally it is possible to 0.3-1g Chinese medicine extract is added, Chinese medicine extract
It is specific the preparation method comprises the following steps:
Climbing groundsel, honeysuckle, radix scrophulariae, polygonum cuspidate after cleaning is extracted with water at least twice, combined extract through high speed from
Scheming centrifugation, obtains the first centrifugate；
At least once with ethanol solution refluxing extraction, extracting solution is centrifuged the dregs of a decoction after water is mentioned through supercentrifuge, obtains
Two centrifugates；
Merge the first centrifugate and the second centrifugate, obtains Chinese medicine extract.
Composite yolk antibody composition, aerosol inhalation solution agent and the preparation of the prevention and treatment respiratory tract infection of the embodiment of the present invention
Technique and the beneficial effect of application are: the composite yolk antibody composition of the embodiment of the present invention is using including respiratory tract infection
The antigen of common pathogen strain is immune to be obtained, and the common pathogen of respiratory tract infection includes staphylococcus aureus, hemolytic
Streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa, safety can prevent and treat all exhale
Inhale road infection；The application of the composite yolk antibody composition of the embodiment of the present invention is to be used to prepare prevention and treatment respiratory tract infection
Preparation；The aerosol inhalation solution agent of the embodiment of the present invention and its preparation process are used using the preparation of composite yolk antibody composition
In the aerosol inhalation solution agent for preventing and treating respiratory tract infection, medication is convenient, absorbs fast, significant effect.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
Composite yolk antibody composition to the prevention and treatment respiratory tract infection of the embodiment of the present invention, aerosol inhalation solution agent below
And preparation process is specifically described with application.
The embodiment of the present invention provides a kind of composite yolk antibody composition for preventing and treating respiratory tract infection, is using including exhaling
Special chicken is immunized for the antigen for inhaling the common pathogen strain of road infection and separating-purifying obtains, specifically using respiratory tract infection
Common pathogen strain prepares complex antigen, and complex antigen is carried out reinforced immunological to special chicken, from obtained immune chicken
The yolk of immunity eggs extracts to obtain.The common pathogen of respiratory tract infection include staphylococcus aureus, hemolytic streptococcus,
Streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa, wherein staphylococcus aureus, hemolytic streptococcus, lung
Scorching streptococcus, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa mass ratio be 4-8:4-8:3-5:4-8:4-8:3-5.
In some embodiments of the invention, the pathogenic bacteria of respiratory tract infection further include staphylococcus epidermis (deposit number
ATCC12228), the mass ratio of staphylococcus aureus and staphylococcus epidermis is 4-8:3-5.Composite yolk antibody composition is
For the compound antibody extract for resisting above-mentioned respiratory tract infection pathogenic bacteria, composite yolk antibody composition is to leading to respiratory tract infection
Multiple pathogenic microorganisms have the specific binding suppression effect of going out, even if can also play useful effect in the case where concentration is extremely low,
And only pathogen is killed in suppression, will not generate antibody-resistant bacterium, also not find toxic side effect to human body.
The embodiment of the present invention provides a kind of preparation process of composite yolk antibody composition for preventing and treating respiratory tract infection, packet
Include following steps:
S1, preparation include staphylococcus aureus, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Bao Man not lever
Bacterium, Pseudomonas aeruginosa respiratory tract infection pathogenic bacteria complex antigen.
It is specifically respectively that staphylococcus aureus, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Bao Man is motionless
Bacillus, Pseudomonas aeruginosa constant temperature incubation 20-24h under the conditions of 34-36 DEG C, then inactivate spare；The Staphylococcus aureus that will have been inactivated
Bacterium, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa are according to mass ratio 4-8:4-8:
3-5:4-8:4-8:3-5 is uniformly mixed so as to obtain strain mixt；It is molten that strain mixt is dissolved in neutrality, the PBS of 0.08-0.1mol/L
In liquid, the strain mixt solution that concentration is 1.2-2.2mg/mL is made；Freund is added in the medium volume of strain mixt solution
Adjuvant, high speed homogenization is to get complex antigen.
S2, reinforced immunological is carried out to special chicken with complex antigen, obtains the immunity eggs of immune chicken.
Specifically complex antigen being injected at the chest muscle and thigh of 20 weeks big chickens, 0.5-0.6ml is injected at every place respectively,
Then primary every the injection of reinforcing in 10 days, it is at least 3 times immune altogether.The Freund's adjuvant in complex antigen used when injection is for the first time
Incomplete Freund's adjuvant, the Freund's adjuvant later in used complex antigen is complete Freund's adjuvant, last time for the second time
The produced egg of chicken for passing through reinforced immunological is collected after injection.
The chicken that reinforced immunological is carried out in the present embodiment is in hy-line brown hen, Jing Hongji, Yi Sha Cold boiled chicken and Bai Lai boat laying hen
It is a kind of；In order to keep composite yolk antibody composition quality obtained more preferable, the chicken for carrying out reinforced immunological selects by the following method:
7th month after injecting for the first time, IgY therein is extracted respectively after the produced egg of these chickens is marked, it is made with ELISA method detection
The potency of the IgY obtained；Compare its potency again, IgY solution potency >=1:128 chicken can be wherein made by selecting, then with this batch of chicken
Produced egg hatches chicken kind, grows up to it to 20 weeks, as the chicken for carrying out reinforced immunological.
S3, the yolk of immunity eggs is extracted to acquisition crude extract dry powder.
Specifically immunity eggs are smashed with eggbeater, egg white is removed, yolk is stirred evenly, by the 4-6 of yolk volume
Distilled water is added again to dilute and be uniformly mixed, debugs pH to 5.0-6.0；It is subsequently cooled to 2-6 DEG C, stands 12-24h；After pass through
High-speed centrifuge is centrifugated 20-30 minutes；Separating obtained supernatant is added in ultrafilter and carries out ultrafiltration and is concentrated
10-20 times, the metabisulfite solution that mass concentration is 10%-13% is added in the slurry after concentration and is sufficiently stirred；Again through height
Fast centrifuge centrifuge separation, takes supernatant, removes delipidized protein, obtain slurry；It adds slurry into ultrafilter and crosses ultra micro film to filter
Degerming；Slurry after filtration sterilization is freeze-dried with freeze drier, crude extract dry powder is made.
S4, crude extract dry powder is subjected to ion-exchange chromatography and gel exchange column progress Gel filtration through ion exchange column
Analysis obtains composite yolk antibody composition to isolate and purify.
The embodiment of the present invention provides a kind of application of the composite yolk antibody composition of above-mentioned prevention and treatment respiratory tract infection, multiple
It closes Yolk antibody composition and prepares the aerosol inhalation solution agent for preventing and treating respiratory tract infection.
The embodiment of the present invention provides a kind of aerosol inhalation solution agent, includes: above-mentioned in every 100ml aerosol inhalation solution agent
Composite yolk antibody composition 0.5-1.5g, buffering to and isotonic regulator, surplus be solvent.Wherein, buffering is to selected from citron
Acid-sodium citrate, Acetic acid-sodium acetate, citrate-phosphate disodium hydrogen, sodium dihydrogen phosphate-disodium hydrogen phosphate, lactic acid-sodium lactate,
One of tartaric acid-sodium tartrate.Isotonic regulator is selected from sodium chloride, potassium chloride, magnesium chloride, calcium chloride, glucose, wood
One of sugar alcohol, sorbierite are a variety of.
It in some embodiments of the invention, can also include 0.3-1g Chinese medicine extract, Chinese medical extract is using packet
Include climbing groundsel, honeysuckle, radix scrophulariae, polygonum cuspidate Chinese herbal medicine extract to obtain.In each Chinese herbal medicine, climbing groundsel has clearing heat and detoxicating work
With curing mainly the infection of the upper respiratory tract；Honeysuckle has stronger restraint to various pathogens and infection of the upper respiratory tract Causative virus etc.；
Radix scrophulariae, polygonum cuspidate can enhance Abwehrkraft des Koepers.Chinese medicine extract, Neng Gouti is made in climbing groundsel, honeysuckle, radix scrophulariae, polygonum cuspidate extraction
The antibacterial effect of high product and the effect of strengthen immunity.
In some embodiments of the invention, raw material further includes 0.3-1g lavo-ofloxacin, and lavo-ofloxacin is for treating
Acute/chronic bronchitis, asthmatic bronchitis, bronchial asthma and chronic pulmonary inflammatory thick sputum are not easy expectoration or dry cough
Without phlegm.
The aerosol inhalation solution agent of the present embodiment uses at least one of Chinese medical extract, lavo-ofloxacin (Western medicine)
With composite yolk antibody composition act synergistically, can further expansion antimicrobial spectrum, thus achieve the purpose that treat respiratory tract infection.
The embodiment of the present invention provides a kind of preparation process of above-mentioned aerosol inhalation solution agent comprising following steps:
(1) composite yolk antibody composition is slowly added into the solvent of recipe quantity 50%-80%, room temperature is slowly stirred
It is even, it is spare.
(2) it weighs suitable water for injection and is cooled to 50-70 DEG C, be slowly added to buffering pair, stirring is cold to being completely dissolved
But spare to room temperature.
(3) it weighs suitable water for injection and is cooled to 50-70 DEG C, be slowly added to isotonic regulator, stirring is cold to dissolving
But spare to room temperature.
(4) above-mentioned (2) and (3) are added in (1), are stirred evenly, injection active carbon is added after being heated to 30-40 DEG C
0.01%-0.1% (w/v) is stirred for 15-30min, filters decarburization.
(5) solvent is added to full dose, is stirred evenly, is detected pH and content, adjusts pH between 5.0-8.0.The present embodiment
PH value is adjusted using pH adjusting agent, pH adjusting agent includes at least to be selected from citric acid, malic acid, tartaric acid, boric acid, sodium lactate, hydrogen
At least one of sodium oxide molybdena.
(6) above-mentioned medical fluid is filtered through 0.5-1.5 μm of titanium filter core, detects intermediate visible foreign matters and pH value, adjust control
PH is between 5.0-8.0.
(7) by 0.2-0.25 μm of miillpore filter refined filtration of above-mentioned medical fluid, and every 2ml is poured under A grades of laminar flows are protected
Sterile ampoule bottle in, and fill nitrogen, sealing.
(8) after lamp inspection is qualified, and visible foreign matters, pH value, aseptic condition and the antibody titer of ampoule bottle content are detected, it can
See foreign matter should meet regulation, pH value detection range between 5.0-8.0, sterile, antibody titer >=1:128 is to get qualified mist
Change solution for inhalation.
In some embodiments of the invention, 0.3-1g Chinese medicine extract, the specific system of Chinese medicine extract can also be added
Preparation Method are as follows:
Climbing groundsel, honeysuckle, radix scrophulariae, polygonum cuspidate after cleaning is extracted with water at least twice, combined extract through high speed from
Scheming centrifugation, obtains the first centrifugate；
At least once with ethanol solution refluxing extraction, extracting solution is centrifuged the dregs of a decoction after water is mentioned through supercentrifuge, obtains
Two centrifugates；
Merge the first centrifugate and the second centrifugate, obtains Chinese medicine extract.
Feature and performance of the invention are described in further detail with reference to embodiments.
Embodiment 1
The present embodiment provides a kind of composite yolk antibody composition, it is made according to following procedure:
Respectively by staphylococcus aureus, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, green
Purulence bacillus is inoculated into Luria-Bertani culture medium, and Luria-Bertani culture medium includes 2% by mass percentage
Tryptone, 1% yeast extract, 1% sodium chloride and 1.5% agar, under 35 DEG C of environment be used in three gas incubators
Then middle culture 22h by above-mentioned strain separating and is inoculated into broth bouillon, broth bouillon includes by mass percentage
1% beef extract, 2% tryptone, 0.5% sodium chloride, 1.5% agar, under 35 DEG C of environment constant temperature shake
It swings and cultivates 26h in incubator, the revolving speed of constant-temperature shaking incubator is 180rpm, is then inactivated spare；The golden yellow that will have been inactivated
Staphylococcus, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa are by weight 6:6:4:
6:6:4 is uniformly mixed so as to obtain strain mixt；Strain mixt is dissolved in obtained bacterial strain in neutral, 0.09mol/L PBS solution to mix
The strain mixt solution that concentration is 1.8mg/mL is made in fit solution；Freund is added in the medium volume of strain mixt solution
Adjuvant stirs evenly, and the full bacterium complex antigen of emulsus is made；Full bacterium complex antigen is put into high-speed homogenization machine, is crushed at a high speed even
Change, complex antigen is made.
Complex antigen obtained is injected at the chest muscle and thigh of 20 weeks big chickens, 0.5ml is injected at every place respectively, then
It is primary every the injection of reinforcing in 10 days, it is immunized four times altogether.Freund's adjuvant used is incomplete Freund's adjuvant when injecting for the first time, second
Freund's adjuvant used by secondary to the 4th time is complete Freund's adjuvant, and the chicken for passing through reinforced immunological is collected after last time is injected
Produced egg.
Immunity eggs are smashed with eggbeater, egg white is removed, yolk is stirred evenly, are steamed by 5 times of additions of yolk volume
Distilled water is diluted and is uniformly mixed, and debugs pH to 6.0；4 DEG C are subsequently cooled to, 20h is stood；After be centrifuged through high-speed centrifuge
Separation 25 minutes；Separating obtained supernatant is added in ultrafilter and carries out 15 times of ultrafiltration and concentration, is 11% by mass concentration
Metabisulfite solution be added to concentration after slurry in be sufficiently stirred；It is centrifugated again through supercentrifuge, takes supernatant, remove degreasing
Albumen obtains slurry；It adds slurry into ultrafilter and crosses ultra micro film with filtration sterilization；Slurry after filtration sterilization is dry with freezing
Dry machine is freeze-dried, and crude extract dry powder is made.
Crude extract dry powder is subjected to ion-exchange chromatography through ion exchange column and gel exchange column carries out gel permeation chromatography
To isolate and purify, composite yolk antibody composition is obtained.
Embodiment 2
The present embodiment provides a kind of composite yolk antibody composition, it is made according to following procedure:
Respectively by staphylococcus aureus, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, green
Purulence bacillus, staphylococcus epidermis are inoculated into Luria-Bertani culture medium, and Luria-Bertani culture medium presses quality percentage
Number meter includes 2% tryptone, 1% yeast extract, 1% sodium chloride and 1.5% agar, is used under 36 DEG C of environment
20h is cultivated in three gas incubators, then by above-mentioned strain separating and is inoculated into broth bouillon, broth bouillon presses quality
Percentage meter includes 1% beef extract, 2% tryptone, 0.5% sodium chloride, 1.5% agar, in 34 DEG C of rings
30h is cultivated under border in constant-temperature shaking incubator, the revolving speed of constant-temperature shaking incubator is 200rpm, is then inactivated spare；It will
The staphylococcus aureus of inactivation, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa,
Staphylococcus epidermis is uniformly mixed so as to obtain strain mixt by weight 8:8:3:4:4:3:4；By strain mixt be dissolved in it is neutral,
Strain mixt solution is made in the PBS solution of 0.1mol/L, the strain mixt solution that concentration is 2.2mg/mL is made；In bacterium
Strain mixing liquid solution medium body product is added Freund's adjuvant and stirs evenly, and the full bacterium complex antigen of emulsus is made；Full bacterium is compound anti-
Original is put into high-speed homogenization machine, and high speed is crushed and homogenized, and complex antigen is made.
Complex antigen obtained is injected at the chest muscle and thigh of 20 weeks big chickens, 0.6ml is injected at every place respectively, then
It is primary every the injection of reinforcing in 10 days, it is immunized four times altogether.Freund's adjuvant used is incomplete Freund's adjuvant when injecting for the first time, second
Freund's adjuvant used by secondary to the 4th time is complete Freund's adjuvant, and the chicken for passing through reinforced immunological is collected after last time is injected
Produced egg.
Immunity eggs are smashed with eggbeater, egg white is removed, yolk is stirred evenly, are steamed by 6 times of additions of yolk volume
Distilled water is diluted and is uniformly mixed, and debugs pH to 5.0；6 DEG C are subsequently cooled to, 15h is stood；After be centrifuged through high-speed centrifuge
Separation 20 minutes；Separating obtained supernatant is added in ultrafilter and carries out 10 times of ultrafiltration and concentration, is 10% by mass concentration
Metabisulfite solution be added to concentration after slurry in be sufficiently stirred；It is centrifugated again through supercentrifuge, takes supernatant, remove degreasing
Albumen obtains slurry；It adds slurry into ultrafilter and crosses ultra micro film with filtration sterilization；Slurry after filtration sterilization is dry with freezing
Dry machine is freeze-dried, and crude extract dry powder is made.
Crude extract dry powder is subjected to ion-exchange chromatography through ion exchange column and gel exchange column carries out gel permeation chromatography
To isolate and purify, composite yolk antibody composition is obtained.
Embodiment 3
The present embodiment provides a kind of aerosol inhalation solution agent, it is made according to following preparation process:
(1) the composite yolk antibody composition in 1g embodiment 1 is slowly added into the water for injection that 80ml is cooled to room temperature
In, it is slowly stirred uniformly, it is spare.
(2) it weighs suitable water for injection and is cooled to 60 DEG C, be slowly added to citric acid-sodium citrate, stirring is to complete
Dissolution, is cooled to room temperature, spare.
(3) it weighs suitable water for injection and is cooled to 60 DEG C, be slowly added to 2g sodium chloride, stirring is cooled to dissolving
Room temperature, it is spare.
(4) above-mentioned (2) and (3) are added in (1), are stirred evenly, injection active carbon is added after being heated to 35 DEG C
0.05%, it is stirred for 20min, filters decarburization.
(5) mending and injecting water to total volume is 100ml, stirs evenly, detects pH and content, and adjusting pH is 5.5.
(6) above-mentioned medical fluid is filtered through 1.0 μm of titanium filter cores, detects intermediate visible foreign matters and pH value, adjusting control pH is
5.5。
(7) by 0.22 μm of miillpore filter refined filtration of above-mentioned medical fluid, and it is poured under A grades of laminar flows are protected the nothing of every 2ml
In bacterium ampoule bottle, and fill nitrogen, sealing.
(8) after lamp inspection is qualified, and visible foreign matters, pH value, aseptic condition and the antibody titer of ampoule bottle content are detected, it can
See foreign matter and should meet regulation, pH value meets regulation, sterile, antibody titer >=1:128 is to get qualified aerosol inhalation solution agent.
Embodiment 4
The present embodiment provides a kind of aerosol inhalation solution agent, it is made according to following preparation process:
(1) by 1g embodiment 1 composite yolk antibody composition, 0.5g Chinese medicine extract (by climbing groundsel, honeysuckle,
Radix scrophulariae, polygonum cuspidate extract to obtain) it is slowly added into the water for injection that 50ml is cooled to room temperature, it is slowly stirred uniformly, it is spare.
(2) it weighs suitable water for injection and is cooled to 70 DEG C, be slowly added to citrate-phosphate disodium hydrogen, stirring is to complete
Fully dissolved is cooled to room temperature, spare.
(3) it weighs suitable water for injection and is cooled to 70 DEG C, be slowly added to total 2g magnesium chloride and glucose, stirring is to molten
Solution, is cooled to room temperature, spare.
(4) above-mentioned (2) and (3) are added in (1), are stirred evenly, injection active carbon is added after being heated to 40 DEG C
0.01%, it is stirred for 30min, filters decarburization.
(5) mending and injecting water to total volume is 100ml, stirs evenly, detects pH and content, and adjusting pH is 6.0.
(6) above-mentioned medical fluid is filtered through 1.0 μm of titanium filter cores, detects intermediate visible foreign matters and pH value, adjusting control pH is
6.0。
(7) by 0.22 μm of miillpore filter refined filtration of above-mentioned medical fluid, and it is poured under A grades of laminar flows are protected the nothing of every 2ml
In bacterium ampoule bottle, and fill nitrogen, sealing.
(8) after lamp inspection is qualified, and visible foreign matters, pH value, aseptic condition and the antibody titer of ampoule bottle content are detected, it can
See foreign matter and should meet regulation, pH value meets regulation, sterile, antibody titer >=1:128 is to get qualified aerosol inhalation solution agent.
Embodiment 5
The present embodiment provides a kind of aerosol inhalation solution agent, it is made according to following preparation process:
(1) composite yolk antibody composition, the 0.5g lavo-ofloxacin in 1g embodiment 2 are slowly added into 70ml and be cooled to
In the water for injection of room temperature, it is slowly stirred uniformly, it is spare.
(2) weigh suitable water for injection and be cooled to 60 DEG C, be slowly added to lactic acid-sodium lactate, stirring to being completely dissolved,
It is cooled to room temperature, it is spare.
(3) it weighs suitable water for injection and is cooled to 60 DEG C, be slowly added to total 2g potassium chloride and sorbierite, stirring is to molten
Solution, is cooled to room temperature, spare.
(4) above-mentioned (2) and (3) are added in (1), are stirred evenly, injection active carbon is added after being heated to 40 DEG C
0.1%, it is stirred for 15min, filters decarburization.
(5) mending and injecting water to total volume is 100ml, stirs evenly, detects pH and content, and adjusting pH is 5.0.
(6) above-mentioned medical fluid is filtered through 1.0 μm of titanium filter cores, detects intermediate visible foreign matters and pH value, adjusting control pH is
5.0。
(7) by 0.22 μm of miillpore filter refined filtration of above-mentioned medical fluid, and it is poured under A grades of laminar flows are protected the nothing of every 2ml
In bacterium ampoule bottle, and fill nitrogen, sealing.
(8) after lamp inspection is qualified, and visible foreign matters, pH value, aseptic condition and the antibody titer of ampoule bottle content are detected, it can
See foreign matter and should meet regulation, pH value meets regulation, sterile, antibody titer >=1:128 is to get qualified aerosol inhalation solution agent.
Embodiment 6
The present embodiment provides a kind of aerosol inhalation solution agent, it is made according to following preparation process:
(1) by 1g embodiment 2 composite yolk antibody composition, 0.3g Chinese medicine extract (by climbing groundsel, honeysuckle,
Radix scrophulariae, polygonum cuspidate extract to obtain), 0.3g lavo-ofloxacin be slowly added into the water for injection that 50ml is cooled to room temperature, be slowly stirred
Uniformly, spare.
(2) it weighs suitable water for injection and is cooled to 70 DEG C, be slowly added to citrate-phosphate disodium hydrogen, stirring is to complete
Fully dissolved is cooled to room temperature, spare.
(3) it weighs suitable water for injection and is cooled to 70 DEG C, be slowly added to total 2g magnesium chloride and glucose, stirring is to molten
Solution, is cooled to room temperature, spare.
(4) above-mentioned (2) and (3) are added in (1), are stirred evenly, injection active carbon is added after being heated to 40 DEG C
0.01%, it is stirred for 30min, filters decarburization.
(5) mending and injecting water to total volume is 100ml, stirs evenly, detects pH and content, and adjusting pH is 5.5.
(6) above-mentioned medical fluid is filtered through 1.0 μm of titanium filter cores, detects intermediate visible foreign matters and pH value, adjusting control pH is
5.5。
(7) by 0.22 μm of miillpore filter refined filtration of above-mentioned medical fluid, and it is poured under A grades of laminar flows are protected the nothing of every 2ml
In bacterium ampoule bottle, and fill nitrogen, sealing.
(8) after lamp inspection is qualified, and visible foreign matters, pH value, aseptic condition and the antibody titer of ampoule bottle content are detected, it can
See foreign matter and should meet regulation, pH value meets regulation, sterile, antibody titer >=1:128 is to get qualified aerosol inhalation solution agent.
It is investigated by therapeutic effect of the following clinical test to the aerosol inhalation solution agent of the embodiment of the present invention:
One, selection suffers from volunteer 50 of the infection of the upper respiratory tract, is controlled using the aerosol inhalation solution agent in embodiment 3
It treats, aerosol inhalation solution agent is specifically carried out to volunteer by atomization inspiration treatment using atomizer, atomization 2 times daily, every time
4mL, the various symptoms situations of change such as observation volunteer sneezing, nasal obstruction, stream clear water nasal mucus.
After volunteer's medication 5-7 days, the symptom of 45 volunteers disappears substantially, the symptom of remaining 5 volunteer mitigates；From
After hope person's medication 10 days, the symptom of all volunteers disappears.
Thus illustrate, the aerosol inhalation solution agent as made from the composite yolk antibody composition of the embodiment of the present invention is to above exhaling
The therapeutic effect for inhaling road infection is significant.
Two, selection suffers from volunteer 50 of lower respiratory tract infection, is controlled using the aerosol inhalation solution agent in embodiment 4
It treats, the aerosol inhalation solution agent is specifically carried out to volunteer by atomization inspiration treatment using atomizer, atomization 2 times daily, often
Secondary 4mL, the various symptoms situation of change such as observation volunteer's sneezing, rhinorrhea, pharyngalgia, out of strength, low-heat.
After volunteer's medication 5-7 days, the symptom of 46 volunteers disappears substantially, the symptom of remaining 4 volunteer mitigates；From
After hope person's medication 10 days, the symptom of all volunteers disappears.
Thus illustrate, the aerosol inhalation solution agent as made from the composite yolk antibody composition of the embodiment of the present invention is exhaled under
The therapeutic effect for inhaling road infection is significant.
Three, selection suffers from volunteer 50 of bacterial pneumonia, is treated using the aerosol inhalation solution agent in embodiment 5,
The aerosol inhalation solution agent is specifically carried out to volunteer by atomization inspiration treatment using atomizer, atomization 2 times daily, every time
4mL, the various symptoms situations of change such as observation volunteer's cough, expectoration (having purulent sputum or bloody sputum), pectoralgia.
After volunteer's medication 5-7 days, the symptom of 45 volunteers disappears substantially, the symptom of remaining 5 volunteer mitigates；From
After hope person's medication 10 days, the symptom of 49 volunteers disappears, the symptom of remaining 1 volunteer disappears substantially.
Thus illustrate, aerosol inhalation solution agent is to bacterium as made from the composite yolk antibody composition of the embodiment of the present invention
The therapeutic effect of property pneumonia is significant.
Four, volunteer 50 of viral pneumonia of the selection with mixed cell sexuality dye, in addition to using same virus
Outside drug therapy, also treated using the aerosol inhalation solution agent in embodiment 6, it is specifically using atomizer that the Neulized inhalation is molten
Liquor carries out atomization inspiration treatment, daily atomization 2 times, each 4mL, observation volunteer fever, headache, whole body acid to volunteer
Bitterly, the various symptoms situation of change such as dry cough and lung infiltration.
After volunteer's medication 5-7 days, the symptom of 44 volunteers disappears substantially, the symptom of remaining 6 volunteer mitigates；From
After hope person's medication 10 days, the symptom of 48 volunteers disappears, the symptom of remaining 2 volunteer disappears substantially.
Thus illustrate, aerosol inhalation solution agent is to mixing as made from the composite yolk antibody composition of the embodiment of the present invention
The therapeutic effect of the viral pneumonia of bacterial infection is significant.
In addition, the curative effect comparative approach and result of the treatment respiratory tract infection of embodiment 3-6 are as follows:
The Patients With Respiratory Tract Infection of the not yet medication of selection 80, and clinical symptoms: having yellow phlegm after fever 2 days and coughing has
2 days, no asthma medical history, patient age range 3-6 years old, male 40, female 40.Criterion of therapeutical effect is as follows:
Cure: without cough, without yellow phlegm, spirit it is good；
Effective: cough number significantly reduces, improves without yellow phlegm, spirit；
Invalid: cough number increases, yellow phlegm increases, spirit is not good enough.
80 Patients With Respiratory Tract Infections are divided into 4 groups of test groups, every 20 people of experimental group, every group of male, each 10 people of female, test group
1-4 is corresponding to use embodiment 3-6 to patient's atomization inspiration treatment.Treatment method is: in patient and its accompanying family members' informed consent
Under, using embodiment 3-6 to patient's atomization inspiration treatment, daily atomization medication 2 times, each medication 4ml.Statistics medication 5 days
Each group therapeutic effect after with 10 days, as shown in table 1.
The curative effect of the treatment respiratory tract infection of 1 embodiment 3-6 of table compares
As seen from the above table, the aerosol inhalation solution agent as made from the anti-various pathogens composition of the embodiment of the present invention is to hair
Treatment of cough effective percentage after burning is significant in efficacy up to 100% or more.
In conclusion the composite yolk antibody composition and its preparation process of the prevention and treatment respiratory tract infection of the embodiment of the present invention
Safety, can prevent and treat all respiratory tract infection；The application of the composite yolk antibody composition of the embodiment of the present invention is to use
The preparation of respiratory tract infection is prevented and treated in preparation；The aerosol inhalation solution agent of the embodiment of the present invention and its preparation process are to adopt
The aerosol inhalation solution agent for preventing and treating respiratory tract infection is prepared with composite yolk antibody composition, medication is convenient, inhales
Receive fast, significant effect.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiments of the present invention, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (10)
1. a kind of composite yolk antibody composition for preventing and treating respiratory tract infection, which is characterized in that it is using including respiratory tract sense
The antigen of the common pathogen strain of dye is immune to be obtained, and the common pathogen of the respiratory tract infection includes Staphylococcus aureus
Bacterium, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, Pseudomonas aeruginosa.
2. the composite yolk antibody composition of prevention and treatment respiratory tract infection according to claim 1, which is characterized in that the gold
Staphylococcus aureus, the hemolytic streptococcus, the streptococcus pneumonia, the Escherichia coli, the Acinetobacter bauamnnii, institute
The mass ratio for stating Pseudomonas aeruginosa is 4-8:4-8:3-5:4-8:4-8:3-5.
3. the composite yolk antibody composition of prevention and treatment respiratory tract infection according to claim 1, which is characterized in that respiratory tract
The pathogenic bacteria of infection further include staphylococcus epidermis, and the mass ratio of the staphylococcus aureus and the staphylococcus epidermis is
4-8:3-5。
4. a kind of preparation process of the composite yolk antibody composition of prevention and treatment respiratory tract infection as described in claim 1, special
Sign is comprising following steps:
Preparation includes staphylococcus aureus, hemolytic streptococcus, streptococcus pneumonia, Escherichia coli, Acinetobacter bauamnnii, green pus
The complex antigen of bacillus；
Reinforced immunological is carried out to chicken with the complex antigen, obtains the immunity eggs of immune chicken；
The yolk of the immunity eggs is extracted and obtains crude extract dry powder；
The crude extract dry powder is isolated and purified by ion-exchange chromatography and gel permeation chromatography, obtains composite yolk antibody group
Close object.
5. a kind of application of the composite yolk antibody composition of prevention and treatment respiratory tract infection as described in claim 1, feature exist
In the composite yolk antibody composition prepares the aerosol inhalation solution agent for preventing and treating respiratory tract infection.
6. a kind of aerosol inhalation solution agent, which is characterized in that include: such as claim 1 institute in every 100ml aerosol inhalation solution agent
The composite yolk antibody composition 0.5-1.5g stated, buffering to and isotonic regulator, surplus be solvent.
7. aerosol inhalation solution agent according to claim 6, which is characterized in that the buffering is to selected from citric acid-citron
Sour sodium, Acetic acid-sodium acetate, citrate-phosphate disodium hydrogen, sodium dihydrogen phosphate-disodium hydrogen phosphate, lactic acid-sodium lactate, tartaric acid-
One of sodium tartrate.
8. aerosol inhalation solution agent according to claim 6, which is characterized in that the isotonic regulator be selected from sodium chloride,
One of potassium chloride, magnesium chloride, calcium chloride, glucose, xylitol, sorbierite are a variety of.
9. a kind of preparation process of aerosol inhalation solution agent as claimed in claim 6, which is characterized in that it includes following step
It is rapid:
Composite yolk antibody composition is added in partial solvent, is stirred evenly；Buffering is added to, isotonic regulator, stirring
To dissolution, supplementing solvent is to enough, refined filtration after filtering.
10. the preparation process of aerosol inhalation solution agent according to claim 9, which is characterized in that 0.3- can also be added
1g Chinese medicine extract, the Chinese medicine extract specific the preparation method comprises the following steps:
Climbing groundsel, honeysuckle, radix scrophulariae, polygonum cuspidate after cleaning is extracted with water at least twice, combined extract is through supercentrifuge
Centrifugation, obtains the first centrifugate；
At least once with ethanol solution refluxing extraction, extracting solution is centrifuged the dregs of a decoction after water is mentioned through supercentrifuge, obtain second from
Heart liquid；
Merge the first centrifugate and the second centrifugate, obtains Chinese medicine extract.
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