WO2005040218A1

WO2005040218A1 - METHOD FOR PREPARING COMPOSITE IgY SPECIFIC AGAINST PERIODONTAL DISEASE AND COMPOSITE PREPARATION THEREOF

Info

Publication number: WO2005040218A1
Application number: PCT/CN2004/000222
Authority: WO
Inventors: Sing Paau; Rongjian Yang; Tongsen Li
Original assignee: Jason Medical Group (Far East) Limited
Priority date: 2003-08-15
Filing date: 2004-03-19
Publication date: 2005-05-06
Also published as: CN1482141A

Abstract

The present invention discloses a IgY specific against periodontal disease and its preparation method and preparation. The, preparation method includes the following steps: screening major pathogens causing periodontal disease, preparing single or composite pathogenic antigens individually on the basis of screening,classifying and combining each pathogen, preparing immunized eggs, preparing crude extract of IgY specific agaínst each pathogen causing periodontal disease, and purifying the said crude extraet. The various IgY preparation specific against periodontal disease has lasting effect, safety and convenience for preventing and treating periodontal disease.

Description

Method for preparing periodontal disease-specific compound Y and combination preparation thereof TECHNICAL FIELD

The invention relates to an anti-periodontal disease specific composite IgY and a preparation method thereof, in particular to a specific composite IgY for preventing and treating periodontal disease (gingivitis, periodontitis, etc.), a preparation method and a combined preparation thereof. Background technique

Periodontal disease is one of the most common oral diseases in humans. Because periodontal disease does not cause pain and large dysfunction in the early stage, patients are often easy to ignore until gum bleeding, soft pain, swelling and inflammation, pus loosening, slot bone Lesions before going to the doctor for treatment. Therefore, the incidence of periodontal disease is extremely high. According to statistics from the WHO survey, the prevalence of gingivitis in children and adolescents is 70-90%, and it increases with each passing year. The incidence of periodontal disease in the elderly reaches 80-90%. According to one source, the incidence rate in adults is as high as 95%. In terms of etiology, in addition to a small number of endocrine drug genetic and psychological factors, scholars in the past 50 years have recognized that all types of periodontal disease are caused by dental plaque infection and it has a higher prevalence than dental caries. Up to now, countries around the world do not have effective drug prevention and treatment, which is extremely harmful to human health.

At present, countries around the world use chemical drugs to treat or inhibit plaque bacteria. There have been basically no major changes in the past 100 years. There are a large number of research reports that have tested various drugs. Currently, the most commonly used drugs in the clinic are nitrotazoles, such as metronidazole, tinidazole, and diguanidine, such as hydrochloric acid. Bitain, chlorhexidine acetate, chlorhexidine gluconate, etc. These are broad-spectrum antibacterials. The biggest problem is that long-term use of broad-spectrum bacteriostatic drugs will produce drug-resistant strains and double infections and other side effects. In recent years, some enzyme preparations have been marketed, such as lysozyme, glucose oxidase, glucanase, SOD.H ₂ Although enzymes such as ₀₂ and RNase can reduce plaque formation, they have poor stability, are easily degraded and inactivated, and are difficult problems in the stability of oral medications.

The currently used fluoride treatment drugs (fluorine-containing mouthwashes, etc.) are due to factors such as "multi-fluorine uptake" and "high-fluorine regionality" of the human body and fluorinated water sources; treatment with fluoride will cause the harm of excessive fluoride Fluoride, fluorosis, and even carcinogenesis; treatment of periodontal disease with nitrimidazole, metronidazole, tinidazole, not only has extremely bitter taste, metal taste in the mouth, but prolonged use can cause liver dysfunction to accumulate poisoning and sodium Retention, induced candida albic stomatitis, peripheral neuritis, leukopenia, pregnancy risk, disulfiram reaction, allergies and other side effects; chlorhexidine is currently the recommended oral medication, which has a relatively good effect on dental caries and periodontal disease Good efficacy, but still cannot escape the disadvantages of broad-spectrum chemical drugs. Repeated use can cause mucosal gum allergy, dryness, swelling, and cracking. The residue of chloroaniline in its synthesis is a potential carcinogen. For this reason, the US FDA and the UK BP is extremely restrictive. In the treatment of rinsing and rinsing with this product, fixed eruption of hands and feet and mucosal damage often occur.

Therefore, a natural and safe method that can kill periodontal disease bacteria without toxic and side effects, and has a long-term effect The new medicine is very necessary. Anti-human periodontal disease-specific multiple yolk antibodies

(Immunoglobulin of Yolk, IgY) is an IgG immunoglobulin, which is used to immunize healthy egg-laying poultry with a variety of pathogens that cause periodontal disease, so that it produces a large number of corresponding immunoglobulins, which are extracted, separated, and purified to obtain specificity. Complex yolk antibody. These composite IgYs have specific binding inhibitory effects on a variety of pathogens that cause periodontal disease, and even at very low concentrations, they can also have an effective effect, and only inhibit pathogenic bacteria, but have no effect on oral symbiotic bacteria. It will never produce drug-resistant strains and double infections, and it has no toxic side effects of chemicals. Technical content

The purpose of the present invention is to provide a method for preparing specific compound IgY that directly inhibits multiple pathogenic microorganisms of periodontal disease and a preparation thereof; the object of the present invention is also to provide a specific compound that can inhibit periodontal pathogenic microorganisms IgY is a natural safe and effective preparation for the prevention and treatment of periodontal disease.

The object of the present invention is achieved by the following technical solutions −

(1) Screening the pathogenic microorganisms that cause periodontal disease:

According to the infection microecology literature and clinical stomatology epidemiological data, 31 pathogenic bacteria in periodontal plaque were isolated and screened according to the number and probability of their occurrence, and classified according to the "antigenic cross" of these most common pathogens. The combination finally determined the following 8 main pathogenic bacteria:

Porphyromonas gingivalis (Porp yromonas gingiValis)

Fusobacterium nucleatum

Actinobacillus actinomycetem comitans Actinomgces Viscosus

Bacteroides forsythus

Treponemas denticola

Bacteroides intermedius

Bacteroides gingivalis

(2) Prepare single or composite periodontal pathogen antigens on the basis of screening and classification and combination of these diseases and organisms: According to the culture characteristics and optimal culture conditions of these pathogenic microorganisms, select appropriate media and conduct large numbers of strains Multiply and expand, then harvest the culture for purification, strain counting, dilution and packing;

Preparation of composite bacterial antigen: The above-mentioned cultured and purified single antigen is referenced to the probability and proportion of periodontal disease plaque, and mixed as follows-Porphyromonas gingivalis 20-5%

, Fusobacterium nucleus 20-5% Actinomyces actinomycetes 20-5%

Actinomyces viscous 15-10%

Forsythia bacillus 10-20%

Tartar Treponema pallidum 5-15%

Intermediate Bacteria 5-20%

Gingival bacteria 5-20%

Add a certain amount of each single or compound periodontal pathogenic bacteria to Freund's adjuvant, then put it into a high-speed homogenizer to mix at 10000-30000r / min to make a uniform W / 0 emulsion to make a single Complex Periodontal Pathogen Antigen.

(3) Preparation of immune eggs

Select egg-laying SPF healthy hens with high immune response ability, and inject single or compound periodontal pathogen antigens into the chest muscles or subcutaneously at multiple points. After the first injection, the interval is 7 days. The second dose was injected at the same dose and method. The third dose was injected at the same dose for another 7 days after the second injection. After 20 days from the first injection, the high titer was selected based on the titer of the compound IgY in serum and egg yolk. Immune to eggs. Immune eggs are numbered for different antigens.

The hens can be laying hens or laying ducks or laying geese or laying turkeys or laying ostriches or laying pheasants.

(4) Preparation of anti-periodontal pathogen-specific composite IgY crude product

Soak or wipe the eggs produced by hens immunized with different antigens separately with disinfectant, rinse them with sterile pure water and dry them, then place them in an eggbeater, break the shells, filter the egg whites with a yolk sieve, and save Egg yolk, add 5-8 times distilled water to dilute and mix, adjust the pH to 5.5-6.0 with 1 mol NaOH or HCL solution, and let it stand overnight at 4-6 ° C. Centrifuge the diluted solution at 1000-3000 r / min for 20 minutes. clear concentrated 5-10 million molecular weight cutoff ultrafiltration membrane D _a 15-20 fold concentrated solution was dried by lyophilization, freeze-dried powder to a single antigen IgY of 1-10: 1-10: 1-10: 1-10 The ratio of 1-10: 1-10: 1-10: 1-10 is mixed to obtain the crude periodontal disease-specific composite IgY.

(5) Preparation of anti-periodontal pathogen-specific composite IgY pure product

The company's published technology (patent application number 00101270.3, patent publication number 1307061) was passed through an ion exchange resin column and a gel exchange column, respectively, and the crude periodontal disease-resistant composite IgY prepared according to the above process was purified to obtain pure Compound IgY.

The SDS-PAGE electrophoresis method was used to detect the above crude IgY, and the composite IgY content was measured to be above 50%.

(6) The titer of complex IgY biological activity measured by enzyme-linked immunoassay (ELISA) was 128-512.

By combining 0.02-80% of this specific IgY with 99.98-20% of medicinal excipients, it can be made into oral sprays, mouthwashes, lozenges, stick-shaped treatments or toothpastes that are acceptable in clinical dentistry. The above dosage form of the invention is directly applied to oral periodontal local prevention and treatment of periodontal disease, which has good curative effect and quick effect, and solves the disadvantages of using various chemotherapeutic drugs for oral intolerance, irritability, drug resistance and side effects, and for oral cavity. Symbiotic bacteria have no effect. Experimental Example 1: The pathogenic bacteria were 2 to 3.5 billion, of which Porphyromonas gingivalis was 20 ° /. 20% Clostridium nucleus, 20% actinomycete actinomycetes, 15% sticky actinomycetes, 10% forsythia, F. tartarii, 5% intermediate bacteria, 5% gingival bacteria, Make 0.5ml of bacterial solution, add 0.5ml Freund's adjuvant and mix thoroughly to prepare a complex antigen. Use it to immunize laying hens, pick eggs to prepare their specific complex IgY, and then use gingival porphyrins Bacteria, Clostridium sclerotiorum, actinomycete actinomycetes, Actinomyces viscosus, Pseudomonas fusiformis, Treponema pallidum, Intermediate bacillus, and Gingival bacilli for the detection of antigens by "ELISA" method. Periodontal disease-specific complex IgY antibody binding titer, the results are as follows:

From the above test results, it is shown that the specific complex IgY prepared by the immunized female poultry with the complex antigen prepared by the present invention has an ideal broad spectrum, and has a strong inhibitory effect on the immunized bacteria (antibody titer 1: 128 -512). Experimental Example 2: The method is the same as Experimental Example 1. The proportion of antigens for injection was changed to 20% gingival bacillus, 20% intermediate bacillus, 20% fusiform bacterium, 15% tartar Treponema pallidum, 10% viscous actinomycetes, 5% actinomycetes actinomycetes, nucleated When Clostridium 5% and Porphyromonas gingivalis 5%, the antibody binding titer results are as follows:

IgY detection antigen antibody binding titer

Gingival Bacterium 1: 512 Anti-Gingival Bacilli, Intermediate Bacterium, Intermediate Bacterium 1: 512 Forsythia bacillus, tartar Treponema pallidum, ffi-Nanpo 1: 512

Actinomyces viscous, Symbiotic actinomyces, Treponema pallidum 1: 512

Actinomyces, Clostridium nucleus, Porphyrin gingivalis Actinomyces viscosus 1: 256

Simmons-specific complex IgY actinomycetes actinomyces 1: 128

Fusobacterium nucleus 1: 128

Porphyromonas gingivalis 1: 128 The ratio of antigens can be seen from the above table. The specific complex IgY antibody binding titer extracted after intensive immunization also changes accordingly, and the immune response is very good. Experimental Example 3: The composite antigen prepared in Experimental Example 1 was used to immunize laying hens, laying hens, laying geese, laying turkeys, and laying ostriches. Then, the method of the present invention is used to prepare: (1) anti-periodontal pathogen-specific chicken IgY, (2) anti-periodontal pathogen-specific duck IgY, (3) anti-periodontal pathogen Specific goose IgY, (4) anti-periodontal pathogen-specific turkey IgY, (5) anti-periodontal pathogen-specific ostrich IgY. Then, using the "ELISA method" (Enzyme-linked immunosorbent assay), P. gingivalis, Fusobacterium nucleatum, actinomycetes actinomyces, Actinomyces viscous, Pseudomonas fusiformis, Treponema pallidum, Intermediate bacillus and gingival bacillus were used as detection antigens, and the antibody titers against the periodontal disease-causing pathogen-specific IgY were detected. The results are shown in the following table

IgY detection antigen antibody titer

Porphyromonas gingivalis 1: 512

Fusobacterium nucleus 1: 512

Specific anti-periodontal pathogens Actinomycetes Actinomyces 1: 512

Sexual Chicken IgY Sticky Actinomyces 1: 512

Hold 1: 256

Tartar Treponema pallidum 1: 128

Intermediate Bacilli 1: 128

Gingival bacteria 1; 128

Porphyromonas gingivalis 1: 512

Fusobacterium nucleus 1: 512

Anti-periodontal pathogen-specific actinomycetes actinomycetes 1: 512 Sexual duck IgY sticky actinomyces 1: 512 Fusarium 1: 256 Treponema pallidum 1: 128 Intermediate bacillus 1: 128 Bacteroides gingivalis 1: 128 Porphyromonas gingivalis 1: 256 Fusobacterium nucleatum 1 : 256 anti-periodontal pathogenic actinomycetes actinomycetes 1: 256 specific goose IgY sticky actinomycetes 1: 256 Fusarium bacillus 1: 128 tartar Treponema pallidum 1: 64 intermediate bacillus 1: 64 gum Bacterium 1: 64 Porphyromonas gingivalis 1: 128 Clostridium nucleus 1: 128 Anti-periodontal pathogens Actinomycetes Actinomyces 1: 128 Specific turkey IgY Sticky actinomycetes 1: 128 Fu S. bacillus 1: 64. Treponema pallidum 1: 32. Intermediate bacillus 1: 32. G. gingivalis 1: 32. Porphyromonas gingivalis 1: 128 Clostridium nucleus 1: 128 Anti-periodontal pathogenic actinomycetes Symbiotic Actinomyces 1: 128 Specific Ostrich IgY Sticky Actinomyces 1: 128 m 1: 64 Treponema pallidum 1: 32 Intermediate Bacterium 1: 32 Gliobacillus 1: 32 It can be seen from the above test results that the method of the present invention is used to prepare antigens, the immune activation method is used to immunize hens, ducks, geese, laying turkeys, or laying ostriches, and then the methods of the present invention are used to prepare antibodies. Periodontal disease pathogen-specific chicken IgY, periodontal disease pathogen-specific duck IgY, periodontal disease pathogen-specific geese IgY, periodontal disease pathogen-specific turkey IgY, resistance to teeth There are five different IgY of ostrich-specific ostrich IgY, which are different from those of immunized birds. However, the specific IgY prepared has strong inhibitory effects on several pathogenic bacteria that cause periodontal disease. Although the titer of IgY antibodies produced by immune laying ostriches and laying turkeys is slightly lower from the test data, because the ostrich and fire eggs are particularly large, the IgY that can be extracted from each immune egg is higher than that of chickens. There are much more immune eggs, duck immune eggs, and goose immune eggs, which is also very desirable for industrial large-scale production. Experimental Example 4: The composite antigen prepared in Experimental Example 2 was used to immunize laying hens, laying hens, laying geese, laying turkeys, and laying ostriches. Then, the method of the present invention is used to prepare: (1) anti-periodontal pathogen-specific chicken IgY, (2) anti-periodontal pathogen-specific duck IgY, (3) anti-periodontal pathogen Specific goose IgY, (4) anti-periodontal pathogen-specific turkey IgY, (5) anti-periodontal pathogen-specific ostrich pupae. Then, the "ELISA method" (enzyme-linked immunosorbent assay) was used to test the bacterial species Gingivale, Intermediate, Bacillus fusiformis, Treponema pallidum, Actinomyces viscosus, Actinomycetes, and Fusobacterium nucleatum 2. Porphyromonas gingivalis is used as a detection antigen, and the antibody titers of these five kinds of periodontal disease-causing pathogen-specific IgY are detected. The results are shown in the following table

IgY detection antigen antibody titer

Gingival Bacteria 1: 512

Intermediate Bacterium 1: 512

Specific anti-periodontal pathogens entangled 1: 1 512

Sexual Chicken IgY Tartar Treponema pallidum 1: 512

Actinomyces viscosus 1: 256

Symbiotic Actinomycetes 1: 128

Fusobacterium nucleus 1: 128

Porphyromonas gingivalis 1: 128

Gingival Bacteria 1: 512

Intermediate Bacterium 1: 512

Specific anti-periodontal pathogenic bacteria Forsythia 1: 512

Sexual Duck IgY Tartar Treponema pallidum 1: 512 Actinomyces viscosus 1: 256

Symbiotic Actinomycetes 1: 128

Fusobacterium nucleus 1: 128

Porphyromonas gingivalis 1: 128

Gingival Bacteria 1: 256

Intermediate Bacilli 1: 256

Anti-periodontal pathogens Namba 1: 256

Specific Goose IgY Tartar Treponema pallidum 1: 256

Actinomyces viscosus 1: 128

Actinomycetes Actinomyces 1: 64

Fusobacterium nucleus 1: 64

Porphyromonas gingivalis 1: 64

Dental silver bacillus 1: 128

Intermediate Bacilli 1: 128

Anti-periodontal pathogens Expansion bribe 1 »1: 128

Specific Turkey IgY Tartar Treponema pallidum 1: 128

Actinomyces viscosus 1: 64

Actinomyces Symbiotic Actinomyces 1: 32

Fusobacterium nucleus 1; 32

Porphyromonas gingivalis 1:32

Gingival Bacteria 1: 128

Intermediate Bacilli 1: 128

Anti-periodontal pathogens ffi race cage 1: 128

Specific Ostrich IgY Tartar Treponema pallidum 1: 128

Actinomyces viscosus 1: 64

Actinomyces Symbiotic Actinomyces 1: 32

Fusobacterium nucleus 1: 32

Porphyromonas gingivalis 1:32 As shown in the table above, the antigen ratio is changed. The specific compound of various egg-laying poultry extracted after intensive immunization IgY antibody binding titer also changes accordingly, and the immune response is very good. Experimental Example 5: Patients with periodontal disease use the anti-periodontal disease-specific composite IgY of the present invention as a raw material to prepare a combined oral spray and short stick, and perform therapeutic observation.

(1) Twenty patients with gingivitis and 20 patients with periodontitis were randomly selected from patients with periodontal disease.

(2) Patients with gingivitis use oral spray daily after brushing their teeth, after mouthwash, after meals, at intermediate times, and before going to bed, spraying the spray on the gums and teeth 4-6 times a day, 4-6 times a day, on the teeth Patients with periodontitis (periodontal pocket 0-2mm) first apply oral spray, and then insert short sticks or IgY granules into the periodontal pocket. After each meal, clean one's mouth and one before bedtime. A total of 4 times.

(3) Observe the changes of gums, periodontal pockets and plaque.

(4) 7-15 days after administration, the symptoms of 20 patients with gingivitis basically disappeared. The gums changed from dark red to normal red, swelling and bleeding disappeared, and burning sensation disappeared.

In 15 patients with periodontitis, no pus was squeezed within 1-5 days after medication, the pain disappeared, the percussion turned negative, and the periodontal pockets were closed. The other 5 patients' symptoms were significantly alleviated, the plaque was significantly lightened, and no new plaques developed.

(5) The symptoms of gingivitis disappeared in 20 people for 15 consecutive days, periodontitis (periodontal pocket formation 0-2mm), 20 people, 18 people closed the periodontal bag, gingival swelling and bleeding disappeared, mobility improved, plaque disappeared or Significantly lightened, 1 person had slight swelling of the gingival papilla, and 1 person had mild plaque. Example 1: Preparation of 100 g of periodontal disease-specific IgY

(1) Selecting good breed ducks with high immune response ability:

Porphyromonas gingivalis was used to prepare antigens, and 1,000 female ducks were immunized, each time was injected with lral antigen, after the first injection, it was injected again every two weeks for a total of three times. At the 7th month after the first injection, the eggs produced by these ducks were labeled, and their IgY were extracted according to conventional methods. The titers of the IgY produced were detected by ELISA (enzyme-linked immunosorbent assay); 200 ducks with an IgY titer of 256 were produced, and then the eggs produced by this batch of ducks were used to hatch excellent duck breeds, which were allowed to grow up to 2-3 months as the preferred special female ducks with high immune response ability;

(2) Preparation of periodontal disease pathogen complex antigen

According to the culture characteristics and optimal culture conditions of Porphyromonas gingivalis, Clostridium nucleus, actinomycetes, Actinomyces viscosus, Pseudomonas fusiformis, Treponema pallidum, Intermediate bacillus, Gingival bacilli Select appropriate media respectively to multiply and expand the strains, and then harvest the culture for purification, counting of strains, and diluting and packing; refer to the occurrence rate of periodontal disease plaque with reference to the single periodontitis plaque after the culture, purification, and packing of the single antigen. And the proportion, mixed as follows: Porphyromonas gingivalis 20-5%

Fusobacterium nucleus 20-5%

Actinomyces actinomycetes 20-5%

Actinomyces viscous 15-10%

Forsythia bacillus 10-20%

Tartar Treponema pallidum 5-15%

Intermediate Bacteria 5-20%

Tooth root 5-20%

Then add the Freund's adjuvant at a ratio of 1: 1 to the mixed bacterial solution containing 2.0-3.5 billion bacteria counts / 0.5ml, and then put it into a high-speed homogenizer to homogenize at 10000-30000r / min to make it uniform. / 0 emulsion, to prepare periodontal disease pathogens complex antigen.

(3) Reinforce immunity on the preferred special female ducks:

Using the prepared periodontal pathogenic bacteria complex antigen, the combination of subcutaneous injection and wing vein injection was used to immunize the preferred special female ducks. The injection volume of each female duck reached 1 ml of antigen. On the 7th day and the 14th day, the injections were similarly performed once for strengthening. Twenty days after the first immunization, picking of immunized duck eggs began until the 10th month and termination and elimination of female ducks.

(4) Preparation of specific periodontal disease-resistant IgY:

First wash the immune eggs, sterilize and dry them with alcohol; break the immune eggs with an eggbeater, filter the egg whites with an egg yolk sieve, leave the egg yolks, and stir well; add 4-6 times distilled water and mix well; use 1. ON Adjust the pH of the HCL solution to 6.0-6.5, and stir well evenly. Let stand overnight at 4-6 ° C. Centrifuge this dilution at 2000r / min for 20 minutes. Take the supernatant with a cut-off molecular weight of 7-10 million. The ultrafiltration membrane was concentrated to 1/20; the concentrated solution was added with 0.1-0.3% sodium alginate to mix thoroughly, and then centrifuged overnight at 12000 rpm for 20 minutes; the supernatant was subjected to ultrafiltration; and then the membrane filtration technology was used for sterilization; The anti-periodontal disease-specific crude IgY extract was obtained; the ion-exchange column and the gel exchange column were used to construct the anti-periodontal disease-specific crude IgY extract to obtain pure IgY. Example 2: Preparation of an anti-periodontal oral spray

Prepare 100 liters of periodontal disease-specific IgY combination solution according to the following formula

Anti-periodontal disease-specific IgY

Sweet oil

Mannitol

Menthol 300g peppermint oil

Eucalyptus Leaf Oil 100g

Liquor 2000ml

Distilled water

Made into 100 liters

Preparation Process-

1. Take 80% of the distilled water, add mannitol and stir to dissolve. Then add IgY to dissolve and stir well. Adjust the pH to 6.5-7.5 with 1mol / L NaOH.

2. Add menthol, peppermint oil, eucalyptus oil, and ethanol to dissolve and add glycerin to mix well.

3. Stir again and slowly add 2 items to 1 item, and stir well.

4. Filter, add distilled water from the filter to the full amount, check the clarity, and fill the normal pressure spray after the pH titer is qualified. Each bottle is 20ml. Packed after passing the full inspection. Example 3: Preparation of Granules for Periodontal Bags

Formula:

Anti-periodontal Specific Compound IgY 100g

Hydroxypropyl methylcellulose (HPMC) 400g

Hydroxypropyl methylcellulose phosphate (HPMCP) 400g

Colloidal fine powder silicone 20g

Polyethylene glycol 4000 (PEG 4000) 80g

Acetone

1000g

Preparation Process:

1. Mix the specific composite IgY, HPMC, HPMCP colloidal Si0 ₂ and PHG4000 evenly and pre-mix twice through a 30 mesh sieve;

2. Add appropriate amount of acetone to make suitable soft materials;

3. Load it into a No. 9 syringe, extrude it, remove the solvent at room temperature, and then remove the solvent under vacuum;

4. Cut the above molding into thin strips so that the weight is between 1.5-2mg, and seal the package to obtain the periodontal disease-resistant composite IgY granules, which are used for periodontal pockets to remove plaque and anti-inflammatory treatment.

Example 4: Preparation of a specific periodontal disease-resistant composite IgY lozenge

formula: Anti-periodontal Specific IgY 3.8 g

Sorbitol 505.5 g

Maltodextrin 225 g

Aspartame 0.9 g

Menthol 1.5 g

Thin oil 0.98 ml

Eucalyptus Leaf Oil 0.33 ml

2% hydroxypropyl methylcellulose 50 ° /. Ethanol solution

Ethanol

Makes 1000 tablets

Preparation Process:

1. Sorbitol, maltodextrin and abbastame are sieved through 80 meshes, mixed according to the formula ratio, and sieved once through 60 meshes;

2. Using 2% hydroxypropyl methylcellulose 50% ethanol as a binder, add the above mixed powder to make suitable soft materials, granulate with 16 mesh nylon sieve, 60 # ventilate and dry, the moisture content of the particles is 3-5%. Granules, sieved through a 14-mesh sieve, sieve out an appropriate amount of fine powder, add IgY powder and mix well, and mix into the granules and stir;

3. Dissolve menthol, peppermint oil, eucalyptus oil and ethanol (approximately 5ml), spray into the above granules and seal for 4 hours, add magnesium stearate (1% by total), mix and press, each tablet weighs About 0.75g.

Claims

Rights request

1. An anti-periodontal disease-specific IgY, characterized in that the preparation method of the anti-periodontal disease-specific IgY includes the following steps: ''

Isolation and culture of the major pathogens causing periodontal disease, preparation of single or composite periodontal pathogen antigens; preparation of immune eggs; preparation of anti-periodontal disease-specific crude IgY extracts; purification of anti-periodontal disease-specific crude IgY extracts ; Provide a variety of anti-periodontal disease-specific IgY preparations.

2. The anti-periodontal disease-specific IgY according to claim 1, wherein the preparation method of the anti-periodontal disease-specific IgY comprises the following steps:

Based on the screening and classification of these pathogens, single or composite periodontal pathogen antigens are prepared respectively: according to the culture characteristics and optimal culture conditions of these pathogenic microorganisms, appropriate media are selected respectively for large-scale breeding and expansion of strains. Then the culture is harvested for purification, strain counting, dilution and packaging; the cultured pathogens are mixed individually or in an appropriate ratio, and then Freund's adjuvant is added separately in a 1: 1 ratio or other appropriate ratio, and placed in a high-speed homogenizer Homogenize at medium and high speed to form an oil-in-water solution, that is, make a single or composite periodontal pathogen antigen;

Preparation of immune eggs: Select laying egg SPF healthy hens with high immune response ability, and inject single or compound periodontal pathogen pathogen antigens at multiple points in the chest muscle or subcutaneously, and after the first injection, interval 7 Day, then the second injection with the same dose and method; the third injection with the same dose at an interval of 7 days after the second injection, and 20 days after the first injection, according to the titer of the complex IgY in serum and yolk Pick high titer immune eggs and mark the immune eggs with different antigens;

Preparation of crude anti-periodontal pathogen-specific composite IgY: Soak or wipe poultry eggs produced by hens immunized with different antigens separately, rinse them with sterile pure water and dry them, and place them in eggbeaters. Break the eggshell, filter the egg white with a yolk sieve, remove the egg yolk, add 5-8 times distilled water to dilute and mix, adjust the pH to 5.5-6.0 with Imol NaOH or HCL solution, and let stand at 4-6 ° C overnight. dilutions 1000- 3000r / min centrifugation for 20 minutes, the supernatant was concentrated 15-20 fold with D _a 5-10 million molecular weight cutoff ultrafiltration membrane, the concentrate was dried by freeze-drying, each single antigen IgY lyophilized powder Mix according to the ratio of 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10 to obtain anti-periodontal disease specific compound IgY crude; anti-periodontal Purification of crude pathogen-specific IgY: The crude anti-periodontal pathogen-specific IgY prepared by the above process was passed through an ion exchange column and a gel chromatography column, respectively, to obtain pure IgY.

3. The anti-periodontal disease-specific IgY according to claim 1 or 2, characterized in that the pathogens causing periodontal disease in the method for preparing periodontal disease-specific IgY include: Porphyromonas gingivalis, Clostridium nucleus, actinomycete actinomycetes, actinomyces viscosus, Pseudomonas fusiformis, Treponema pallidum, Intermediate bacterium, and gingival bacterium.

4. The periodontal disease-specific IgY according to claim 1 or 2, characterized in that the egg-producing poultry used for preparing immune eggs in the method for preparing periodontal disease-specific IgY includes hens and female ducks. , Mother goose, turkey, ostrich.

5. The periodontal disease-specific IgY according to claim 2, wherein the preparation of the composite periodontal pathogen antigen in the method for preparing periodontal disease-specific IgY is as follows: And dilute the prepared pathogens in 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10 and so on. The adjuvant was placed in a homogenizer at a speed of 10,000 to 30,000 r / min to homogenize it to become a uniform W / 0 emulsion, and a composite periodontal pathogen pathogen was prepared.

6. The anti-periodontal disease-specific IgY according to claim 5, characterized in that the components in the composite periodontal pathogen antigen are mixed in the following percentages:

Porphyromonas gingivalis 20-5% Fusobacterium nucleus 20-5%

Actinomyces actinomycetes 20-5%

Sexual Actinomyces 15-10%

Forsythia bacillus 10-20%

Tartar Treponema pallidum 5-15%

Intermediate Bacteria 5-20%

Gingival bacteria 5-20%

7. The periodontal disease-specific Y according to claim 1 or 2, characterized in that 0.02-80% of specific IgY is matched with 99.98-20% of medicinal excipients, which can be made into a clinically acceptable oral cavity. Mouth spray, mouthwash, lozenge, stick treatment or toothpaste.

8. An anti-periodontal disease-specific chicken IgY, characterized in that the preparation method of the anti-periodontal disease-specific chicken IgY includes the following steps:

Selecting good breed hens with high immune response ability: Porphyromonas gingivalis was used to make antigens, and 1,000 hens were immunized separately, each injection of 1 ml of antigen, after the first injection, re-injected every two weeks, a total of Three times, at the 7th month after the first injection, the eggs produced by these hens were labeled, and their IgY were separately extracted according to conventional methods; the effectiveness of the produced IgY was measured by ELISA (enzyme-linked immunosorbent assay) Select 200 hens with IgY titer of 256, and then use the eggs produced by this batch of hens to hatch excellent chicken breeds, and allow them to grow to 2-3 months as the preferred high immune response ability. Special hen

Preparation of Periodontal Pathogen Complex Antigen: According to Porphyromonas gingivalis, Clostridium nucleus, Actinomyces actinomyces, Actinomyces viscosus, Forsythia, Treponema pallidum, Intermediate bacteria, Gingivals The culture characteristics and optimal culture conditions of Bacillus spp., Select appropriate media respectively for large-scale breeding and expansion of strains, and then harvest the culture for purification, strain counting, and dilution and packing; the above cultures are purified and packed into a single antigen With reference to the probability and proportion of periodontal disease plaque, mix as follows:

Porphyromonas gingivalis 20-5%

Fusobacterium nucleus 20-5%

Actinomyces actinomycetes 20-5%

Actinomyces viscous 15-10%

Forsythia bacillus 10-20%

Tartar Treponema pallidum 5-15%

Intermediate Bacteria 5-20%

Gingival bacteria 5-20%

Then add the Freund's adjuvant at a ratio of 1: 1 to a mixed bacterial solution containing 2.0-3.5 billion bacteria counts / 0.5ml, and then put it into a high-speed homogenizer to homogenize at 10000-30000r / min to make it uniform. / 0 emulsion, to prepare periodontal disease pathogens complex antigen;

Preparation of immune eggs: using the prepared periodontal pathogenic bacteria complex antigen, using a combination of subcutaneous injection and wing vein injection, to individually immunize the preferred special hens, the injection amount of each hen reaches lml antigen, 7 days and 14 days after injection, the same injection once again for boosting, 20 days after the first immunization, open the platform to pick up the immune eggs until the 10th month to terminate and eliminate the hens;

Preparation of anti-periodontal disease-specific IgY: first wash the immune eggs, sterilize and dry them with alcohol; break the immune eggs with an eggbeater, filter the egg whites with an yolk sieve to remove the egg whites, leave the yolks, and stir them; add 4- 6 times distilled water, mix and hook; adjust the pH to 6.0 0 with 5.5N HCL solution, stir the hook evenly, stand at 4-6 ° C overnight, and centrifuge this dilution at 2000r / min for 20 minutes Take the supernatant and concentrate it with an ultrafiltration membrane with a molecular weight cut-off of 70,000 to 100,000, and concentrate it to 1/20; add 0.1-0.3% sodium alginate to the concentrate and mix thoroughly, centrifuge overnight at 12,000 rpm for 20 minutes; take the supernatant and ultrafiltration; then Use membrane filtration technology to sterilize; freeze-dry to obtain anti-periodontal specific IgY crude extract; The anti-periodontal disease-specific crude IgY extract was purified to obtain pure IgY.

9. A periodontal disease-specific duck IgY, characterized in that the preparation method of the periodontal disease-specific duck IgY includes the following steps:

Selecting good breed ducks with high immune response ability: Use Porphyromonas gingivalis to make antigens and immunize 1,000 female ducks each with 1 ml of antigen each time. After the first injection, reinject it every two weeks for a total of three times. In the seventh month after the first injection, the eggs produced by these ducks were labeled, and their IgY were extracted according to conventional methods. The titer of the produced IgY was measured by ELISA (enzyme-linked immunosorbent assay); Select 200 ducks with an IgY titer of 256, and then use the eggs produced by this batch of ducks to hatch excellent duck breeds, and wait for them to grow to 2-3 months as the preferred special female ducks with high immune response ability; Preparation of Periodontal Pathogen Complex Antigen: According to Porphyromonas gingivalis, Clostridium nucleus, Actinomyces actinomyces, Actinomyces viscosus, Forsythia, Treponema pallidum, Intermediate bacillus, Gingivals The culture characteristics and optimal culture conditions of Bacillus spp., Respectively, select appropriate mediums for large-scale breeding and expansion of strains, and then harvest the culture for purification, strain counting, and dilution and packing; divide the above cultures into purification and packing Single antigen refers to the probability and proportion of periodontal disease plaque, and is mixed as follows:

Porphyromonas gingivalis 20-5%

Fusobacterium nucleus 20-5%

Actinomyces actinomycetes 20-5%

Actinomyces viscous 15-10%

Forsythia bacillus 10-20%

Tartar Treponema pallidum 5-15%

Intermediate Bacteria 5-20%

Gingival bacteria 5-20%

Then one hundred million the number of bacteria containing 20-35 / 0.5ml mixed bacteria in a 1: 1 ratio Jiaru Fu's adjuvant, and then into a high-speed homogenizer to 10000-30000r / mi _n homogenized to a homogeneous W / 0 emulsion to prepare periodontal disease pathogenic complex antigen;

Preparation of immune eggs: using the prepared periodontal pathogenic bacteria composite antigen and subcutaneous injection of wing vein injection combined immunization method, the specific special female ducks are immunized, and the injection volume of each female duck reaches 1-2ml Antigens were boosted again on the 7th and 14th days after injection. 20 days after the first immunization, picking of immune duck eggs was started until the 10th month and the female ducks were eliminated.

Preparation of anti-periodontal disease specific IgY: First, wash the immune eggs, sterilize and dry them with alcohol; break the immune eggs with an eggbeater, filter the egg whites with an egg yolk sieve, leave the egg yolks, and stir well; add 4-6 Distilled water, mix well; adjust the pH to 6.0-6.5 with 1.0N H solution, stir well evenly, let stand overnight at 4-6 ° C, and centrifuge this dilution at 2000r / min for 20 minutes; Take the supernatant and concentrate it with an ultrafiltration membrane with a molecular weight cut-off of 70,000-100,000, and concentrate it to 1/20; add 0.1-0.3% sodium alginate to the concentrate and mix thoroughly, and overnight at 4 ° C, 12000rpn! Centrifuge for 20 minutes; ; Then use membrane filtration technology to sterilize; freeze-dry to obtain anti-periodontal disease-specific crude IgY extract; pass ion exchange column gel exchange column, anti-periodontal disease-specific crude IgY extract to obtain Pure IgY.

10. An anti-periodontal disease-specific goose IgY, which is characterized in that the preparation method of the anti-periodontal disease-specific goose IgY includes the following steps-selecting a good breed goose with high immune response ability: using gingival porphyrin alone The bacilli were used to make antigens, and 1,000 female geese were immunized. Each injection of 1 ml of antigen was given. After the first injection, the injections were repeated every two weeks for a total of three times. After the eggs produced by the goose are labeled, their IgY are separately extracted according to conventional methods; the produced IgY titers are detected by ELISA (enzyme-linked immunosorbent assay); 200 female geese with an IgY titer of 256 are selected, and then Use the eggs produced by these geese to hatch excellent goose breeds and wait for them to grow up to 2-3 months as the preferred special geese with high immune response ability;

Preparation of Periodontal Pathogen Complex Antigen: According to Porphyromonas gingivalis, Clostridium nucleus, Actinomyces actinomyces, Actinomyces viscosus, Forsythia, Treponema pallidum, Intermediate bacillus, Gingivals Cultivation characteristics and For the most suitable culture conditions, select the appropriate culture medium for large-scale breeding and expansion of the strains, and then harvest the culture for purification, strain counting, and dilution and packing; refer to the periodontal disease plaque with the single antigen purified and packed as above The probability and proportion of the occurrences are mixed as follows:

Porphyromonas gingivalis 20-5%

Fusobacterium nucleus 20-5%

Actinomyces actinomycetes 20-5%

. Actinomycetes 15-10%

Forsythia bacillus 10-20%

Tartar Treponema pallidum 5-15%

Intermediate Bacteria 5-20%

Gingival bacteria 5-20%

Then add the Freund's adjuvant at a ratio of 1: 1 to the mixed bacterial solution containing 2.0-3.5 billion bacteria counts / 0.5ml, and then put it into a high-speed homogenizer to homogenize at 10000-30000r / min to make it uniform. / 0 emulsion, to prepare periodontal disease pathogens complex antigen;

Immunize the preferred special goose separately: using the prepared periodontal pathogenic bacteria complex antigen, using a combination of subcutaneous injection and wing vein injection to immunize the preferred special goose, Each injection of female geese reached 2-10 ml of antigen, and the injections were performed on the 7th day and the 14th day after the injection for the same boosting. 20 days after the first immunization, the immune goose eggs were started to be picked up until the 10th month. Knock out geese

Preparation of anti-periodontal disease specific IgY: First, wash the immune eggs, sterilize and dry them with alcohol; break the immune eggs with an eggbeater, filter the egg whites with an egg yolk sieve, leave the egg yolks, and stir well; add 4- 6 Distilled water twice, mix well; use 1.0! ^} 1 (^ solution to adjust 11 to 6. () -6. 5, stir well evenly, let stand overnight at 4-6, centrifuge this dilution 2000r / min 20 minutes; Take the supernatant and concentrate it to 1/20 with a cut-off molecular weight of 70,000-100,000 ultrafiltration membrane; add 0.1-0.3% sodium alginate to the concentrate and mix thoroughly, centrifuge at 4 ° C overnight, and centrifuge at 12O0Orpm for 20 minutes; take the supernatant Ultrafiltration; followed by sterilization by membrane filtration technology; freeze-drying to obtain a periodontal disease-specific crude IgY extract; passing ion exchange columns and gel exchange columns to connect periodontal disease-specific crude IgY extracts To obtain pure IgY.

11. A periodontal disease-specific turkey IgY, which is characterized in that the preparation method of the periodontal disease-specific turkey IgY includes the following steps:

Selecting good breed egg-laying turkeys with high immune response ability: Use Porphyromonas gingivalis to make antigens, immunize 1,000 egg-laying turkeys each time, and inject 1 ml of antigen each time. After the first injection, every two weeks Inject once, three times in total, and in the 7th month after the first injection, the egg labels produced by these laying turkeys have been extracted separately according to conventional methods; ELISA method (Enzyme-linked immunosorbent assay) Test the prepared IgY titers; select 200 egg-laying turkeys with IgY titers of 256, and use the eggs from this batch of egg-laying turkeys to hatch excellent turkey species until they grow to 2 — 3 months, as a preferred special egg laying turkey with high immune response;

Preparation of Periodontal Pathogen Complex Antigen: According to Porphyromonas gingivalis, Clostridium nucleus, Actinomyces actinomyces, Actinomyces viscosus, Forsythia, Treponema pallidum, Intermediate bacillus, Gingivals The culture characteristics and optimal culture conditions of Bacillus spp., Select appropriate media respectively for large-scale breeding and expansion of strains, and then harvest the culture for purification, strain counting, and dilution and packing; the above cultures are purified and packed into a single antigen With reference to the probability and proportion of periodontal disease plaque, mix as follows-Porphyromonas gingivalis 20-5%

Fusobacterium nucleus 20-5%

Actinomyces actinomycetes 20-5%

Actinomyces viscous 15-10%

Forsythia bacillus 10-20%

Tartar Treponema pallidum 5-15%

Intermediate Bacteria 5-20% Gingival bacteria 5-20%

Then add the Freund's adjuvant at a ratio of 1: 1 to a mixed bacterial solution containing 2 to 3.5 billion bacterial counts / 0.5ml, and then put it into a high-speed homogenizer to mix at 10000-30000r / min to make it uniform. / O emulsion, to prepare periodontal disease pathogenic complex antigen;

Immunize the preferred special egg-laying turkeys separately: use the obtained periodontal disease pathogen complex antigen, and use subcutaneous injection and wing vein injection to immunize the preferred special egg-laying turkeys, The injection volume of each egg-producing turkey reached 3 15ml of antigen, and the injection was also performed on the 7th and 14th days after the injection to strengthen it. After 20 days of the first immunization, the immune fire eggs were picked up to the 10th month Stop and phase out laying turkeys;

Preparation of specific periodontal disease-resistant IgY: First, wash the immune eggs, sterilize and dry them with alcohol; break the immune eggs with an eggbeater, filter the egg whites with an egg yolk sieve, leave the egg yolks, and stir well; force 4 -6 times distilled water, mix well; adjust the pH to 6.0-6.5 with 1.0N HCL solution, stir well, stand still at 4-6 ° C overnight, and centrifuge this dilution at 2000r / min for 20 minutes Take the supernatant and condense it with a cut-off molecular weight of 70,000-100,000 ultrafiltration membrane to 1/20; add the concentrated solution with 0.1-0.3% sodium alginate and mix thoroughly, centrifuge at 4 ° C overnight, 12000rpm for 20 minutes; take the supernatant and ultrafiltration ; Then use membrane filtration technology to sterilize; freeze-dry to obtain anti-periodontal disease-specific crude IgY extracts; pass ion-exchange columns and gel-exchange columns to entangle anti-periodontal disease-specific crude IgY extracts, That is pure IgY.

12. An anti-periodontal disease-specific ostrich IgY, characterized in that the preparation method of the anti-periodontal disease-specific ostrich IgY includes the following steps:

Selecting good breeding egg-laying ostriches with high immune response ability: Use Porphyromonas dendrobium to make antigens, immunize 1,000 egg-laying ostriches each with 1 ml of antigen each time, and re-inject every two weeks after the first injection Once, three times in total, and in the 7th month after the first injection, the egg labels produced by these laying ostriches were recorded and their IgYs were extracted according to conventional methods. The ELISA method was used to detect The prepared IgY titer; 200 egg-laying ostriches with an IgY titer of 256 were selected, and then the eggs produced by this batch of egg-producing ostriches were used to hatch excellent ostrich species, which were allowed to grow up to 2-3 months , As a special egg-laying ostrich with preferred pupa immune response ability;

Preparation of periodontal disease pathogen complex antigen: According to Porphyromonas gingivalis, Clostridium nucleus, actinomycete actinomyces, sticky actinomycetes, Fusarium, Treponema pallidum, Intermediate bacillus, gums The culture characteristics and optimal culture conditions of Bacillus spp., Respectively, select the appropriate culture medium for large-scale breeding and expansion of strains, and then harvest the culture for purification, strain counting, and dilution and packing; the above cultures are purified and packed into a single antigen With reference to the probability and proportion of periodontal disease plaque, mix as follows:

Porphyromonas gingivalis 20-5%

Fusobacterium nucleus 20-5%

Actinomyces actinomycetes 20-5%

Actinomyces viscous 15-10%

Forsythia bacillus 10-20%

Tartar Treponema pallidum 5-15%

Intermediate Bacteria 5-20%

Gingival bacteria 5-20%

Immunize the preferred special egg-laying ostriches separately: using the prepared periodontal pathogenic bacteria complex antigen, using a combination of subcutaneous injection and wing vein injection to immunize the preferred special egg-laying ostriches, each Only ostrich ostrich injections reached 3-15ml of antigen per injection, and the injection was strengthened again on the 7th and 14th days after injection. After 20 days of the first immunization, the ostrich eggs were picked up until the 10th month and terminated. Eliminate laying ostriches;

Preparation of anti-periodontal disease specific IgY: First, wash the immune eggs, sterilize and dry them with alcohol; break the immune eggs with an eggbeater, filter the egg whites with an egg yolk sieve, leave the egg yolks, and stir well; add 4-6 0N Distilled water, mix well; use 1. 0N Adjust the pH of the HCL solution to 6.0-6.5, and stir well. Let stand overnight at 4-6 ° C. Centrifuge this dilution at 2000r / min for 20 minutes. Take the supernatant with a molecular weight cut-off of 7-10 million. The ultrafiltration membrane was concentrated to 1/20; the concentrated solution was added with 0.1-0.3% sodium alginate and mixed thoroughly, and centrifuged overnight at 12,000 rpm for 20 minutes; the supernatant was subjected to ultrafiltration; and then the membrane filtration technology was used for sterilization; Anti-periodontal disease-specific crude IgY extracts were obtained;

Anti-periodontal disease specific IgY crude extracts are modified to obtain pure IgY.

Eucalyptus camellia

13. A U preparation method for U-Ho-Lo anti-foliar periodontal disease oral spray, characterized in that the method is:

According to the following formula, adjust the alcohol and oil into 100% by volume of periodontal disease-specific IgY combination solution.

Heterosexual IgY 150 parts by weight

1500 parts by weight

200 parts by weight

300 parts by weight

100 parts by weight

2000 parts by volume

100000 parts by volume

Preparation Process:

Take a total of 80% of distilled water and add mannitol to stir and dissolve, then add IgY to dissolve and stir, and adjust the pH to 6.5-7.5 'with 1mol / L NaOH; menthol, add peppermint oil, eucalyptus oil, ethanol and stir to dissolve and add glycerin. Mix evenly; slowly add the latter item to the preceding item under stirring, and stir well; filter, add distilled water from the filter to the full amount, check the clarity, and pass the filling at normal pressure spray after the pH titer is qualified. In the bottle, each 20ml, packed after the full inspection.

14. A method for preparing granules for periodontal pockets, characterized in that the method is:

Formulation-100% by weight of periodontal disease-specific complex IgY

Hydroxypropyl methyl cellulose (HPMC) 400 parts by weight

Hydroxypropyl methylcellulose phosphobenzene: 400 parts by weight of formate (HPMCP)

20 parts by weight of colloidal fine powder silica gel

Polyethylene glycol 4000 (PEG 400O) 80 parts by weight

Propanone

1000 parts by weight

Preparation Process:

The specific complexes IgY, HPMC, HPMCP glue PHG4000 Si0 ₂ and mixed - with the premix through a 30 mesh screen twice; of acetone was added an appropriate amount of a suitable flexible material; Nine loaded syringe, extrusion molding, room temperature The solvent was removed under vacuum, and then the solvent was removed under vacuum. The molded article was cut into thin strips so that the weight was between 1.5 and 2 mg, and the package was sealed to obtain anti-periodontal composite IgY granules.

15. A method for preparing periodontal disease-specific composite IgY lozenges, characterized in that the method is:

Formula:

Periodontal disease specific complex IgY 3.8 parts by weight

Sorbitol 505.5 parts by weight

Maltodextrin 225 parts by weight

Aspartame 0.9 parts by weight

Menthol 1.5 Peppermint oil 0.98 vol.

Eucalyptus Leaf Oil 0.33 vol.

2% light propyl methyl cellulose 50% ethanol

Right amount

1000 tablets

Preparation Process:

Sorbitol, maltodextrin, and abbastame were sieved through an 80-mesh sieve, mixed uniformly according to the formula ratio, and sieved once through a 60-mesh sieve;

Using 2% hydroxypropyl methylcellulose and 50% ethanol as a binder, add the above mixed powder to make suitable soft materials, granulate on a 16 mesh nylon sieve, ventilate and dry at 60 ° C, and collect 3-5% granule moisture. After passing through a 14-mesh sieve, sieving an appropriate amount of fine powder, adding IgY powder and mixing, and mixing into the granules to mix well;

Menthol, peppermint oil, eucalyptus oil and ethanol (approximately 5 parts by volume) are dissolved, sprayed on the above granules and sealed for 4 hours, then add magnesium stearate (1% by total), mix and press, each tablet weigh About 0.75 parts by weight.

16. A method for preparing periodontal disease-resistant toothpaste, characterized in that the method is:

Add an appropriate amount of specific periodontal compound IgY to toothpaste and mix to obtain.