CN109602904A - A kind of immune globulin IgY plural gel agent for gynaecological imflammation - Google Patents
A kind of immune globulin IgY plural gel agent for gynaecological imflammation Download PDFInfo
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- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
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Abstract
The invention belongs to fields of biomedicine, and in particular to a kind of plural gel agent of gynaecological imflammation.It include composite yolk antibody and nano silver in the gelling agent.Wherein, composite yolk antibody is obtained by Gardnerella and Candida albicans as antigen direct immunization laying hen.Yolk antibody of the invention can be used for preparing the drug of gynaecological imflammation caused by treatment vagina infection, and vagina medicinal inputs high activity antibody to gynaecological imflammation patient.In conjunction with nano silver, the gynaecological imflammation plural gel agent of Yolk antibody of the invention and preparation is significant for the healing of gynecological disease caused by vagina infection.
Description
Technical field
The invention belongs to fields of biomedicine, and in particular to a kind of immunoglobulin plural gel agent of gynaecological imflammation.
Background technique
Bacterial vaginosis BV (bacterial vaginosis, BV) can increase the risk that women suffers from pelvic inflammatory disease, increase
Add the neurological susceptibility to sexually transmitted disease and leads to the consequences such as Disadvantage pregnancy.Gardnerella vaginalis (GV) is the weight of bacterial vaginosis BV
Want one of pathogenic bacteria.Gardnerella vaginitis is one kind also known as haemophilus property vaginitis of bacterial vaginosis BV, is by adding
Moral receives a kind of vagina mucosa inflammation caused by bacillus, and passability, which is handed over, to be infected.
Gardner bacillus is the tiny bacillus of Gram-negative, is often in club-shaped, sometimes in Filamentous and Multiple Shape, common the two poles of the earth
Dyeing.It is 0.4--0.6 microns wide, long 1--2 microns of size, inactivity, atrichia, without gemma.Many bacterial strains have pod membrane.In people
It must be accorded to new blood ability growth and breeding when work culture, therefore have the title of " bloodthirsty ".But its biological nature and haemophilus are not
Identical to the greatest extent, clinically caused vaginitis is nonspecific vaginitis.
Infection caused by Gardner bacillus is most lighter, and the property be more common in enlivens women.Acute stage leucorrhea increasing, have fish raw meat or
The stink of ammonia, vulva is moist uncomfortable, is often accompanied by vagina burning heat sensation, intercourse pain and pudendal pruritus.Inspection is shown in that some patientss vulva is red
Swollen, vagina mucosa is congested, is in bois de rose, and mild redness, secretion is in homogenieity more, thin, canescence, sometimes for milk yellow or
Class green, bad smell.Vagina pH is often 5--5.5.Sometimes leukorrhea amount is few, only thin layer, as film sample is covered on congested yin
On road wall.Small number of patients vaginal wall has erythema or petechiae, and pregnant patient can cause miscarriage or puerperal endometritis.Infect severe one
It may also lead to septicemia, urinary tract infection, perinephric abscess and cystitis etc..Using metronidazole or Moxifloxacin standard care, though face
Bed symptom disappears, but easy to recur after drug withdrawal.
It there is no the drug that can be treated Gardnerella vaginitis and prevent recurrence at present.Meanwhile vaginitis is more sometimes
Caused by kind pathogen mixed infection.
Therefore a kind of compound formulation of gynaecological imflammation is developed, ingredient includes the Yolk antibody of Gardnerella and pharmaceutically may be used
The carrier of receiving.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of medicines for being used to prepare gynecological disease caused by treatment vagina infection
The composition of object, formula is safe, simple, and the drug of preparation can reduce the recurrence rate of vaginitis, to Gardner easy to recur
Bacterium vagina infection cure rate is high.
To achieve the above object, the technical solution of the present invention is as follows:
A kind of composition for the drug being used to prepare gynecological disease caused by treatment vagina infection, which is characterized in that described group
The active constituent for closing object is composite yolk antibody；The composition also includes nano silver and glycerol；
The composite yolk antibody is obtained by Gardnerella, Candida albicans as antigen immunization laying hen.
Yolk antibody (Immunoglobulin of yolk, IgY) is by inoculation laying hen, by the egg of its production
Corresponding antibody is extracted in Huang, and can be used for the prevention and treatment of corresponding disease.
Main component in yolk is protein and fat.Most of protein is all lipoprotein, is present in yolk protein
In, not soluble in water, only livitin (α, β, γ) is water-soluble, and IgY is γ livitin.Therefore point of IgY
From purifying firstly the need of the lipid being effectively removed in yolk, IgY is separated from water-solubility protein.
Had been established many more efficient economic methods, these methods mostly with PEG, dextran sulfate, natural gum,
As the methods of glycan or ethanol precipitation preliminary purification protein are pitched in mosanom, angle.
The method of industrial large-scale production IgY has: supercritical carbon dioxide gas extraction process, carragheen method, ammonium sulfate
Salting out method.
Preferably, the composite yolk antibody, nano silver, glycerol proportion be 20-100 parts by weight: 0.2-1 parts by weight:
300-500 parts by volume.
Further, the composite yolk antibody, nano silver, glycerol proportion be 40-70 parts by weight: 0.2-0.3 parts by weight:
300-500 parts by volume.
The preparation method of above-mentioned composite yolk antibody the following steps are included:
1) Gardnerella and Candida albicans are cultivated, is crushed after mixing, is homogenized acquisition antigen；
2) the antigen immunization laying hen for obtaining step 1) takes the egg of the laying hen after being immunized, and obtains yolk；
3) composite yolk antibody of the Gardnerella and Candida albicans in yolk is purified.
Preferably, the mass ratio of Gardnerella described in step 1) and Candida albicans is 1:1.
Such as without special specified, the unit of weight of the parts by weight in the traditional chinese medicine composition of the invention formula and the volume list of parts by volume
Position is matching relationship.That is: when the unit of weight of parts by weight is Kg, the volume unit of parts by volume is L；When the weight list of parts by weight
When position is g, the volume unit of parts by volume is mL.
Composition of the invention can also be prepared any as needed for can directly give in the form of a solution when treating
Pharmaceutically acceptable dosage form.The pharmaceutically acceptable dosage form includes composition of the invention and optional one or more pharmacy
Upper acceptable carrier, diluent or excipient and step suitable during the preparation process addition.Term used in the present invention
" pharmaceutically acceptable " refers to such compound, raw material, composition and/or dosage form, the model that they judge in rational medicine
In enclosing, be suitable for contacting with patient tissue and without excessive toxicity, irritation, allergy or with reasonable interests/Hazard ratio phase
Symmetrical other problems and complication, and effective for given application.
Pharmaceutical preparation of the invention is suitable for local administration, can be made into ointment, cream, suspension, lotion, powder, molten
Liquor, paste, gelling agent, spray, aerosol, oil formulation or transdermal patch.
Preferably, the drug of the treatment gynecological disease of composition of the invention preparation further includes other active components.
Preferably, the gynecological disease of the drug therapy of the treatment gynecological disease of composition of the invention preparation is mycotic yin
Vaginitis caused by road inflammation, trichomonas vaginitis, bacterial vaginitis, gonococcal vaginitis, vagina mixed infection.
Preferably, the gynecological disease is interior negative itch, cervical erosion, peculiar smell of smelling as of rotten fish, leucorrhea increasing caused by vagina infection.
The second object of the present invention is to provide a kind of gel for treating gynecological disease caused by vagina infection, can reduce
The recurrence rate of vaginitis is high to Gardnerella vagina infection cure rate easy to recur.
A kind of gel for treating gynecological disease caused by vagina infection, the formula of the gel are as follows: composite yolk antibody 40-
70 parts by weight, 1000ppm nano silver aqueous dispersion 200-300 parts by volume, glycerol 300-500 parts by volume, water 1500-2000
Parts by volume.
Further, preferably 50 parts by weight of composite yolk antibody, 200 parts by volume of 1000ppm nano silver aqueous dispersion,
400 parts by volume of glycerol, 1500 parts by volume of water；PH value is 5.5 ± 0.1.
The composite yolk antibody is immunized as antigen after being mixed by Gardnerella, Candida albicans by weight 1:1 and produces
Laying hen obtains.
The preparation method of above-mentioned composite yolk antibody the following steps are included:
1) Gardnerella and Candida albicans are cultivated, is crushed after mixing, is homogenized acquisition antigen；
2) the antigen immunization laying hen for obtaining step 1) takes the egg of the laying hen after being immunized, and obtains yolk；
3) composite yolk antibody of the Gardnerella and Candida albicans in yolk is purified.
The beneficial effects of the present invention are: the pharmaceutical composition of gynecological disease caused by treatment vagina infection provided by the invention
Object:
1) recurrence rate for reducing vaginitis is high to Gardnerella vagina infection cure rate easy to recur；
2) ingredient safety, antibiotic-free do not generate drug resistance；
3) directly outer to be used for infection site, therapeutic effect is good.
Specific embodiment
It detailed description of a preferred embodiment of the present invention will be given below.The reality of actual conditions is not specified in preferred embodiment
Proved recipe method, usually according to normal condition, illustrated embodiment are but not to be to preferably be illustrated to the contents of the present invention
The contents of the present invention are only limitted to illustrated embodiment.So those skilled in the art are according to foregoing invention content to embodiment party
Case carries out nonessential modifications and adaptations, still falls within protection scope of the present invention.
The specificity composite IgY antibody of the anti-gynecological inflammation pathogen of embodiment 1
1, prepared by antigen
Candida albicans, Gardnerella are absorbed from vaginitis patient's body, and uses conventional method culture.
Obtained Candida albicans, Gardnerella are mixed in the ratio of 1:1 and crushed with conventional method, is placed into
It is sufficiently smashed to pieces and is stirred evenly with 8000-12000rpm in " tissue mashing refiner ".Then by this mixed liquor with 1-10:1-10's
Freund adjuvant is added in ratio (usually 1:1), is placed in high speed homogenizer and is homogenized with 30000rpm high speed, forms water-in-oil emulsion,
Obtain complex antigen.
2, Antibody preparation
1) it is immunized:
1. the selected bird inlay with high immune response ability is made using immune activation method using step
The complex antigen obtained carries out injecting immune.It is spaced 7 days after first time injects, then for the second time with same dosage, method injection, the
It is spaced after biphasic injection 7 days and is injected the 3rd time with same dose again, pick high immune egg after injecting for the first time since the 20th day.
2) preparation of IgY crude product:
Respectively by produced high immune egg 0.5% benzalkonium bromide solution or 0.1%KMnO4 solution or other similar disinfections
Liquid, which impregnates immune egg 15-30 minutes, to be sterilized, and sterile distilled water is rinsed, dried, and is placed in eggbeater and is smashed eggshell, uses yolk
Sieving is added 4-8 times of distilled water dilution except egg white leaves yolk and stirs evenly, with 1mol/l NaOH liquid or 1mol/l HCL liquid
PH to 5.5-6.0 is adjusted, is stood overnight at 4-6 DEG C, dilution 8,000-12 000r/min high speed centrifugation 20 minutes, takes supernatant
Liquid is concentrated by ultrafiltration with ultrafilter.
Using the U.S. Pall Ultrafine Filtration Company manufacture except virus filtration system bacterial disease
Malicious filtering device thoroughly filters out various bacterial viruses, it is ensured that prepared IgY never contains any virus and bacterium.
First of bacterium filtering device is to remove bacteriums such as detection of Salmonella (Salmonella) with 0.22 μm of film sterilizing filter；
Second mycoplasma filtering device is to remove mycoplasma filter with 0.1 μm of film to remove mycoplasma；Third road virus filtering device is
A variety of viruses of the virus filter removing including avian influenza virus, enterovirus are removed with Ultipor VFTM DV50.
3) bioactivity:
It is above obtained using enzyme-linked immunization (ELISA) detection respectively using Candida albicans, Gardnerella as antigen
Anti- gynecological inflammation pathogen specific IgY semifinished product antibody combination potency.
Testing result is shown in Table 1:
1 bioactivity result of table
Testing result shows that the antibody combination potency of obtained anti-gynecological inflammation pathogen specific IgY is higher, completely
The requirement of medical treatment is reached.
4) prepared by IgY sterling:
Obtained anti-gynecological inflammation pathogen specific IgY crude product is crossed respectively ion exchange column and gel exchange column with
And affinity column is purified to get the pure IgY of anti-gynecological inflammation pathogen specific；
Obtained anti-gynecological inflammation pathogen specific IgY application freeze drier is dry, it is scorching to obtain anti-gynaecology
Disease pathogen specific IgY dry powder.
Obtained anti-gynecological inflammation pathogen specific IgY dry powder is taken, crushing of milling, processing is added in micronizer
At size between 1-100nm, granularity be more than >=15000 mesh ultramicron, corresponding nanosizing anti-gynaecological imflammation disease is made
Substance specific IgY dry powder.
Using SDS-PAGE electrophoresis assays, to the anti-gynecological inflammation pathogen specific IgY produced by process above
Sterling is detected, and measuring wherein IgY content is 98% or more.
Embodiment 2 treats the composition 1 of gynaecological imflammation and the composite yolk antibody gel of preparation
Composite yolk antibody 50g, 1000ppm the nano silver aqueous dispersion 200ml that embodiment 1 obtains, glycerol
400ml, water 1500ml；It is 5.5 ± 0.1 that pH value is adjusted after mixing, and homogenate obtains gel.
Embodiment 3 treats the composition 2 of gynaecological imflammation and the composite yolk antibody gel of preparation
Composite yolk antibody 80g, 1000ppm the nano silver aqueous dispersion 300ml that embodiment 1 obtains, glycerol
500ml, water 1500ml；It is 5.5 ± 0.1 that pH value is adjusted after mixing, and homogenate obtains gel.
Embodiment 4 treats the composition 3 of gynaecological imflammation and the composite yolk antibody gel of preparation
Composite yolk antibody 30g, 1000ppm the nano silver aqueous dispersion 200ml that embodiment 1 obtains, glycerol
300ml, water 1500ml；It is 5.5 ± 0.1 that pH value is adjusted after mixing, and homogenate obtains gel.
5 validity check of embodiment
1. clinical test
1. subjects: selection vaginitis patient 68, the age 20-58 years old.
2. testing process
Subjects are divided into experimental group and control group, use composite yolk antibody gel (embodiment 2) of the invention respectively
(35) and metronidazole suppository (33).
Collection of specimens: in the non-menstrual period sampling of patient, vagina innerlich anwenden is avoided before sampling.
Treatment cycle: experimental group uses gel of the invention, using 1 month, except after menstrual period and menstrual period 3 beyond the highest heavens, continuously
Using this product, daily 1, every 2.5g.It is checked after deactivating January, June.Control group uses external application metronidazole by operation instruction
Suppository is checked after deactivating January, June.
2. result:
1. composite yolk antibody gel test group clinical effectiveness is shown, turn out cloudy case load 30 after deactivating January, number of not turning out cloudy
5, negative conversion rate 85.71%；External application metronidazole suppository control group is turned out cloudy case load 30, is not turned out cloudy number 3, negative conversion rate
90.9%.
2. after deactivating June, composite yolk antibody gel test group feminine gender case history 15, external application metronidazole suppository control group yin
Venereal disease goes through 8.
As a result, composite yolk antibody of the invention and composite yolk antibody gel turn out cloudy for vaginitis patient's sun have it is important
Effect, and control recurrence rate after healing far below metronidazole suppository control group.It is easy to recur to solve vaginitis to a certain extent
Problem.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although referring to compared with
Good embodiment describes the invention in detail, those skilled in the art should understand that, it can be to skill of the invention
Art scheme is modified or replaced equivalently, and without departing from the objective and range of technical solution of the present invention, should all be covered at this
In the scope of the claims of invention.
Claims (10)
1. a kind of composition for the drug for being used to prepare gynecological disease caused by treatment vagina infection, which is characterized in that the combination
The active constituent of object is composite yolk antibody；The composition also includes nano silver and glycerol；
The composite yolk antibody is obtained by Gardnerella, Candida albicans as antigen immunization laying hen.
It is the composite yolk antibody, nano silver, sweet 2. composition according to claim 1-3, characteristic are
The proportion of oil is 20-100 parts by weight: 0.2-1 parts by weight: 300-500 parts by volume.
3. composition according to claim 1, characteristic are, the preparation method of the composite yolk antibody includes following
Step:
1) Gardnerella and Candida albicans are cultivated, is crushed after mixing, is homogenized acquisition antigen；
2) the antigen immunization laying hen for obtaining step 1) takes the egg of the laying hen after being immunized, and obtains yolk；
3) composite yolk antibody of the Gardnerella and Candida albicans in yolk is purified.
4. composition according to claim 3, which is characterized in that Gardnerella described in step 1) and Candida albicans
Mass ratio is 1:1.
5. the described in any item compositions of claim 1-4 answering in the drug that gynecological disease caused by vagina infection is treated in preparation
With, which is characterized in that the drug of the treatment gynecological disease can be any pharmaceutically acceptable dosage form.
6. the described in any item compositions of claim 1-4 answering in the drug that gynecological disease caused by vagina infection is treated in preparation
With, which is characterized in that the drug of the preparation treatment gynecological disease further includes other active components.
7. the described in any item compositions of claim 1-4 answering in the drug that gynecological disease caused by vagina infection is treated in preparation
With, which is characterized in that the gynecological disease is colpomycosis, trichomonas vaginitis, bacterial vaginitis, gonococcal vagina
Vaginitis caused by scorching, vagina mixed infection.
8. the described in any item compositions of claim 1-4 answering in the drug that gynecological disease caused by vagina infection is treated in preparation
With, which is characterized in that the gynecological disease is interior negative itch, cervical erosion, peculiar smell of smelling as of rotten fish, leucorrhea increasing caused by vagina infection.
9. a kind of gel for treating gynecological disease caused by vagina infection, which is characterized in that the formula of the gel is as follows: compound ovum
Yellow antibody 40-70 parts by weight, 1000ppm nano silver aqueous dispersion 200-300 parts by volume, glycerol 300-500 parts by volume, water
1500-2000 parts by volume；
The composite yolk antibody is used as antigen immunization laying hen after being mixed by Gardnerella, Candida albicans by weight 1:1
It obtains.
10. gel according to claim 9, which is characterized in that the formula of the gel is as follows: 50 weight of composite yolk antibody
Measure part, 200 parts by volume of 1000ppm nano silver aqueous dispersion, 400 parts by volume of glycerol, 1500 parts by volume of water；PH value is 5.5
±0.1。
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| CN110227156A (en) * | 2019-06-12 | 2019-09-13 | 华懿思科(成都)生物科技有限公司 | A kind of elimination gynaecological imflammation pathogenic bacteria and the lgY targeting preparation of HPV viruse and preparation method thereof |
| CN112358545A (en) * | 2020-11-10 | 2021-02-12 | 中国农业大学 | Preparation method and application of pigeon trichomonas and candida albicans bigeminal egg yolk antibody powder |
| CN117244057A (en) * | 2023-09-20 | 2023-12-19 | 江苏润洁生物科技有限公司 | Preparation method of clinical egg yolk antibody product for gynecological inflammation |
| CN117244057B (en) * | 2023-09-20 | 2024-05-14 | 江苏润洁生物科技有限公司 | Preparation method of clinical egg yolk antibody product for gynecological inflammation |
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