CN112826931A - Preparation method of anti-HPV and anti-gynecological inflammation composite ovovitellin gel - Google Patents
Preparation method of anti-HPV and anti-gynecological inflammation composite ovovitellin gel Download PDFInfo
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- CN112826931A CN112826931A CN202110014320.2A CN202110014320A CN112826931A CN 112826931 A CN112826931 A CN 112826931A CN 202110014320 A CN202110014320 A CN 202110014320A CN 112826931 A CN112826931 A CN 112826931A
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- 206010061218 Inflammation Diseases 0.000 title claims abstract description 47
- 230000004054 inflammatory process Effects 0.000 title claims abstract description 44
- 239000002131 composite material Substances 0.000 title claims abstract description 34
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 241000701806 Human papillomavirus Species 0.000 claims abstract description 32
- 241000700605 Viruses Species 0.000 claims abstract description 23
- 150000002632 lipids Chemical class 0.000 claims abstract description 13
- 239000002671 adjuvant Substances 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 13
- 239000000843 powder Substances 0.000 claims description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 8
- 241000287828 Gallus gallus Species 0.000 claims description 7
- 230000000840 anti-viral effect Effects 0.000 claims description 7
- 235000013330 chicken meat Nutrition 0.000 claims description 7
- 230000000694 effects Effects 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 7
- 235000013601 eggs Nutrition 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 239000002245 particle Substances 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 4
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 4
- 229920002125 Sokalan® Polymers 0.000 claims description 4
- 229960001631 carbomer Drugs 0.000 claims description 4
- 235000010445 lecithin Nutrition 0.000 claims description 4
- 229940067606 lecithin Drugs 0.000 claims description 4
- 239000000787 lecithin Substances 0.000 claims description 4
- 239000003755 preservative agent Substances 0.000 claims description 4
- 230000002335 preservative effect Effects 0.000 claims description 4
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims description 3
- 241000588724 Escherichia coli Species 0.000 claims description 3
- 241000341655 Human papillomavirus type 16 Species 0.000 claims description 3
- 241000191940 Staphylococcus Species 0.000 claims description 3
- 241000194017 Streptococcus Species 0.000 claims description 3
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 claims description 3
- 230000001804 emulsifying effect Effects 0.000 claims description 3
- 210000003527 eukaryotic cell Anatomy 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 210000001236 prokaryotic cell Anatomy 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 238000005086 pumping Methods 0.000 claims description 3
- 239000008213 purified water Substances 0.000 claims description 3
- 238000007790 scraping Methods 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000007599 discharging Methods 0.000 claims description 2
- 238000004108 freeze drying Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 241001631646 Papillomaviridae Species 0.000 abstract description 4
- 210000000270 basal cell Anatomy 0.000 abstract description 3
- 238000002560 therapeutic procedure Methods 0.000 abstract description 3
- 206010008342 Cervix carcinoma Diseases 0.000 description 3
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 3
- 201000010881 cervical cancer Diseases 0.000 description 3
- 208000009608 Papillomavirus Infections Diseases 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 241000722343 Human papillomavirus types Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010058846 Ovalbumin Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 210000003679 cervix uteri Anatomy 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000001571 immunoadjuvant effect Effects 0.000 description 1
- 239000000568 immunological adjuvant Substances 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 229940092253 ovalbumin Drugs 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 210000004994 reproductive system Anatomy 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/42—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum viral
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
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- A—HUMAN NECESSITIES
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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Abstract
The embodiment of the invention discloses a preparation method of an anti-HPV and anti-gynecological inflammation composite ovomyosin gel, which is characterized in that anti-HPV and anti-gynecological inflammation composite ovomyosin is compounded through anti-human papilloma virus ovomyosin to form an antibody cocktail therapy, and IgY antibody is carried and protected by a nano lipid carrier to directly reach basal cells, so that Human Papilloma Virus (HPV) and gynecological inflammation virus can be effectively neutralized.
Description
Technical Field
The embodiment of the invention relates to the technical field of medical treatment and health, and in particular relates to a preparation method of an anti-HPV and anti-gynecological inflammation composite ovoluteolin gel.
Background
The cervical cancer is a gynecological malignant tumor with the incidence rate of 2 nd next to that of breast cancer in China, and is the only cancer with definite etiology in the world at present, Human Papilloma Virus (HPV) infection is generally considered to be an important factor causing cervical canceration at present, but HPV infection is not the only factor causing cervical canceration, and besides HPV infection, chronic inflammation of the cervix caused by the reproductive system is also an independent risk factor inducing the cervical cancer. Therefore, elimination of female gynecological inflammation pathogenic bacteria and HPV virus is the most effective measure for effectively preventing cervical cancer. The embodiment of the invention provides a preparation method of an anti-HPV and anti-gynecological inflammation composite ovovitellin gel, which can effectively neutralize Human Papilloma Virus (HPV) and gynecological inflammation virus.
Disclosure of Invention
Therefore, the embodiment of the invention provides a preparation method of the anti-HPV and anti-gynecological inflammation compound ovovitellin gel, which can effectively neutralize Human Papilloma Virus (HPV) and gynecological inflammation virus.
In order to achieve the above object, the embodiments of the present invention provide the following technical solutions: a preparation method of an anti-HPV and anti-gynecological inflammation composite ovovitellin gel, which comprises the following steps:
1) screening typical human papilloma virus and typical gynecological inflammation virus;
2) injecting the obtained high-activity high-purity human papilloma virus and gynecological inflammation virus of different types into SPF chickens with adjuvants respectively, and exciting to generate antiviral ovomyosin aiming at different types of viruses, and obtaining eggs containing different antiviral ovomyosin;
3) crushing, separating, purifying and freeze-drying the obtained eggs, and mixing to obtain high-purity anti-HPV and anti-gynecological inflammation composite ovomyosin freeze-dried powder;
4) through high-speed rotation, the anti-HPV and anti-gynecological inflammation composite ovomyosin freeze-dried powder is wrapped in a nano lipid carrier;
5) sequentially adding the composite ovoglobulin freeze-dried powder, carbomer, preservative and auxiliary materials wrapped in the nano lipid carrier into an emulsifying machine, and stirring by a scraping frame type stirrer and a paddle type stirrer until the mixture is uniformly stirred and has no bubbles to obtain the gel.
Further, screening typical human papilloma virus specifically comprises:
the virus early genes E6 and E7 are expressed in eukaryotic cells or prokaryotic cells, self-assembled into virus-like particles and purified to obtain the purified virus-like particles, namely the human papilloma virus.
Further, the human papillomavirus types include HPV6, HPV11, HPV16, HPV18, HPV31, HPV33, HPV45, HPV52, HPV58, HPV65 and HPV 81.
Further, the gynecological inflammation virus comprises staphylococcus, streptococcus, escherichia coli and candida.
Further, the adjuvant includes an R71 adjuvant, a freund adjuvant, an aluminum salt adjuvant and the like.
Further, the nano lipid carrier comprises lecithin.
Further, the auxiliary materials comprise glycerol and purified water.
Further, the method further comprises:
in the step 5), a vacuum pump is used for discharging bubbles in the raw materials in the stirring process, and the liquid level position in the tank body is noticed during operation, so that the vacuum pump is prevented from pumping the raw materials out.
The embodiment of the invention has the following advantages:
according to the preparation method of the anti-HPV and anti-gynecological inflammation composite ovomyosin gel provided by the embodiment of the invention, anti-HPV and anti-gynecological inflammation composite ovomyosin is compounded through anti-human papilloma virus ovomyosin to form an antibody cocktail therapy, and IgY antibody is carried and protected by a nano lipid carrier to directly reach basal cells, so that Human Papilloma Virus (HPV) and gynecological inflammation virus can be effectively neutralized.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
FIG. 1 shows the detection result of the HPV antigen activity of the anti-HPV and anti-gynecological inflammation compound ovomyosin freeze-dried powder composition provided in example 1 of the invention;
FIG. 2 shows the result of the detection of the anti-HPV and anti-gynecological inflammation complex ovomyosin gel combined with HPV antigen activity provided in example 1 of the present invention.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The embodiment 1 of the invention provides a preparation method of an anti-HPV and anti-gynecological inflammation composite ovovitellin gel, which comprises the following steps:
1) typical human papilloma virus and typical gynecological inflammation virus are screened.
The virus early genes E6 and E7 are expressed in eukaryotic cells or prokaryotic cells, self-assembled into virus-like particles and purified to obtain the purified virus-like particles, namely the human papilloma virus.
In this example, the types of human papillomaviruses include HPV6, HPV11, HPV16, HPV18, HPV31, HPV33, HPV45, HPV52, HPV58, HPV65, and HPV 81. The gynecological inflammation virus includes Staphylococcus, Streptococcus, Escherichia coli, and Candida.
2) Injecting the obtained high-activity high-purity human papilloma virus and gynecological inflammation virus of different types into SPF chicken together with adjuvants respectively, and exciting to generate antiviral ovomyosin against different types of viruses to obtain eggs containing different antiviral ovomyosin.
In this embodiment, the composite immunoadjuvant includes R71 adjuvant, freund's adjuvant, aluminum salt adjuvant, and the like.
Each virus, type, was injected independently, and each chicken received only one virus injection, resulting in only one antiviral ovalbumin.
3) The obtained eggs are crushed, separated, purified, freeze-dried and mixed to obtain the high-purity anti-HPV and anti-gynecological inflammation composite ovomyosin freeze-dried powder.
Each chicken only produces single antibody corresponding to the injection type, and the single ovoglobulin is respectively purified, lyophilized and mixed to obtain the composite ovoglobulin. When the virus is injected into the chicken, the adjuvant is added, so that the antibody produced in the chicken is the most and the activity is the highest.
4) The anti-HPV and anti-gynecological inflammation composite ovomyosin freeze-dried powder is wrapped in the nano lipid carrier through high-speed rotation.
In this example, lecithin was used as the nano lipid carrier. The antibody is wrapped in a nano lipid carrier so as to facilitate membrane penetration and better absorption.
5) Sequentially adding the composite ovoglobulin freeze-dried powder, carbomer, preservative and auxiliary materials wrapped in the nano lipid carrier into an emulsifying machine, and stirring by a scraping frame type stirrer and a paddle type stirrer until the mixture is uniformly stirred and has no bubbles to obtain the gel.
In this embodiment, the auxiliary materials include glycerin, purified water, and the like. In the step 5), a vacuum pump is used for exhausting bubbles in the raw materials in the stirring process, and the liquid level position in the tank body is noticed during operation, so that the vacuum pump is prevented from pumping the raw materials out.
In the embodiment, the dosage percentage of each component is as follows: 0.1-3% of anti-human papilloma virus ovovitellin (IgY), 0.1-3% of anti-gynecological inflammation ovovitellin (IgY), 0.5-5% of carbomer and the balance of preservative and auxiliary materials.
Examples of the experiments
The neutralization effect of the anti-HPV and anti-gynecological inflammation composite ovoglobulin lyophilized powder composition and the prepared anti-HPV and anti-gynecological inflammation composite ovoglobulin gel on HPV virus is detected by an Elisa method, and as shown in figures 1 and 2, the test results show that the composite ovoglobulin composition and gel in the embodiment can effectively neutralize human papilloma virus HPV.
According to the preparation method of the anti-HPV and anti-gynecological inflammation composite ovomyosin gel provided by the embodiment of the invention, anti-HPV and anti-gynecological inflammation composite ovomyosin is compounded through anti-human papilloma virus ovomyosin to form an antibody cocktail therapy, and IgY antibody is carried and protected by a nano lipid carrier to directly reach basal cells, so that Human Papilloma Virus (HPV) and gynecological inflammation virus can be effectively neutralized.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (8)
1. A preparation method of an anti-HPV and anti-gynecological inflammation composite ovovitellin gel is characterized by comprising the following steps:
1) screening typical human papilloma virus and typical gynecological inflammation virus;
2) injecting the obtained high-activity high-purity human papilloma virus and gynecological inflammation virus of different types into SPF chickens with adjuvants respectively, and exciting to generate antiviral ovomyosin aiming at different types of viruses, and obtaining eggs containing different antiviral ovomyosin;
3) crushing, separating, purifying and freeze-drying the obtained eggs, and mixing to obtain high-purity anti-HPV and anti-gynecological inflammation composite ovomyosin freeze-dried powder;
4) through high-speed rotation, the anti-HPV and anti-gynecological inflammation composite ovomyosin freeze-dried powder is wrapped in a nano lipid carrier;
5) sequentially adding the composite ovoglobulin freeze-dried powder, carbomer, preservative and auxiliary materials wrapped in the nano lipid carrier into an emulsifying machine, and stirring by a scraping frame type stirrer and a paddle type stirrer until the mixture is uniformly stirred and has no bubbles to obtain the gel.
2. The method for preparing the anti-HPV and anti-gynecological inflammation composite ovomyosin gel according to claim 1, characterized in that, the typical human papilloma virus is screened, and the method specifically comprises the following steps:
the virus early genes E6 and E7 are expressed in eukaryotic cells or prokaryotic cells, self-assembled into virus-like particles and purified to obtain the purified virus-like particles, namely the human papilloma virus.
3. The method for preparing the anti-HPV and anti-gynecological-inflammation composite ovovitellin gel according to claim 1, wherein the types of human papillomavirus comprise HPV6, HPV11, HPV16, HPV18, HPV31, HPV33, HPV45, HPV52, HPV58, HPV65 and HPV 81.
4. The method for preparing the anti-HPV and anti-gynecological inflammation composite ovovitellin gel according to claim 1, wherein the gynecological inflammation viruses comprise staphylococcus, streptococcus, escherichia coli and candida.
5. The method for preparing the anti-HPV and anti-gynecological inflammation composite ovovitellin gel according to claim 1, wherein the adjuvant comprises R71 adjuvant, Freund's adjuvant, aluminum salt adjuvant, etc.
6. The method for preparing the anti-HPV and anti-gynecological-inflammation composite lecithin gel according to claim 1, wherein the nano lipid carrier comprises lecithin.
7. The method for preparing the anti-HPV and anti-gynecological-inflammation composite ovovitellin gel according to claim 1, wherein the auxiliary materials comprise glycerol and purified water.
8. The preparation method of the anti-HPV and anti-gynecological inflammation composite ovomyosin gel according to claim 1, characterized in that the method further comprises:
in the step 5), a vacuum pump is used for discharging bubbles in the raw materials in the stirring process, and the liquid level position in the tank body is noticed during operation, so that the vacuum pump is prevented from pumping the raw materials out.
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| CN116554280A (en) * | 2023-05-18 | 2023-08-08 | 上海博满生物科技有限公司 | 18-valent HPV composite yolk neutralizing antibody for preventing and treating cervical HPV infection, and preparation method and application thereof |
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