CN113730573A - Antiviral composition and application thereof - Google Patents

Antiviral composition and application thereof Download PDF

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CN113730573A
CN113730573A CN202010461844.1A CN202010461844A CN113730573A CN 113730573 A CN113730573 A CN 113730573A CN 202010461844 A CN202010461844 A CN 202010461844A CN 113730573 A CN113730573 A CN 113730573A
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composition
igy
acid
yolk antibody
virus
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夏铮
黄玉吉
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Jiaxing Baoran Biotechnology Co ltd
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Jiaxing Baoran Biotechnology Co ltd
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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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/351—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom not condensed with another ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Virology (AREA)
  • Public Health (AREA)
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  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
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Abstract

The invention discloses an antiviral composition and application thereof. The composition comprises biological active components such as yolk antibody (IgY) and bird's nest acid (Neu5Ac), and has the effects of inhibiting hemagglutinin, regulating cellular immunity, inhibiting cytokine storm and the like through certain compatibility, so that the composition can prevent and treat related viral diseases such as influenza, viral pneumonia and the like.

Description

Antiviral composition and application thereof
Technical Field
The invention belongs to the health field of foods, health care products and medicines, and particularly relates to a composition containing bioactive components for preventing and treating viral diseases.
Background
Influenza (called influenza for short) is acute respiratory tract infection caused by influenza virus and is also a disease with strong infectivity and high transmission speed. It is transmitted primarily by airborne droplets, human-to-human contact, or contact with contaminated items. Typical clinical symptoms are: acute high fever, general pain, marked weakness and mild respiratory symptoms. Generally, the autumn and winter season is the high-incidence period of the disease, and the complications and death phenomena caused by the disease are very serious.
Influenza is caused by influenza virus (fig. 1), and is classified into three types, i.e., a (a), B (B), and C (C), and a D (D) type, which has been discovered in recent years. The type of a particular influenza virus is determined by antigenic variation, namely two glycoproteins on the viral surface: hemagglutinin (HA) and Neuraminidase (NA). Both glycoproteins are targets for anti-influenza virus drug development. Among them, oseltamivir, which is widely used clinically at present, is an anti-influenza drug with neuraminidase as an action target. Hemagglutinin (HA), which is accidentally discovered by Hirst in the study of influenza virus, is a spike-shaped glycoprotein on the envelope surface of influenza virus and plays an important role in the processes of releasing virus from host cells into cytoplasm after invading host cell nucleus, and the like. At present, the anti-influenza virus candidate drug Fladase (influenza enzyme) with hemagglutinin as the action target has been in the phase IIb of clinical test in the United states. The action mechanism is to block the combination of the fragment HA1 after the enzyme digestion of the virus surface hemagglutinin and a host cell membrane surface sialic acid receptor.
The novel coronavirus pneumonia (abbreviated as 'new coronavirus pneumonia') is pneumonia caused by infection of the novel coronavirus (abbreviated as 'new coronavirus'). On day 11/2 of 2020, the world health organization always wins the report and names pneumonia infected by new coronavirus as "COVID-19" (Corona Virus Disease 2019). Meanwhile, the International Committee for Classification of viruses states that the new Coronavirus was named "SARS-CoV-2" (Severe acid Respiratory Syndrome Coronavir 2).
The new coronavirus is a novel coronavirus. The virus surface expresses envelope protein (E), protuberant protein (S), membrane protein (M) and Hemagglutinin Esterase (HE). The S protein mediates the mutual fusion of the new coronavirus envelope and the host cell membrane, and is a key target for the current drug development. The hemagglutinin esterase (fig. 1) is used as an adsorption protein to help the virus to be adsorbed to the surface of the host cell at an early stage, determines the range of the host cell, and can also cause erythrocyte agglutination, generate adsorption on erythrocytes and the like. Therefore, the hemagglutinin esterase also has very important significance for developing anti-new coronavirus medicines.
On the other hand, infection of a host with a virus causes a series of natural immune responses, and the host promotes the production of cytokines such as INF, IL-1. beta., IL-18, etc. through pathways such as RLR, TLR, NLR, etc., to protect against the virus. However, highly pathogenic viruses, such as influenza virus H5N1, new corona virus, etc., can utilize the loopholes of the host's natural immune system to cause cytokine imbalance in the body, which results in lung tissue damage and systemic disease.
According to the first clinical data report of new coronavirus infection cases published in the "Lancet" section, after the new coronavirus attacks the human body, the immune system of the human body is disturbed, and various cell factors in body fluid, such as TNF-alpha, IL-1, IL-6, IL-12, IFN-alpha, IFN-beta, IFN-gamma and the like, are rapidly and massively produced. These abundantly expressed cytokines are important causes of acute respiratory distress syndrome and multiple organ failure.
Researchers in Chongqing Sanxia center hospitals analyzed the cell immunity and cytokine status of 123 patients with new coronary pneumonia, and predicted the relationship between the cell immunity level and cytokine and the disease condition of the patients. The research shows that the number of CD4+, CD8+ and B cells and NK cells in the new coronary pneumonia patients is obviously reduced. Among them, the numbers of CD4+ and CD8+ were significantly different between the patients with mild or severe cases, while the numbers of B cells and NK cells were not significantly different. Meanwhile, the levels of cytokines such as IL-6, IL-17, TNF, IFN, IL-4 and the like in the body of a patient with new coronary pneumonia are obviously increased. Wherein, IL-6 is obviously increased in the process of the disease change from light to heavy, and IL-17, TNF, IFN and IL-4 have no significant difference.
Therefore, in addition to blocking the interaction between the virus and the host and inhibiting the virus replication, the scientific treatment scheme for diseases such as influenza or new coronary pneumonia also needs to be capable of participating in the natural immune response of the body and inhibiting excessive inflammatory response, so as to avoid immune imbalance and tissue damage caused by the virus.
Therefore, there is an urgent need to develop drugs, health products and foods for inhibiting coronavirus and enhancing immunity.
Disclosure of Invention
The invention aims to provide a composition containing a bioactive ingredient for preventing and treating viral diseases.
Another object of the present invention is to provide a process for the preparation of the above composition.
It is a further object of the present invention to provide the use of the above composition for the preparation of a medicament for the prevention and treatment of viral infectious diseases.
Aiming at the above purpose, the invention provides the following technical scheme:
in one aspect, the present invention provides a composition for preventing and treating viral diseases, comprising a bioactive ingredient, which contains a yolk antibody (IgY) and a bird's nest acid (Neu5Ac) as active ingredients.
Wherein, the content of the yolk antibody (IgY) and the cubilose acid (Neu5Ac) in each gram of the composition is 1 mg-1 g of the yolk antibody (IgY); 1 mg-1 g of cubilose acid (Neu5 Ac); preferably 10 mg-1 g of yolk antibody (IgY); 10 mg-1 g of cubilose acid (Neu5 Ac).
Preferably, the weight ratio of the yolk antibody (IgY) to the cubilose acid (Neu5Ac) is 1: 0.01-100.
More preferably, the weight ratio of the yolk antibody (IgY) to the cubilose acid (Neu5Ac) is 1:100, 1:10, 1: 1.
The product type of the composition is any one of food, health care products and medicines.
The composition is taken orally, and comprises chewing or swallowing.
The composition has a daily oral dosage of more than 1 g.
Preferably, its daily oral dose is 1.5 grams.
The composition is in a dosage form selected from any one of tablets, powder, pills, granules, capsules or oral liquid.
In another aspect, the present invention provides a method for preparing a composition for preventing and treating viral diseases, comprising a bioactive ingredient, the method comprising the steps of:
(1) preparation of yolk antibody (IgY): selecting healthy egg-laying hens of a proper age, and immunizing the egg-laying hens with a specific antigen to obtain eggs containing specific IgY; sterilizing the obtained immune eggs, separating egg white and yolk to obtain yolk liquid containing specific IgY, and purifying to obtain yolk antibody;
(2) preparation of cubilose acid (Neu5 Ac): extracting natural nidus Collocaliae with cold water, or fermenting with specific engineering bacteria to obtain fermentation liquid, centrifuging, purifying, and spraying or lyophilizing to obtain nidus Collocaliae acid;
(3) mixing the yolk antibody (IgY) obtained in the step and the cubilose acid (Neu5Ac), and adding auxiliary materials to obtain the composition.
Wherein, the yolk antibody (IgY) and the cubilose acid (Neu5Ac) can also be obtained from the market; the auxiliary materials can be various conventional auxiliary materials required by preparing different dosage forms: diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, and the like.
In a further aspect, the present invention provides a use of the composition for the preparation of a medicament for preventing or treating a viral infectious disease.
Preferably, the viral infectious disease is selected from the group consisting of diseases caused by infection with coronavirus, influenza virus, parainfluenza virus, adenovirus, rhinovirus, respiratory syncytial virus, coxsackie virus or echovirus.
The invention has the advantages that:
(1) the yolk antibody and the cubilose acid are scientifically mixed for the first time to obtain the composition for preventing or treating the viral infection diseases;
(2) the composition has the effects of blocking virus infection of host cells and inhibiting virus replication;
(3) the composition also has the effects of regulating the cellular immunity of organisms, inhibiting cytokine storm and the like.
Drawings
FIG. 1 is a schematic structural diagram of an influenza virus.
FIG. 2 coronavirus surface-expressed proteins: envelope protein E, protuberant protein S, membrane protein M and hemagglutinin esterase HE.
Detailed Description
The embodiments of the present invention will be described in detail below with reference to the accompanying drawings so that the objects, features and advantages of the invention can be more clearly understood. It should be understood that the embodiments shown in the drawings are not intended to limit the scope of the present invention, but are merely intended to illustrate the spirit of the technical solution of the present invention.
In the following description, for the purposes of illustrating various disclosed embodiments, certain specific details are set forth in order to provide a thorough understanding of the various disclosed embodiments. One skilled in the relevant art will recognize, however, that the embodiments may be practiced without one or more of the specific details. In other instances, well-known devices, structures and techniques associated with this application may not be shown or described in detail to avoid unnecessarily obscuring the description of the embodiments.
Throughout the specification and claims, the word "comprise" and variations thereof, such as "comprises" and "comprising," are to be understood as an open, inclusive meaning, i.e., as being interpreted to mean "including, but not limited to," unless the context requires otherwise.
Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should be noted that the term "or" is generally employed in its sense including "and/or" unless the context clearly dictates otherwise.
Example 1 preparation of antiviral combinations
(1) Preparation of yolk antibody (IgY): selecting healthy egg-laying hens of a proper age, and immunizing the egg-laying hens with a specific antigen to obtain eggs containing specific IgY; sterilizing the obtained immune eggs, separating egg white and yolk to obtain yolk liquid containing specific IgY, and purifying to obtain yolk antibody;
(2) preparation of cubilose acid (Neu5 Ac): extracting natural nidus Collocaliae with cold water, or fermenting with specific engineering bacteria to obtain fermentation liquid, centrifuging, purifying, and spraying or lyophilizing to obtain nidus Collocaliae acid;
the mixing ratio of the yolk antibody (IgY) and the bird's nest acid can be adjusted according to the application, and the ratio of the yolk antibody (IgY) and the bird's nest acid is 1: 0.01-100, preferably 1:100, 1:10 and 1: 1.
Example 2 determination of binding force of Oriental acid and Hemagglutinin Esterase (HE) glycoprotein
The binding force between the cubilose acid and HE is measured by a bio-layer interference method (BLI). The HE protein is biotinylated and then fixed on a Sterpavidin (SA) probe, the association and dissociation conditions of the HE protein and the bird's nest acid are determined by using bird's nest acid with different concentrations, arbidol is used as a positive control group, and finally, the KD value of the association of the HE protein and the bird's nest acid is calculated to be 5.2 mu mol and the KD value of the arbidol is calculated to be 4.5 mu mol. Thus, the antiviral composition can bind to the HE protein and inhibit coronaviruses.
Example 3 Effect of antiviral composition on cytokine levels in vitro
In vitro experiments, compared with supernatants of cells of different groups, the virus-infected cells have obviously increased levels of cytokines such as IL-6, IL-17, TNF, IFN, IL-4 and the like, and the composition has obviously reduced levels of cytokines such as IL-6, IL-17, TNF, IFN, IL-4 and the like.
Example 4 control of influenza in mice by antiviral compositions
Example pharmacodynamic study of compositions against avian influenza Virus H7N7 in vitro
CPE method determination of half the toxic concentration of the virus on MDCK cells (TCID50)
MDCK cells (canine kidney epithelial cells, provided by infectious disease focus laboratory of department of health of the first hospital affiliated to the medical college of Zhejiang university) were seeded in 96-well plates, each well100 μ L of 2X 10 cells4Culturing at 37 deg.C for 24 hr, growing into monolayer, diluting avian influenza virus H7N7 strain (provided by infectious disease emphasis laboratory of department of health of the first hospital affiliated to Zhejiang university medical college) with 10 times of culture solution, and sequentially diluting by 10 times-3To 10-12A total of 10 concentrations were added to the wells, 8 wells per concentration, and incubated at 37 ℃. Cytopathic effect (CPE) was observed daily with an inverted microscope. The degree of cytopathic effect (CPE) was recorded. Calculation of half the viral infectious dose (TCID) by Reed-Muench method50)。
Swelling, net pulling, shrinkage and aggregation after cell infection with virus, TCID of avian influenza virus H7N750Is 10-4.19/0.1 ml.
Assay of test substances against influenza virus
MDCK cells were seeded in 96-well plates at 100. mu.L/well and contained 2X 10 cells4Per well, 37 ℃, 5% CO2After 24h of culture, the cells grew into a monolayer. Discarding the supernatant, mixing the drug solutions with equal volumes of 100TCID50Influenza viruses are mixed in a tube, and after 6 hours of action, the mixed solution is inoculated on a cell plate. A normal cell control group, a positive drug control group (Tamiflu, available from Roche pharmaceutical Co., Ltd.) and a virus control were set. Adsorbing for 2 hr, discarding supernatant, adding cell maintenance solution, and culturing at 37 deg.C in 5% CO2 incubator. Cytopathic results were recorded and cellular activity was measured by MTT staining. The experiment was repeated 2 times. Statistical software SPSS13.0 was used to perform Probit regression analysis on the data and calculate the half effective concentration (IC) of the drug50)。
The half inhibitory concentration (IC50) of the tested drug is respectively determined to be 410 mu g/mL and 447 mu g/mL, which indicates that the tested drug has the functions of resisting virus and protecting cells.
Examples preferred embodiments of the present invention have been described in detail above, but it should be understood that aspects of the embodiments can be modified, if necessary, to employ aspects, features and concepts of the various patents, applications and publications to provide yet further embodiments.
These and other changes can be made to the embodiments in light of the above detailed description. In general, in the claims, the terms used should not be construed to be limited to the specific embodiments disclosed in the specification and the claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled.

Claims (10)

1. A composition for preventing and treating viral diseases comprising a bioactive ingredient, wherein the composition comprises a yolk antibody (IgY) and a bird's nest acid (Neu5Ac) as active ingredients.
2. The composition of claim 1, wherein the yolk antibody (IgY) and the bird's nest acid (Neu5Ac) respectively comprise 1mg to 999mg of the yolk antibody (IgY); 1 mg-999 mg of cubilose acid (Neu5 Ac).
3. The composition according to claim 2, wherein the weight ratio of the yolk antibody (IgY) to the bird's nest acid (Neu5Ac) is 1: 0.01-100, preferably 1:100, 1:10, 1: 1.
4. The composition according to any one of claims 1 to 4, wherein the product type of the composition is any one selected from the group consisting of food, health product and pharmaceutical product.
5. The composition of any one of claims 1 to 5, wherein the composition is administered orally, including either by chewing or swallowing.
6. Composition according to claim 6, wherein the daily oral dose of the composition is above 1g, preferably 1.5 g.
7. The composition according to any one of claims 1 to 8, wherein the composition is in a dosage form selected from any one of tablets, powders, pills, granules, capsules and oral liquids.
8. The composition of claim 1, wherein the composition is prepared by a process comprising the steps of:
(1) preparation of yolk antibody (IgY): selecting healthy egg-laying hens of a proper age, and immunizing the egg-laying hens with a specific antigen to obtain eggs containing specific IgY; sterilizing the obtained immune eggs, separating egg white and yolk to obtain yolk liquid containing specific IgY, and purifying to obtain yolk antibody;
(2) preparation of cubilose acid (Neu5 Ac): extracting natural nidus Collocaliae with cold water, or fermenting with specific engineering bacteria to obtain fermentation liquid, centrifuging, purifying, and spraying or lyophilizing to obtain nidus Collocaliae acid;
(3) mixing the yolk antibody (IgY) obtained in the step and the cubilose acid (Neu5Ac), and adding auxiliary materials to obtain the composition.
9. Use of the composition according to any one of claims 1 to 9 in the preparation of a medicament for the prevention or treatment of a viral infectious disease.
10. The use according to claim 11, wherein the viral infectious disease is selected from the group consisting of diseases caused by infection with coronavirus, influenza virus, parainfluenza virus, adenovirus, rhinovirus, respiratory syncytial virus, coxsackie virus, echovirus, norovirus, enterovirus type 71, and the like.
CN202010461844.1A 2020-05-27 2020-05-27 Antiviral composition and application thereof Pending CN113730573A (en)

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