CN114903914B

CN114903914B - Application of gold nano material in inhibiting coronavirus

Info

Publication number: CN114903914B
Application number: CN202110183901.9A
Authority: CN
Inventors: 李洋; 张国芳
Original assignee: Shenzhen Institute of Advanced Technology of CAS
Current assignee: Shenzhen Institute of Advanced Technology of CAS
Priority date: 2021-02-10
Filing date: 2021-02-10
Publication date: 2023-03-28
Anticipated expiration: 2041-02-10
Also published as: WO2022170845A1; CN114903914A

Abstract

The invention relates to the application of gold nano-material in inhibiting coronavirus, and specifically discloses the application of gold nano-material in preparing a medicament for treating or preventing diseases caused by coronavirus or a material for adhering or inhibiting coronavirus; the coronavirus is selected from one of HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV2 or MERS-CoV; the disease caused by coronavirus is a disease caused by coronavirus infection. The material disclosed by the invention is simple to prepare, high in biological safety and excellent in effect.

Description

Application of gold nano material in inhibiting coronavirus

Technical Field

The invention belongs to the technical field of antivirus, and particularly relates to an application of a gold nano material in inhibiting coronavirus.

Background

Coronaviruses are a large family of viruses known to cause the common cold and more severe diseases such as Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). 2019 the novel coronavirus (SASR-CoV-2, causing novel coronavirus pneumonia COVID-19) is the 7 th known type at present, the first 6 are HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV (causing severe acute respiratory syndrome) and MERS-CoV (causing middle east respiratory syndrome), respectively, and no effective medicine exists at present.

At the present stage, nano-drugs and related preparations which can obviously inhibit the infection of coronavirus cells and have convenient sources and low cost are lacked. Moreover, few drugs can simultaneously achieve the therapeutic and prophylactic effects.

Disclosure of Invention

In order to overcome the defects in the prior art, the invention provides the application of the gold nano material in preparing the medicines for treating or preventing the diseases caused by the coronavirus in one aspect.

In the technical scheme of the invention, in the above application, the medicament further comprises a pharmaceutically acceptable carrier or excipient.

In the technical scheme of the invention, in the above application, the medicament further comprises administering a therapeutically effective amount of at least one other therapeutic agent or a composition thereof, wherein the other therapeutic agent or the composition thereof is selected from corticosteroids, anti-inflammatory signal transduction modulators, beta 2-adrenoceptor agonist bronchodilators, anticholinergics, mucolytics, hypertonic saline and other medicaments for treating coronavirus infection; or mixtures thereof.

In a second aspect, the present invention provides a pharmaceutical composition for treating or preventing a disease caused by coronavirus, wherein the pharmaceutical composition comprises a gold nanomaterial as an active ingredient.

In a technical scheme of the present invention, in the pharmaceutical composition, the pharmaceutical composition further includes a pharmaceutically acceptable carrier or excipient.

In the technical scheme of the invention, in the pharmaceutical composition, a second active ingredient is further included, and the second active ingredient is selected from corticosteroids, anti-inflammatory signal transduction modulators, beta 2-adrenoceptor agonist bronchodilators, anticholinergics, mucolytics, hypertonic saline and other drugs for treating coronavirus infection; or mixtures thereof.

In the technical scheme of the invention, in the pharmaceutical composition, the preparation form of the pharmaceutical composition is selected from oral preparations, injection preparations, mucosal administration preparations, inhalants and external preparations.

In a third aspect, the invention provides a method of treating or preventing a coronavirus infection, comprising administering to a subject a therapeutically effective amount of a gold nanomaterial.

In the technical scheme of the invention, in the above method, the gold nanomaterial with a therapeutically effective amount is administered in a preparation form, and the preparation further comprises a pharmaceutically acceptable carrier or excipient.

In one embodiment of the present invention, the method further comprises the step of administering to the subject a therapeutically effective amount of a second active ingredient selected from the group consisting of corticosteroids, anti-inflammatory signal transduction modulators, β 2-adrenoceptor agonist bronchodilators, anticholinergics, mucolytics, hypertonic saline, and other drugs used to treat infections with viruses of the family coronaviridae; or mixtures thereof.

The fourth aspect of the invention provides the use of gold nano-materials in the preparation of preparations for inhibiting coronavirus from reducing the combination of SARS-CoV-2 virus Spike protein and the receptor ACE2 of host cells thereof; preferably, the gold nanomaterial inhibits its binding to the ACE2 receptor of its host cell by competitively binding to the coronavirus Spike protein.

The fifth aspect of the invention provides the application of the gold nano material in preparing a material for inhibiting the infection of coronavirus or reducing and killing coronavirus.

In a sixth aspect, the invention provides a material for inhibiting coronavirus infection, the material comprising a gold nanomaterial and a matrix.

In the technical scheme of the invention, in the materials, the materials are used for preparing protective articles, and the protective articles are preferably masks, protective clothing, protective face masks and protective hats.

In the technical scheme of the invention, in the material, the material is a paint.

In the technical scheme of the invention, in the material, the material is a disinfectant or a personal care product.

In a technical solution of the present invention, in the above material, the material is a packaging material.

In the technical scheme of the invention, in the material, the material is a filter material.

According to a seventh aspect of the present invention, there is provided an article comprising a gold nanomaterial;

the article is selected from a protective article, a coating, a disinfectant, a personal care article, a packaging material, or a filtration device;

preferably, the protective articles are masks, protective clothing, protective face masks and protective hats;

preferably, the coating is a suspension containing gold nano-materials, and the coating can endow the surface of the attached object with a coating layer of the gold nano-materials;

preferably, the disinfectant is an environmental disinfectant, a no-clean hand sanitizer, a hand sanitizer and a detergent; more preferably, the disinfectant is used for surface disinfection of packaging materials, medical instruments, oral appliances, cosmetic appliances;

preferably, the packaging material is provided with a coating layer containing gold nano-materials, or the packaging material is a composite material containing the gold nano-materials; more preferably, the packaging material is a packaging material for pharmaceuticals or foodstuffs, especially for cold chain transportation;

preferably, the filtering device is a device for air filtration, a device for water filtration, a device for mask filtration; more preferably, the filtering device is an air filtering net, and the surface of the air filtering net is provided with a gold nano material; more preferably, the filtering device is air filter cotton, and gold nano-materials are attached to the fibers of the air filter cotton.

In the technical scheme of the invention, the coronavirus is selected from one of HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV2 or MERS-CoV.

In the technical scheme of the invention, the coronavirus is preferably SARS-CoV or SARS-CoV-2.

In the technical scheme of the invention, the disease caused by coronavirus is the disease caused by coronavirus infection.

In conclusion, compared with the prior art, the invention achieves the following beneficial effects:

1. the gold nano material can inhibit cell infection of coronavirus on one hand, and can specifically act on an infection target of the coronavirus on the other hand, so that the infection efficiency of the virus is reduced, the number of the coronavirus is reduced, and the gold nano material can be used as a therapeutic drug for the coronavirus.

2. Because the nanometer material of the invention inhibits the infection of SARS-CoV-2 virus to host cells, the specific adsorption and fixation of the virus are realized. Therefore, the coating can be used for preparing medicines, can be applied to melt-blown cloth, a refrigeration house coating and an outer packaging coating or spray of a mask, can adsorb new coronavirus, and plays a role in preventing new coronary pneumonia.

3. The gold nano material can be stored for a long time at room temperature to 4 ℃ and is convenient to store. The gold nano material has high stability, can be mixed and compounded with any material to prepare a composite material, and is not influenced by factors such as temperature, pH and the like.

4. The invention adopts inorganic nano materials, has unlimited material sources, is suitable for large-scale mass production, and has low cost and small using amount.

Drawings

Fig. 1 is a result of the detection of the physicochemical properties of the gold nanomaterials, wherein the left is a Transmission Electron Microscope (TEM) picture, and the right is a particle size distribution diagram.

FIG. 2 is the inhibitory effect of the gold nanomaterial of example 1 on the infection of ACE2/293T cells by pseudovirus SC2-P.

FIG. 3 is the effect of gold nanomaterials of example 1 on the inhibition of Euvirus SARS-CoV-2 infection into Vero-E6 cells.

Detailed Description

In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, shall fall within the scope of protection of the present invention.

Is widely applied in the fields of nano medicine and the like. At present, the effect of the nano-materials on inhibiting the infection of coronavirus, especially the cell infection of SARS virus SARS-CoV and new coronavirus SARS-CoV-2 is not reported. The invention proves that the gold nano material has high biological safety and low toxicity, and can be used in the fields of medicines, disinfectant, foods and materials.

Term(s) for

In the present invention, gold has the same meaning as Au.

In the present invention, the terms "nanomaterial" and "nanoparticle" have similar meanings and all refer to a material having one dimension in space that is nanoscale, such as nanoparticles having a particle size of 1-300nm or 1-100 nm. For example, the particle diameter is 1nm, 5nm, 10nm, 15nm, 20nm, 30nm, 40nm, 50nm, 60nm, 70nm, 80nm, or 90nm.

The ACE2 protein and angiotensin converting enzyme 2 have the same meaning, and research shows that the key of the novel coronavirus infection human body cell lies in the combination of the Spike protein of the coronavirus and the ACE2 protein on the surface of the human body cell and the entry into a host cell.

The S protein, spike protein or Spike glycoprotein represent the same meaning. Spike glycoproteins are glycoproteins protruding from the surface of the viral envelope that allow the virus to bind to and enter the cell.

"pharmaceutically acceptable carrier or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that has been approved by the food and drug administration as acceptable for use in humans or livestock.

"pharmaceutical composition" refers to a formulation of a compound of the present invention and a vehicle generally accepted in the art for delivering biologically active compounds to a mammal (e.g., a human). Such vehicles include all pharmaceutically acceptable excipients therefor.

An "effective amount" or "therapeutically effective amount" refers to an amount of a compound according to the present invention that, when administered to a patient in need thereof, is sufficient to effect treatment of a disease state, condition or disorder for which gold nanomaterials have utility. Such an amount will be sufficient to elicit the biological or medical response of the tissue system or patient sought by the researcher or clinician. The amount of a compound according to the invention which constitutes a therapeutically effective amount will vary depending on factors such as: the compound and its biological activity, the composition for administration, the time of administration, the route of administration, the rate of excretion of the compound, the duration of the treatment, the type of disease state or condition being treated and its severity, the drugs used in combination or in concert with the compounds of the invention, and the age, weight, general health, sex, and diet of the patient. Such therapeutically effective amounts can be routinely determined by those of ordinary skill in the art based on their own knowledge, the prior art, and the present disclosure.

The term "treating" as used herein, unless otherwise indicated, refers to reversing, alleviating, inhibiting the progression of, or preventing the disorder or condition to which this term applies, or one or more symptoms of such disorder or condition. The term "treatment" as used herein refers to the act of treating, as "treating" is as described immediately above. In some embodiments, the term "treating" is intended to mean administering gold nanomaterials according to the invention or compositions thereof to reduce or eliminate symptoms of coronavirus infection and/or reduce viral load in a patient.

"prevention" (or prophylaxis) refers to any treatment of a disease or condition that results in the failure of the clinical symptoms of the disease or condition to progress. The term "preventing" also includes administering a therapeutically effective amount of a compound or composition according to the invention prior to exposure of the individual to the virus (e.g., pre-exposure prevention) to prevent the onset of symptoms of the disease and/or to prevent the virus from reaching detectable levels in the blood.

The term "subject" or "patient" refers to an animal, such as a mammal (including a human), who has been or will be the object of treatment, observation or experiment. The methods described herein can be used for human therapy and/or veterinary applications. In some embodiments, the subject is a mammal (or patient). In some embodiments, the subject (or patient) is a human, a livestock animal (e.g., dogs and cats), a farm animal (e.g., cows, horses, sheep, goats, and pigs), and/or a laboratory animal (e.g., mice, rats, hamsters, guinea pigs, rabbits, cats, dogs, and monkeys). In some embodiments, the subject (or patient) is a human. "a human (or patient) in need thereof" refers to a human who may have or is suspected of having a disease or condition that would benefit from some treatment; for example, treatment with gold nanomaterials disclosed herein or compositions thereof or pharmaceutical compositions comprising the above ingredients according to the present application.

Pharmaceutical preparation

The gold nanomaterials of the invention are formulated with conventional carriers and excipients, which will be selected in accordance with conventional practice. Tablets will contain excipients, glidants, fillers, binders and the like. Aqueous formulations are prepared in sterile form and are intended to be generally isotonic by delivery other than orally.

While the active ingredients gold nanomaterials may be administered alone, it may be preferable to present them as pharmaceutical formulations. The formulation of the invention for veterinary and human use comprises at least one active ingredient as defined above (said active ingredient being gold nanomaterial, the same applies hereinafter) together with one or more acceptable carriers and optionally other therapeutic ingredients such as corticosteroids, anti-inflammatory signal transduction modulators, β 2-adrenoceptor agonists bronchodilators, anticholinergics, mucolytics, hypertonic saline and other drugs for the treatment of infections with viruses of the coronavirus family; or mixtures thereof.

Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. Or prepared into oral administration preparations by other known techniques.

Tablets are prepared by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant or dispersing agent. Molded tablets may be prepared by molding in a suitable machine a mixture of the powdered active ingredient moistened with an inert liquid diluent. The tablets may optionally be coated or scored and optionally formulated to provide slow or controlled release of the active ingredient therefrom.

The pharmaceutical composition of the present invention may be an external preparation such as an ointment for topical administration. For infections of the eye or other external tissues such as the mouth and skin, the formulation is preferably as a topical ointment or cream containing the active ingredient. When formulated as an ointment, the active ingredient may be used with either a paraffin or water-miscible ointment base. Alternatively, the active ingredient may be formulated in a cream with an oil-in-water cream base. If desired, the topical formulations may include compounds that enhance the absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such skin penetration enhancers include dimethyl sulfoxide and related analogs. The oil phase of the emulsions of the invention may be constituted from known ingredients in a known manner. The oily phase may comprise only emulsifiers, but it may also comprise mixtures of at least one emulsifier with a fat or an oil or with both a fat and an oil. Preferably, a hydrophilic emulsifier is included with a lipophilic emulsifier that acts as a stabilizer. Oils and fats are also preferably included.

The pharmaceutical compositions of the present invention may be in the form of a sterile injectable preparation, for example, a sterile injectable aqueous or oleaginous suspension.

The pharmaceutical compositions of the present invention may be in formulations suitable for topical administration to the eye including also eye drops, wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient.

The pharmaceutical compositions of the invention may be in the form of preparations suitable for topical administration in the oral cavity including lozenges comprising the active ingredient in a flavoured base, usually sucrose and acacia or tragacanth; lozenges comprising the active ingredient in an inert base such as gelatin and glycerol or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

The pharmaceutical compositions of the invention may be formulations for rectal administration which may be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate.

The pharmaceutical compositions of the present invention may be formulations suitable for intrapulmonary or intranasal administration, such formulations typically having a particle size in the range of 0.1 to 500 microns, such as 0.5, 1, 30, 35 microns and the like, which are administered by rapid inhalation through the nasal passage or by oral inhalation to reach the alveoli. The active ingredients of the present invention have a size that can be used for intrapulmonary or intranasal administration, and can be used for intrapulmonary or intranasal administration, such as inhalants.

The pharmaceutical compositions of the present invention may be in the form of preparations suitable for parenteral administration including aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may contain suspending agents and thickening agents.

Formulations of the pharmaceutical compositions of the present invention are presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit dosage formulations are those containing a daily dose or unit daily sub-dose, as defined above, or an appropriate fraction thereof, of the active ingredient.

The pharmaceutical composition of the invention may be a veterinary composition comprising at least one active ingredient as defined above and a veterinary carrier.

The compounds of the present invention are useful for providing controlled release pharmaceutical formulations containing as an active ingredient one or more of the active ingredients of the present invention, wherein the release of the active ingredient is controlled and modulated to allow less frequent administration or to improve the pharmacokinetic or toxicity profile of a given active ingredient.

Combination therapy

The medicament, pharmaceutical composition or active ingredient of the present invention is also used in combination with other active ingredients. For the treatment of coronavirus infection, the other active ingredients are active against coronavirus infection, in particular SARS or SARS-CoV-2 infection. Non-limiting examples of such other active therapeutic agents are corticosteroids, anti-inflammatory signal transduction modulators, β 2-adrenoceptor agonists bronchodilators, anticholinergics, mucolytics, hypertonic saline, and other drugs used to treat infections with viruses of the coronavirus family; or mixtures thereof.

The medicaments, pharmaceutical compositions or active ingredients of the present invention may also be administered to a patient simultaneously or sequentially in combination with one or more other active ingredients in a unit dosage form. The combination therapy may be administered as a simultaneous or sequential regimen. When administered sequentially, the combination may be given in two or more administrations.

Co-administration of a drug, pharmaceutical composition or active ingredient of the invention with one or more other active ingredients generally refers to the simultaneous or sequential administration of a compound of the invention and one or more other active therapeutic agents such that a therapeutically effective amount of both the drug, pharmaceutical composition or active ingredient of the invention and the one or more other active ingredients are present in the patient.

Combination therapy can provide "synergy" and "synergy", i.e., the effect obtained when used together is greater than the sum of the effects produced by the compounds used alone. When the active ingredients are: (1) When co-formulated and administered or delivered simultaneously in a combined preparation; (2) alternating or parallel delivery as separate formulations; or (3) by other schemes, a synergistic effect can be obtained.

One application scheme of the invention provides the application of the gold nano material in preparing a material for inhibiting coronavirus infection.

In one application scheme, the invention provides a material for inhibiting coronavirus infection, and the material comprises gold nano-materials and a matrix.

In the scheme of the material for inhibiting coronavirus, the material can also comprise a substrate which can be changed according to the application, and the gold nano-material has high stability and can adapt to various substrates, so that the material of the corresponding substrate can be obtained according to the conventional method in the field, including but not limited to the following steps of compounding the gold nano-material with the substrate, and adsorbing the gold nano-material with the surface of the substrate in an adsorption mode; or the gold nanometer material and the matrix are blended, and then the matrix is solidified to obtain the gold nanometer material.

In the technical scheme of the invention, in the material, the material is a disinfectant.

In a specific preferred embodiment, the material for inhibiting coronavirus infection is a material for preparing a mask, and the material is a fabric, and the surface or the inside of the fabric contains the gold nano-material. In a variable preferred embodiment, the material for inhibiting coronavirus infection is a material for preparing a mask, and the material is a non-woven fabric, preferably a spun-bonded non-woven fabric or a melt-blown non-woven fabric; the surface or the inside of the non-woven fabric contains the gold nano material. The preparation method of the material can adopt the conventional method in the field to compound the gold nanometer material and the matrix material. For example, the meltblown material may be combined with the gold nanomaterial before meltblown spinning, and then meltblown, or the obtained meltblown may be combined with the gold nanomaterial.

In a particularly preferred embodiment, the material inhibiting coronavirus infection is a coating which is a suspension comprising gold nanoparticles. Preferably, the coating further comprises at least one of a surfactant and a thickener. The coating is capable of imparting gold nanomaterials to surfaces of objects, such as packaging surfaces, medical device surfaces, cosmetic device surfaces, and the like, such that it forms a coating comprising the gold nanomaterials on the surfaces of the objects. In a specific embodiment, the coating is used for endowing the surface of the food or medicine outer package or inner package with a coating layer of gold nano-material, and preferably, the food or medicine needs to be transported in a cold chain.

In a particularly preferred embodiment, the material inhibiting coronavirus infection is a disinfectant, which is a suspension comprising gold nanoparticles. In a particularly preferred embodiment, the disinfectant is an environmental disinfectant, a leave-on hand sanitizer, a detergent. In a particularly preferred embodiment, the disinfectant is a surface disinfectant that can be used for packaging materials, medical instruments (e.g., catheters, injection needles, surgical instruments, surgical masks, and other medical devices), oral instruments (e.g., dentures, protective strips, padding, palatal expanders), cosmetic instruments (e.g., cosmetic instruments, orthopedic devices). In a particularly preferred embodiment, the concentration of gold nanomaterial in the disinfectant is from 1 μ g/mL to 1000 μ g/mL, preferably from 5 μ g/mL to 240 μ g/mL, more preferably from 7.5 μ g/mL to 120 μ g/mL.

In a particularly preferred embodiment, the material inhibiting coronavirus infection is a packaging material, and the packaging material is used for medicines or foods for cold chain transportation or storage. The surface of the packaging material is provided with a coating containing gold nano-materials, or the packaging material is a composite material containing gold nano-materials.

In a specific preferred embodiment, the material for inhibiting coronavirus infection is a filter material, and the filter material is a material for air filtration, a material for water body filtration or a material for mask filtration. In a specific preferred embodiment, the filtering material is an air filtering net, and the surface of the air filtering net is provided with gold nano-materials. In a specific preferred embodiment, the filtering material is air filtering cotton, and gold nano-materials are attached to the fibers of the air filtering cotton. For example, the filter material can be used for air conditioner filtration, air purifier filtration, fresh air system filtration and any other devices or equipment needing air filtration. The scene of use is not limited to homes, offices, laboratories, factories, etc.

In an embodiment of the present invention, in the above application, the material is a solid, liquid or semisolid material.

In the technical solution of the present invention, in the above-mentioned use, the liquid material is a paint containing a gold nanomaterial, a disinfectant containing a gold nanomaterial, or a personal care product containing a gold nanomaterial.

In the present invention, in the above-described use, the solid material is a device or a packaging material containing gold nanomaterial on the surface.

In an embodiment of the present invention, in the above application, the semisolid material is a gel containing a gold nanomaterial.

In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof are described in detail below, but the present invention is not to be construed as being limited to the implementable range thereof.

Experimental Material

ACE2/293T and Vero-E6 cells, pseudovirus SC2-P. Materials, reagents, carriers, strains and the like used in the following examples are commercially available unless otherwise specified.

The HEK-293T cell is a derivative strain of human embryonic kidney cell 293, is a cell line derived from human embryonic kidney cell, has the characteristics of high transfection efficiency, easy culture and the like, is a very common cell line for expressing and researching exogenous genes, and becomes a powerful tool for researching gene functions of extensive researchers.

ACE2/293T, refers to the expression of ACE2 in HEK-293T cells.

ACE2 is also known as achh and is called angiotensin converting enzyme 2. The protein coded by the gene belongs to an angiotensin converting enzyme family of dipeptidyl carboxyl dipeptidase and has considerable homology with human angiotensin converting enzyme 1. The gene coded protein is the functional receptor of SARS and SARS-CoV-2, HCoV-NL63 human coronavirus Spike glycoprotein.

The Vero-E6 cell is an African green monkey kidney cell line, is one of the verified cells capable of being infected by the new coronavirus, and can be used as a cell host for culturing the new coronavirus. Such as: the effect of a drug on the rate of replication of new coronaviruses is determined, and the presence of new coronaviruses is examined or cultured for research purposes.

The pseudovirus SC2-P is pseudovirion of SARS-CoV-2, which comprises marker genes such as Luciferase/GFP coated with Spike protein, which simulates virus-infected cells, and which is used for detecting the expression of the marker genes, and the SC2-P used in the following examples can be purchased from Jinwei corporation.

SARS-P is a pseudovirus of SARS and is a commercial product, and will not be described herein.

The gold nanomaterial (Au) used in the examples was nanoparticles having a particle size of less than 100 nm.

Example 1 detection of physicochemical characterization of gold (Au) nanomaterials

The particle size DLS detection and transmission electron microscope observation are carried out on the Au nano particles, the experimental result is shown in figure 1, and the particle size is about 5nm.

Example 2 detection of influence of Au nanomaterial on infection of ACE2/293T cells by SC2-P

ACE2/293T cells were seeded into 96-well plates at a density of 1X 10 ⁴ cell/wall，37℃，5％CO ₂ The culture is carried out overnight,

respectively infecting ACE2/293T cells for 2h by SC2-P mixed with 0/30/60/90 mu g/mL Au nano materials, washing once by DMEM, adding a DMED culture medium, and culturing for 40-48h, and then detecting the infection efficiency.

And (3) test results:

the results of luciferase activity assays showed that SC2-P was able to integrate the genome into the host cells (fig. 2). When Au nanomaterial treatment was applied, the amount of SC2-P infestation was significantly reduced compared to the control (fig. 2). The Au nano material can effectively inhibit the infection of SC2-P to host cells.

Example 3 examination of the Effect of Au nanomaterials on SARS-CoV-2 infection of Vero-E6 cells

Vero-E6 cells were seeded at a density of 5X 10 in 24-well plates ⁴ cell/wall，37℃，5％CO ₂ The culture is carried out overnight,

Vero-E6 cells are infected by a euvirus SARS-CoV-2 mixed with 0/7.5/15/60/120 mu g/mL Au nano material for 2h, DMEM is used for washing once and then added with a DMED culture medium for culturing for 40-48h, and then infection efficiency is detected. After the infected Vero-E6 cells are harvested, RNA is extracted, and the mRNA levels of the nonstructural protein (NSP 2) and the spike protein (S protein, spike) in the host cells are respectively detected by quantitative PCR.

And (3) test results:

the experimental results show that the addition of Au nano-materials can effectively reduce the mRNA level of non-structural protein (NSP 2) and spike protein (S protein, spike) in host cells, which indicates that the Au nano-materials can reduce the infection efficiency of SARS-CoV-2 (figure 3). This shows that Au nano-material can not only effectively inhibit the infection of SC2-P to host cells, but also inhibit the infection of virus SARS-CoV-2. As SC2-P is the simulation of SARS-CoV-2 virus surface S protein, according to the experimental result, the gold nano material can be expected to competitively act on ARS-CoV-2 virus surface S protein to inhibit the combination of the ARS-CoV-2 virus surface S protein and the host cell receptor ACE2.

Claims

1. The application of gold nanometer material in preparing medicine for preventing and treating coronary virus caused diseases, wherein the coronary virus is SARS-CoV2; the disease caused by coronavirus is a disease caused by infection of coronavirus SARS-CoV 2.

2. The gold nanometer material is used for preparing drugs or preparations for inhibiting coronavirus from infecting host cells of the coronavirus or preparing nonstructural proteins or spike proteins of the coronavirus, wherein the coronavirus is SARS-CoV2; the disease caused by coronavirus is a disease caused by infection of coronavirus SARS-CoV 2.

3. The gold nanometer material is used for preparing a material for inhibiting infection of coronavirus or reducing and killing the coronavirus, wherein the coronavirus is SARS-CoV2; the disease caused by coronavirus SARS-CoV2 is a disease caused by coronavirus infection.