CN111184708A

CN111184708A - Application of silver salt of fumaric acid monoethyl ester in resisting novel coronavirus infection

Info

Publication number: CN111184708A
Application number: CN202010114378.XA
Authority: CN
Inventors: 刘威衢; 刘威鑫; 胡剑; 蓝和彪; 刘威玲
Original assignee: Ganzhou Anhong Environmental Protection Technology Co ltd
Current assignee: Ganzhou Anhong Environmental Protection Technology Co ltd
Priority date: 2020-02-25
Filing date: 2020-02-25
Publication date: 2020-05-22

Abstract

The invention relates to application of silver fumarate monoethyl ester in resisting novel coronavirus infection. Adding 1000g of monoethyl fumarate into a 5L reaction bottle, adding 3 Kg of pure water, adding 100g of nano silver oxide under stirring, stirring for reaction for 9 hours, filtering the reaction solution to remove impurities, carrying out reduced pressure distillation at 60 ℃ to remove water, adding 5Kg of ethanol, stirring for crystallization for 4.5 hours, filtering, collecting solids, and carrying out vacuum drying on the obtained solids at 75 ℃ for 6 hours to obtain the monoethyl fumarate silver salt. Can be used for preventing and treating various coronavirus. Experiments show that: under the concentration of 500 mu g/mL, the inhibition rate of the fumaric acid monoethyl ester silver salt dissolving solution on main protease of coronavirus reaches 99.9 percent; the inhibition rates of the corresponding zinc salt solution and silver salt solution on the main protease are respectively 99.9% and 99.7%, and the effect is obvious.

Description

Application of silver salt of fumaric acid monoethyl ester in resisting novel coronavirus infection

Technical Field

The invention relates to the field of pharmacy, in particular to a monoethyl fumarate manganese salt, and especially relates to an application of a monoethyl fumarate silver salt in resisting novel coronavirus infection. Can be used for preventing and treating various coronavirus.

Background

The novel coronavirus is a special living body, has a strict living mode of living cell parasitism, and can parasitize in cells of human, animals and plants, insects, fungi, bacteria and the like to cause infection. In particular, SARS and new coronavirus can live and spread with indoor air bacteria. 75% of human infections are caused by viruses. One propagation cycle of a virus involves the entry of a host by recognition of host cell surface receptors or the injection of genetic material into the host (phage), the use of self nucleic acids as pathogen nucleic acids and protein molecules. SARS is a virulent infectious disease with a great threat to human beings, and no specific medicine or vaccine is available on the market at present.

Severe acute respiratory syndrome, also known as infectious novel coronavirus, is a serious acute respiratory infectious disease. The pathogen is a virus of the genus coronavirus. Coronaviruses belong to the family coronaviridae, which are positive-stranded RNA viruses, the genomes of which are largest among currently known positive-stranded RNA viruses. This genus contains about 26 species; this genus can be divided into three groups again, according to their natural host, gene sequence and serotype relationships: wherein the first group comprises transmissible gastroenteritis virus of swine, etc.; the second group has coronavirus, etc.; the third group includes avian infectious bronchitis virus and the like. The genome of coronavirus 2/3 to 3/4 encodes two replicase polyproteins which are responsible for the normal transcription and replication functions of the coronavirus main protease (main protease for short) only after the protease encoded by the virus is cleaved into independent subunits. If the hydrolysis of the main protease of the coronavirus can be inhibited, the coronavirus can be effectively prevented from infecting the human body. Thus, the main protease of coronaviruses is a major target for anti-RNA drug screening.

Inorganic and organic products, also known as secondary metabolites, have structural diversity and nanoactivity diversity. The organic nano-lipid product and the derivatives thereof play unlimited roles in the prior disease treatment and are one of the most potential resources in the current drug development process. At present, researches on traditional Chinese medicines such as plantain seed polysaccharide and the like, marine organisms and natural products in the microbial metabolism process are not in the way of being researched, a large number of compounds with novel structures are discovered every year, and the novel compounds are important sources of medicine lead compounds and play an important role in discovery of new medicines and lead compounds. The research on inorganic nano zinc and organic nano lipid products, particularly products derived from silver salts of monoethyl fumarate, manganese salts and the like, is necessary, and a huge treasury of the silver salts of monoethyl fumarate, the manganese salts of monoethyl fumarate and the small molecular compounds of plantago seed polysaccharide is necessary, so that the screening of important virus or important disease related target protein inhibitors from the silver salts of monoethyl fumarate, the manganese salts of monoethyl fumarate and the plantago seed polysaccharide products is necessary.

At present, no medicine specially aiming at SARS passes clinical experiments. Although SARS is effectively controlled worldwide, the human cannot guarantee that SARS storm will not be heavy enough, and in order to reduce the loss of life and property, it is important to develop highly effective anti-SARS and novel coronavirus medicines aiming at the existing research results.

Although Chinese herbal medicines have been used for treating various diseases, the application of silver salts of monoethyl fumarate and manganese salts of monoethyl fumarate and psyllium seed polysaccharide in virucidal inhibition of RNA coronavirus main protease activity has not been found.

Disclosure of Invention

In the process of researching the silver salt of monoethyl fumarate, the manganese salt of monoethyl fumarate and the plantain seed polysaccharide gum, the inventor finds that the combination of the silver salt of monoethyl fumarate, the manganese salt of monoethyl fumarate and the plantain seed polysaccharide gum has the effect of inhibiting the activity of RNA coronavirus main protease. It can be prepared into RNA coronavirus main protease inhibitor, thereby preventing or treating novel coronavirus infection.

The invention aims to provide application of a silver salt of monoethyl fumarate, a manganese salt of monoethyl fumarate and a psyllium seed polysaccharide gum in preparation of a medicament for preventing or treating novel coronavirus infection, in particular application of the silver salt of monoethyl fumarate in preparation of a novel coronavirus main protease inhibitor, and application of the silver salt of monoethyl fumarate in treatment and prevention of various coronaviruses.

The preparation method of the medicine for the application of the silver fumarate monoethyl ester in resisting the novel coronavirus infection comprises the following steps:

a. the preparation method of the monoethyl fumarate comprises the following steps:

adding 200g of maleic anhydride and 145mL of absolute ethyl alcohol into a 1000mL three-necked bottle, starting stirring, heating to 60 ℃ in a water bath, reacting for 3 hours, cooling to room temperature, adding 2 g of anhydrous aluminum chloride into the reaction bottle, heating to 90 ℃, reacting for 2 hours, adding 160mL of toluene, filtering while hot, stirring and cooling the filtrate to obtain a large amount of white solid, and filtering to obtain the monoethyl fumarate.

b. The preparation steps of the silver fumarate monoethyl ester are as follows:

adding 1000g of monoethyl fumarate into a 5L reaction bottle, adding 3 Kg of pure water, adding 100g of nano silver oxide under stirring, stirring for reaction for 9 hours, filtering the reaction solution to remove impurities, carrying out reduced pressure distillation at 60 ℃ to remove water, adding 5Kg of ethanol, stirring for crystallization for 4.5 hours, filtering, collecting solids, and carrying out vacuum drying on the obtained solids at 75 ℃ for 6 hours to obtain the monoethyl fumarate silver salt.

c. The preparation method of the fumaric acid monoethyl ester manganese salt comprises the following steps:

adding 1000g of monoethyl fumarate into a 5L reaction bottle, adding 3 Kg of pure water, adding 50g of manganese oxide while stirring, stirring for reacting for 8 hours, filtering the reaction liquid to remove impurities, carrying out reduced pressure distillation at 63 ℃ to remove water, adding 5Kg of ethanol, stirring for crystallizing for 5 hours, filtering, collecting solids, and carrying out vacuum drying on the obtained solids at 75 ℃ for 6 hours to obtain the monoethyl fumarate.

d. The preparation method of the plantain seed polysaccharide comprises the following steps:

(1) and (3) drying: placing semen plantaginis in a drying oven, adjusting temperature to 50 deg.C, and oven drying for 4.5 h. (2) polysaccharide preparation: removing impurities from semen plantaginis, pulverizing, breaking shells, and extracting with hot water for 3min at 90 deg.C for 3h at a weight/volume ratio (kg/L) of 1:30 for 3 times.

(3) Centrifuging the extracted polysaccharide leaching solution, controlling the centrifugal rotation speed at 4000 r/min, centrifuging for 10min, and firstly separating to obtain a supernatant; separating polysaccharide suspended on the residue from the residue, and drying the polysaccharide to obtain high viscosity semen plantaginis polysaccharide gum. (4) And (4) concentrating the supernatant obtained in the step (3), precipitating with ethanol until the final mass concentration of ethanol is 80%, centrifuging to obtain polysaccharide, and drying to obtain the low-viscosity plantain seed polysaccharide gum. The form of the preparation can be selected from various forms according to the purpose of treatment, and representative forms thereof include tablets, pills, powders, liquids, suspensions, capsules, suppositories, injections (liquids, suspensions, etc.), sprays, aerosols, inhalants, sustained-release microcapsules, and also a kit for antiviral masks and fumigation for dynamic killing of indoor air viruses, and application to anti-novel coronavirus agents.

The amount of the active ingredient contained in the pharmaceutical composition of the present invention can be suitably selected from a wide range, and is usually 000001 to 70% by weight, preferably 00001 to 6% by weight, and these can be further classified into oral agents, parenteral agents, nasal agents, vaginal agents, suppositories, sublingual agents, ointments and the like according to the administration route, and can be prepared, shaped and prepared by a usual method.

The method of administration of the pharmaceutical preparation is not particularly limited, and can be determined according to the form of the preparation, the age, sex, other conditions, and degree of disease of the patient. For example, tablets, pills, solutions, suspensions, emulsions, granules and capsules can be administered orally, i.e., injections can be administered singly or mixed with a usual auxiliary solution such as glucose and amino acids, and then intravenously, further, they can be administered singly in the muscle, intracutaneously, subcutaneously or intraperitoneally, if necessary, suppositories can be administered intrarectally, vaginal agents can be administered vaginally, nasal agents can be administered intranasally, and sublingual agents can be administered orally.

The dose of the pharmaceutical preparation is not particularly limited, and may be suitably selected from a wide range according to the desired therapeutic effect, administration method, treatment period, age, sex, other conditions of the patient, etc., and generally, the dose of the active ingredient administered per 1kg body weight per 1 day is about 001mg to 100mg, preferably 01mg to 50mg, for an adult, and the preparation may be administered 1 time per 1 day or divided into several times. In addition, in the treatment methods of the present invention, the anti-SARS agent may also be used in combination with, for example, other antiviral agents.

The invention also provides application of any one of maleic anhydride, semen plantaginis, methanol or ethanol, toluene and aluminum chloride extracts in preparing a medicament for inhibiting coronavirus infection, in particular application in preparing a novel coronavirus main protease inhibitor.

The beneficial effects of the application of resisting the novel coronavirus infection are as follows:

1. the invention adopts an in vitro activity measurement method to perform activity measurement on the silver salt of monoethyl fumarate, the manganese salt of monoethyl fumarate and the plantain seed polysaccharide gum, and the result shows that the silver salt of monoethyl fumarate, the manganese salt of monoethyl fumarate and the plantain seed polysaccharide gum have good effect of inhibiting the activity of main protease of coronavirus, thus indicating that the silver salt of monoethyl fumarate and the plantain seed polysaccharide gum can become potential drugs for preventing or treating novel coronavirus infection, or form a pharmaceutical composition with pharmaceutically acceptable proper carriers or excipients for preventing or treating novel coronavirus infection.

2. The key point is that the new invention of applying the silver salt of the monoethyl fumarate, the manganese salt of the monoethyl fumarate and the plantain seed polysaccharide gum to the resistance of the novel coronavirus infection is solved.

3. Breaks through the problem that no new medicine is available for treating the novel coronavirus infection.

Detailed Description

In order to better illustrate the present invention, specific embodiments thereof will be described in detail below.

Example 1

b. The preparation steps of the silver fumarate monoethyl ester are as follows:

(3) Centrifuging the extracted polysaccharide leaching solution, controlling the centrifugal rotation speed at 4000 r/min, centrifuging for 10min, and firstly separating to obtain a supernatant; separating polysaccharide suspended on the residue from the residue, and drying the polysaccharide to obtain high viscosity semen plantaginis polysaccharide gum. (4) And (4) concentrating the supernatant obtained in the step (3), precipitating with ethanol until the final mass concentration of ethanol is 80%, centrifuging to obtain polysaccharide, and drying to obtain the low-viscosity plantain seed polysaccharide gum. The form of the preparation can be selected from various forms according to the purpose of treatment, and representative forms thereof include tablets, pills, powders, liquids, suspensions, capsules, suppositories, injections (liquids, suspensions, etc.), sprays, aerosols, inhalants, sustained-release microcapsules, and supporting products for antiviral nasal masks and sustained-release dynamic killing of indoor air viruses, and can be applied to novel anti-coronavirus agents.

The dose of the pharmaceutical preparation is not particularly limited, and may be suitably selected from a wide range according to the desired therapeutic effect, administration method, treatment period, age, sex, other conditions of the patient, etc., and generally, the dose of the active ingredient administered per 1kg body weight per 1 day is about 001mg to 100mg, preferably 01mg to 60mg, for an adult, and the preparation may be administered 1 time per 1 day or divided into several times. In addition, in the treatment methods of the present invention, the anti-SARS agent may also be used in combination with, for example, other antiviral agents.

Pharmacological activity test part:

the specific method comprises the following steps:

1. the construction of the expression vector of the coronavirus main protease comprises the following specific steps:

a. performing in-vitro amplification by using a PCR (polymerase chain reaction) technology by using a cDNA library of a virus strain with the serial number of BJ01, which is provided by Beijing Hua large gene center;

forward guiding: 5'-CGGGATCCAGTGGTTTTAGGAAAATG-3'

And (3) reverse leading: 5'-CCGCTCGAGTCATTGGAAGGTAACACCAGA-3'

b. After the gene fragment amplified by PCR is cut by BamHI and XhoI double enzymes, the fragment with the size of about 1kb is recovered by agarose gel electrophoresis;

c. connecting the recovered fragment with a T vector, then transforming Escherichia coli DH5 α competent cells with the connection product, coating the competent cells on an LB plate (containing 100mg/L ampicillin), and culturing overnight;

d. a plurality of single colonies were picked up from the plate, inoculated into tubes containing about 5mL of LB (ampicillin was added to the LB solution so that the final concentration was 100mg/L), and cultured overnight. Then extracting plasmids by using a plasmid extraction kit, carrying out enzyme digestion by using BamHI and XhoI, and then recovering a target gene fragment with the size of about 1kb by using agarose gel;

e. the target vector pGEX-4T-1 (purchased from Pharmacia) was digested with BamHI and XhoI, and then the digested fragment was recovered by agarose gel;

f. and d, connecting the fragments obtained by the step e (mixing the target gene fragment and the target vector fragment after enzyme digestion recovery according to the molar ratio of 3: 1-6: 1, reacting for 30 minutes-18 hours at 16 ℃ according to the requirement of Takara DNA Ligation), transforming enterobacter coli (Escherichia coli) DH5 α competent cells, coating the competent cells on an LB (LB) plate (containing 100mg/L ampicillin) for overnight culture, screening positive clones for identification and sequencing, wherein sequencing results show that the coding gene of the main protease of the coronavirus is correctly cloned into a pGEX-4T-1 vector.

1.2 expression and purification of SARS coronavirus main protease, which comprises the following steps:

a. transforming the vector containing the gene pGEX-4T-1 encoding the coronavirus main protease obtained in the step 1.1 into a strain of Escherichia coli BL21(DE3), and screening positive clones by using an LB plate (containing 100mg/L of ampicillin);

b. selecting positive clones (single clones grown on LB plate containing ampicillin) on LB plate described in a, culturing overnight, transferring to 1L LB medium (containing 100mg/L ampicillin), adding 1mM IPTG when OD600 reaches 0.6-0.8, culturing at 16 deg.C for 12 hr;

centrifuging at 5000-8000 rpm for 10-15 min to collect cells, and then ultrasonically breaking the bacteria in ice bath for 20-30 min; centrifuging the bacterium breaking solution at 13000-15000 rpm for 20-40 min, and collecting the supernatant;

d. and adding the supernatant into a GST affinity chromatography column pre-equilibrated by PBS, and eluting 20-30 column bed volumes by PBS to remove the foreign proteins. Finally, adding about 2mL0.1mg/mL of human rhinovirus 3C protease, carrying out enzyme digestion at 4 ℃ for 12-20 hours, and then collecting coronavirus main protease;

e. and (d) purifying the coronavirus main protease obtained in the step d by using Mono Q (GE) anion exchange chromatography to obtain the coronavirus main protease with higher purity.

2. Screening method of coronavirus main protease inhibitor

The method for screening the coronavirus main protease inhibitor adopted by the invention is a screening method disclosed in CN101418334A, and the like, and comprises the following specific steps:

the activity of coronavirus main protease is determined by using fluorescent substrate MCA-AVLQSGFR-Lys (Dnp) -Lys-NH2 (purity is more than 95%, the amino acid sequence of the fluorescent substrate is derived from the N-terminal self-cleavage sequence of coronavirus main protease.

The instrument used for the fluorescence intensity measurement was a Fluoraskan Ascent fluorometer (ThermoLabsystems, Helsinki, Finland), and the wavelengths of the excitation light and the emission light were 320nm and 405nm, respectively.

Adding coronavirus main protease (final concentration: 0.5. mu.M) into a buffer solution (50mM Tris-HCl (pH 7.3), 1mM EDTA (with or without DTT)), adding DMSO lysate of a candidate sample (final concentration: 500. mu.g/mL, substrate concentration 20. mu.M, and 298K, standing for 10 minutes, rapidly adding a fluorescence-labeled substrate (MCA-AVLQSGFRL(DNP) L-NH2, final concentration 20. mu.M), setting a negative control, i.e., adding no candidate sample, and the rest conditions are the same, wherein excitation wavelength and emission wavelength are 320nm and 405nm, respectively, keeping the temperature at 298K, recording fluorescence reading every 3 seconds, determining 10 points in total, plotting by using time as an X axis and fluorescence value as a Y axis to obtain an enzyme activity mechanical curve, calculating the slope from the values of the first two points on the graph to obtain the initial speed V of the reaction, the initial reaction speed of the negative control is defined as V0, the initial reaction speed of the candidate sample is defined as Vi, so as to calculate the residual activity of the main protease after the corresponding candidate sample is added (Vi/V0), the inhibition rate of the corresponding candidate sample is (1-Vi/V0), and the candidate sample with the residual activity less than 20% (or the inhibition rate more than 80%) can be further separated and purified to find more concentrated active sites.

The following examples may provide those skilled in the art with a more complete understanding of the present invention, but do not limit the invention in any way:

adding 1000ml of distilled water into 5g of the silver fumarate monoethyl ester and 50g of the plantain seed polysaccharide gum obtained in the embodiments b and d, and stirring and dissolving to obtain a solution for later use;

adding 1000ml of distilled water into 5g of the fumaric acid monoethyl manganese salt and 50g of the plantain seed polysaccharide gum in the embodiment c, and stirring and dissolving to obtain a solution for later use;

example b determination of the inhibitory activity of silver monoethyl fumarate salt and manganese monoethyl fumarate salt on coronavirus main protease;

the silver fumarate salt solution obtained in example b was prepared so that the concentration thereof was 50 mg/mL.

Adding coronavirus main protease (final concentration: 0.5. mu.M) to a buffer solution (50mM Tris-HCl (pH7.1)1mM EDTA (with or without DTT)), adding a solution of the above 2 samples (final concentration: 500. mu.g/mL, substrate concentration: 20. mu.M, 298K, after standing for 10 minutes, rapidly adding a fluorescence-labeled substrate (MCA-AVLQSGFRL(DNP) L-NH2, final concentration: 20. mu.M), setting a negative control, i.e., adding no monoethyl fumarate silver salt solution sample, the same conditions, excitation and emission wavelengths of 320nm and 405nm, respectively, keeping the temperature at 298K, recording fluorescence readings every 3 seconds, determining 10 points in total, plotting the time as X-axis and the fluorescence as Y-axis to obtain an activity inhibition curve of the 2 monoethyl fumarate silver salt solutions against coronavirus main protease, the following results were calculated from the curves: under the final concentration of 500 mu g/mL, the inhibition rate of the silver fumarate monoethyl ester solution on the main protease reaches 99.9 percent; the inhibition rates of the corresponding gingival salt solution and manganese salt solution on the main protease are respectively 99.9 percent and 99.7 percent.

According to the above examples, it can be seen that the 0.5% aqueous solution of silver salt of monoethyl fumarate and its 2 drugs all have good inhibitory activity against coronavirus main protease. The silver salt of monoethyl fumarate and manganese salt of monoethyl fumarate can be respectively applied to preparation of coronavirus main protease inhibitor, thereby preventing or treating coronavirus infection.

Embodiments of the present invention will be understood by those of ordinary skill in the art to which the present invention pertains that numerous changes, modifications, substitutions and alterations may be made without departing from the principles and spirit of the invention, and it is therefore intended that the foregoing description be regarded as illustrative rather than limiting.

Claims

1. Use of a silver salt of monoethyl fumarate for the preparation of a medicament for the prevention and treatment of coronavirus infection, characterised in that the silver salt of monoethyl fumarate is used for the preparation of a coronavirus main protease inhibitor.

2. The use of claim 1, wherein the silver salt of monoethyl fumarate, manganese salt of monoethyl fumarate, psyllium polysaccharide gum drug is prepared by the steps of;

a. preparation of monoethyl fumarate:

adding 200g of maleic anhydride and 145mL of absolute ethyl alcohol into a 1000mL three-necked bottle, starting stirring, heating to 60 ℃ in a water bath, reacting for 3 hours, cooling to room temperature, adding 2 g of anhydrous aluminum chloride into the reaction bottle, heating to 90 ℃, reacting for 2 hours, adding 160mL of toluene, filtering while hot, stirring and cooling the filtrate to obtain a large amount of white solid, and filtering to obtain monoethyl fumarate; b. the preparation steps of the silver fumarate monoethyl ester are as follows:

adding 1000g of monoethyl fumarate into a 5L reaction bottle, adding 3 Kg of pure water, adding 100g of nano silver oxide under stirring, stirring for reaction for 9 hours, filtering the reaction solution to remove impurities, carrying out reduced pressure distillation at 60 ℃ to remove water, adding 5Kg of ethanol, stirring for crystallization for 4.5 hours, filtering, collecting solids, and carrying out vacuum drying on the obtained solids at 75 ℃ for 6 hours to obtain a monoethyl fumarate silver salt;

adding 1000g of monoethyl fumarate into a 5L reaction bottle, adding 3 Kg of pure water, adding 50g of manganese oxide while stirring, stirring for reacting for 8 hours, filtering the reaction solution to remove impurities, carrying out reduced pressure distillation at 63 ℃ to remove water, adding 5Kg of ethanol, stirring for crystallizing for 5 hours, filtering, collecting solids, and carrying out vacuum drying on the obtained solids at 75 ℃ for 6 hours to obtain monoethyl fumarate;

d. the preparation method of the plantain seed polysaccharide gum comprises the following steps:

(1) and (3) drying: placing semen plantaginis in a drying oven, adjusting temperature to 50 deg.C, and oven drying for 4.5 h;

(2) polysaccharide preparation: removing impurities from semen plantaginis, pulverizing, breaking shells, and extracting with hot water for 3min at 90 deg.C for 3h at a weight/volume ratio (kg/L) of 1:30 for 3 times;

(3) centrifuging the extracted polysaccharide leaching solution, controlling the centrifugal rotation speed at 4000 r/min, centrifuging for 10min, and firstly separating to obtain a supernatant; separating polysaccharide suspended on the residue from the residue, and drying the polysaccharide to obtain high-viscosity semen plantaginis polysaccharide gum;

(4) and (4) concentrating the supernatant obtained in the step (3), precipitating with ethanol until the final mass concentration of ethanol is 80%, centrifuging to obtain polysaccharide, and drying to obtain the low-viscosity plantain seed polysaccharide gum.

3. Use according to claim 1, wherein the silver salt of monoethyl fumarate and the manganese salt of monoethyl fumarate are obtained by means of a solvent.

4. Use according to claim 2, wherein the solvent is selected from water, alcohols.

5. The method according to claim 2, wherein the amount of the active ingredient contained in the silver monoethyl ester fumarate and manganese monoethyl fumarate is appropriately selected from a wide range, and is usually 000001 to 70 wt%, preferably 00001 to 6 wt%, and these are further classified into oral agents, non-oral agents, nasal agents, vaginal agents, suppositories, sublingual agents, and ointments according to the administration route, and each of them can be blended, shaped, and prepared according to a usual method.

6. The use according to any one of claims 1 to 5, wherein the silver salt of monoethyl fumarate, manganese salt of monoethyl fumarate and psyllium polysaccharide gum are used for preparing coronavirus main protease inhibitors, and are used in the form of tablets, pills, powders, solutions, suspensions, capsules, suppositories, injection solutions, suspensions, sprays, aerosols, inhalants, sustained-release microcapsules, and kits for anti-viral nasal masks and sustained-release dynamic killing of room air viruses.