KR20060079388A - The synthesis of nano inorganic antiviral agent that is effective against sars corona virus and influenza virus - Google Patents

Abstract

The present invention relates to a method for synthesizing a new antimicrobial agent having antiviral ability against SARS-causing viruses and the like using a nano-sized solution of silver particles and a photocatalyst titanium dioxide (TiO ₂ ). More specifically, a solution of nano-sized silver particles (1-100 nm) colloid having antibacterial and bactericidal properties and a solution of titanium dioxide particles (1-100 nm), which is also a nano-sized photocatalyst, in an appropriate ratio and concentration, includes water containing alcohol and distilled water. And an inorganic antibacterial solution synthesis technology prepared by synthesizing with one or more solvents selected from the group consisting of surfactants and diluting them to be sprayable and to have bactericidal and antibacterial effects against various viruses, bacteria and fungi.

Inorganic antimicrobial materials synthesized with photocatalysts such as titanium dioxide and nano silver particles, alcohols, distilled water, and surfactants can exhibit a synergistic effect that enhances strong bactericidal and antiviral properties in the presence or absence of light. It was made. In addition, the inorganic antimicrobial material synthesized according to the present invention also has a function of removing pollutants and deodorizing function and fragrance function by added fragrance component by decomposing volatile organic and odorous substances by specific oxidation ability, and also being used as a spray type. There are possible features.

Inorganic antimicrobial solution synthesis technology according to the present invention is a new inorganic antimicrobial material manufacturing synthesis technology that overcomes the disadvantage of the limited manufacturing technology, the existing inorganic antimicrobial material can be manufactured only by mixing (Mix) or blending (Blending) technology There is a difference.

Nano, silver, titanium dioxide, photocatalysts, antibacterial agents, fungicides, sprays, SARS, coronaviruses, influenza viruses, synthesis

Description

Synthesis of nano inorganic antiviral agent that is effective against SARS Corona virus and Influenza virus}

Figure 1 is a TEM result photo of the new synthetic solution prepared by the present invention

The present invention is an inorganic antiviral and antimicrobial agent prepared by preparing a nano-sized silver particle colloid having a suitable concentration, synthesis of a titanium dioxide photocatalyst particle solution using the prepared colloidal solution as a solvent of a constant concentration in a sprayable form It relates to a new synthetic preparation method of. Products according to the invention are potent / sterilized against various viruses, bacteria and fungi, including the variant coronavirus, which is a cause of SARS that may be present in the air or on the surface of a human body, animal or object by spraying it into the air. And antiviral activity. In addition, it has an annihilation effect of 99.9% or more on the swine infectious gastroenteritis virus (TGEV) and the porcine epidemic diarrhea virus (PEDV), which are the causative agents of swine infectious gastroenteritis. In addition, the synthesized inorganic antibacterial substance has the effect of removing and deodorizing volatile organic substances or odorous substances by harmless carbon dioxide and water by powerful oxidation.

Silver has long been known to have a bactericidal effect. Particularly, silver colloid refers to a substance dispersed in an aqueous solution in a clustered state (10-150 nm in size). It has been found to have strong bactericidal power against harmful bacteria and fungi. In addition, it has been observed that these silver colloidal particles do not kill most beneficial bacteria when applied to the human body, and in the United States, silver colloidal solution is recognized as a harmless natural product to the human body that does not fall within the FDA pharmaceutical range, and recently it is a non-toxic preservative. As a food additive, it is known as a material that is harmless to humans or living bodies. As described above, the silver colloidal solution has been proved to be very sterilizable and non-toxic to humans. Currently, in many countries around the world, silver particles are more effective for sterilization, eradication and treatment of pathogenic bacteria using the same. Research has been conducted to utilize silver in various products such as clothing and clothing.

The photocatalyst refers to a material having an electrically semiconducting property that generates active oxygen and hydroxyl radicals (OH radicals) upon receiving light having an appropriate energy or more and exhibits decomposition and antibacterial properties of odorous substances by their strong redox action. In semiconductors, when a certain area of energy is applied, electrons transfer energy from the valence band to the conduction band (excitation, 勵 起, quantum mechanical system), and transition from low to high steady state. )do. At this time, electrons (e−) are formed in the conduction band and holes (h +) are formed in the valence band. The electrons and holes thus formed cause various reactions such as decomposing harmful substances by strong oxidation or reduction. Titanium dioxide (TiO ₂ ) is the most widely used among various oxide materials having properties as photocatalysts due to its chemical stability and excellent advantages as semiconductors. In the case of titanium dioxide, when it receives an energy of about 3.0 eV (equivalent to 380 nm wavelength in the energy gap between the valence band and the conduction band), it fully functions as a photocatalyst. E contact with the light generated in the photocatalyst is titanium dioxide (e-) and holes (h +) is a superoxide anion on the titanium dioxide surface causes the O _2, H ₂ O and the reaction of each air (O ₂ ^-) and hydroxyl radical ( To produce two free radicals, OH). In particular, since radicals have a high oxidation and reduction potential, they are excellent for NOx, volatile organic compounds (VOCs) and various odors, and are used for BOD, color and hardly degradable contaminants, environmental hormones, etc. Not only can it be completely removed, it also has the ability to oxidize all target substances, such as pathogenic E. coli, Staphylococcus aureus, and 0-157, which kills more than 99% of various pathogens and bacteria. The titanium dioxide photocatalyst is also reacted by solar energy or fluorescent light, and is economical because it exhibits a permanent function by the cycle of 'fixing to an object → photolysis → regeneration'. In addition, the by-products after the reaction is a material that is harmless to the environment by water and CO ₂ , it can be applied to various fields.

SARS is likely to be a variant of the existing corona virus that causes colds, according to the World Health Organization. Corona virus has a high incidence of mutations unique to RNA viruses, and has a high recombination frequency because RNA polymerases tend to jump from place to place while replicating genomes. Corona virus causes respiratory symptoms such as colds when it is infected with humans, but it has not been a major problem since it is not high in risk. However, some animals, such as cows, dogs, pigs, and birds, have sometimes acted as deadly pathogens, which is why infectious disease experts estimate that the corona virus has been transformed into livestock, such as pigs and chickens, into human bodies, causing deadly diseases. . SARS spreads the virus as it moves into the respiratory tract after inhaling air droplets, which are protruding drops when the patient coughs, or touching objects (such as door handles or telephone computer keyboards) used by the patient.

Prior art investigation and analysis of the bactericidal / sterile material by coronavirus inhibitor, silver or titanium dioxide photocatalyst are as follows. Currently, products having inhibitory ability against coronaviruses are mainly substances extracted from synthetic organic matter or natural products (Korean Patent Laid-Open Patent Publication No. 2003-0063961 and Japanese Patent Laid-Open Publication No. 2000-44473), and are not inorganic antimicrobial products such as this patent. Antimicrobial products using silver colloid or titanium dioxide photocatalyst alone as inorganic materials are based on the above-mentioned theory. Various products such as silver coated clothing, antibacterial spray (Korea Patent Publication No. 2002-0008375), filter coating agent for air purifier, etc. When silver is used, there is almost no change in performance according to ambient conditions, but a product using only a photocatalyst has a big disadvantage in that it does not properly display performance in an environment in which light is insufficient. In addition, as the inorganic antimicrobial agent of the present patent, a combination of silver and titanium dioxide photocatalyst to produce the antimicrobial technology and the technology for the manufacturing method disclosed in Korea Patent Publication No. 1998-0007982 (Method for preparing inorganic antibacterial agent), Patent 2001-0057595 Coated photocatalyst production method) and special 2003-0037050 (titanium dioxide photocatalyst containing an antibacterial metal component and its production method).

In particular, 2003-0037050 is a complex process of preparing antimicrobial metals such as silver in colloidal form, mixing them with titanium peroxide, hydrothermal synthesis, and crystalline oxides, and coating their surfaces with a sol-gel method to microencapsulate them. Photocatalyst material containing antibacterial metal is made through the process, and special 1998-0007982 is mixed with phosphoric acid, silver nitrate, titanium dioxide powder and colloidal silica, and then dried and calcined to prepare carriers and at the same time to prepare inorganic antimicrobial agent supporting silver ions. Patent Application No. 2001-0057595 relates to a method for producing a fine powder photocatalyst in the form of applying silver to the photocatalyst by substituting silver for the photocatalyst. Therefore, these composites have a disadvantage in that it is difficult to control the substitution concentration of silver ions and undergo a complicated manufacturing process such as composite hydrothermal synthesis, intermetallic substitution, and calcination. The greatest manufacturing differentiation is that the present invention produces inorganic antimicrobial substances by a novel synthesis method that is completely different from the past dilution and blending preparations (especially 2004-0005512).

The sterilization mechanism of the inorganic antimicrobial material prepared by the new synthetic method can be largely represented in two ways. 1) function of asphyxiation or assassination while preventing viruses, bacteria, fungi, etc. from blocking the function of the absorption system (Coenzyme A), which is required for oxygen and digestive metabolism (nanoparticles bind to the virus and coagulate); Nanoparticles bind to proteins such as cell membranes and enzymes to inhibit cell energy metabolism. 2) Nanoparticles penetrate into the virus / bacteria body and destroy and destroy the organic structure of the cells; This nanocomposite breaks down the cell membrane and disturbs the cells by making strong and instantaneous bonds with microorganisms such as -SH, -COOH, and -OH groups.

The present invention does not undergo complex processes such as hydrothermal synthesis, intermetallic substitution, and firing of two nanomaterials, silver colloid and photocatalyst dioxide, which have unique bactericidal and antiviral properties even when they exist independently. By diluting a solution in which nano-sized silver particle colloid was homogeneously dispersed at an appropriate ratio, and using it as a solvent necessary for the synthesis of titanium dioxide photocatalyst, a method for newly synthesized inorganic antimicrobial material was developed. In addition, it exhibits synergy effect by silver and photocatalyst in the bactericidal and antiviral ability, and especially nano silver antimicrobial agent synthesized with silver colloid and photocatalyst particles or alcohol, distilled water, surfactant It can be used to show strong sterilization and antiviral activity. Inorganic antimicrobial agent according to the present invention, as well as the elimination of various viruses, bacteria, fungi and odors, as well as a strong bactericidal and antiviral ability against the coronavirus and influenza virus, which causes SARS, the swine infectious gastroenteritis virus, which is the cause of swine infectious gastroenteritis (Transmissible GastroEnteric Virus, TGEV) and Porcine Epidemic Diarrhea Virus (PEDV) have a purpose of having an killing effect of more than 99.9%.

When nano-sized particles are present in a high concentration in the solution, the particles are agglomerated with each other by the self-association reaction, and thus, it is difficult to maintain homogeneity due to precipitation due to an increase in particle diameter. Therefore, in order to eliminate this disadvantage in the preparation of the present inorganic antimicrobial agent, after preparing a nanoparticle colloid of a certain size of nanoparticles, one or more solvents are selected from the group consisting of water, alcohols and surfactants containing distilled water within a certain concentration range. Titanium dioxide photocatalyst was synthesized using as a raw material.

The inorganic antimicrobial agent prepared a silver colloidal solution containing 50,000 ppm of silver particles having an average particle size in the range of 1 to 100 nm in size and diluted with distilled water to a concentration of 10 to 5,000 ppm, the average particle size of 1 to 100 nm Anatase-type titanium dioxide photocatalyst particles having a range were used as a solvent for preparing an inorganic antimicrobial agent solution containing a concentration of 0.01 to 5 wt%. It is important to note that the initial concentration of the colloidal solution and the photocatalyst particles, not the final concentration after synthesizing and diluting the two substances, indicates the properties of the inorganic antimicrobial agent according to the present invention. Therefore, the silver particle concentration of 10 to 5,000 ppm and the photocatalyst concentration of 0.01 to 5 wt% can be sufficiently controlled within the limits of homogeneous dispersion without aggregation. The inorganic antimicrobial solution prepared by the new synthetic method formed a uniform particle size and uniform distribution of 20 nm or less as a result of analysis through the TEM photograph of FIG. 1. The inorganic antimicrobial agent thus synthesized can be sprayed by diluting with distilled water, alcohol, etc. according to the application target, and can be applied to antibacterial and anti-fouling products by spraying or applying to other substances by preparing and spraying in a sprayable form.

Efficacy of the antimicrobial and antiviral activity of the inorganic antimicrobial agent prepared by the present patent is effective against influenza-A virus, Escherichia coli and Staphylococcus as well as swine epidemic diarrheal virus and swine infectious gastroenteritis virus, which are a kind of coronavirus, as shown in the following examples. It was found to be very good. The following examples do not limit the efficacy of the inorganic antimicrobial agent according to the present invention to only four substances to be tested. In addition, the oral toxicity test and skin irritation test were performed as part of the safety test to verify that the material is harmless to humans or animals.

(Example 1) Test result of bactericidal power against E. coli of inorganic antibacterial solution

After spreading E. coli (Escherichia coli) to the medium, the solution of the inorganic antimicrobial solution according to the present invention diluted 10 to ³ times in distilled water is sparged three times (about 0.5 ml). The conditioned media after three fluorescent lamps are cultured for 24 hours in the installed incubator by determining the number of colony were irradiated with germicidal The result is an excellent sterilizing power against 10 ³ by the inorganic antibacterial agent E-coli in terms dilution fold as shown in Table 1 Indicated.

[Table 1] Results of bactericidal activity by inorganic antibacterial agents

Example 2 Results of Antimicrobial Activity against Staphylococcus Aureus of Inorganic Antimicrobial Solution

100%, as the contact time of the inorganic antibacterial agent Staphylococcus aureus was added After a test for the antimicrobial activity of the (Staphylococcus Aureus), the test fungi in the samples 10, ^3-fold diluted solution in Table 2 when treated for 1 hour and 3 hours It showed antimicrobial activity. The test method is as follows.

1. Five colonies of LB broth medium (Peptone 10g, yeast extract 5g, NaCl 10g, agar 15-20g / L) were inoculated with single colony and cultured overnight at 37 ° C.

2. Turbidity was adjusted to 0.5 by measuring O.D. (optical density) at 660nm.

3. The test solution adjusted to 0.5 was inoculated in 5 ml of LB broth for 100-fold dilution.

4. Sample processing is as follows.

Treatment Concentration: Samples were treated so that the final concentration was 10 ³ fold dilution.

Treatment time: Samples were incubated for 1 hour and 3 hours, diluted (100 times, 10,000 times), and 1000 μl was applied to LB plates (LB broth + agar 15 to 20 g / L) and overnight at 37 degrees. Incubated.

5. The antimicrobial activity was measured by comparing the colony number of the control group without the colony count.

Table 2 Antimicrobial Activity of Staphylococcus Aureus

Example 3 Antiviral Activity Test Results for Coronaviruses of Inorganic Antimicrobial Agents

As a target virus to measure antiviral activity against the inorganic antimicrobial agent according to the present invention, PEDV (Porcine Epidemic Diarrhea Virus) and TGEV (Transmissible GastroEnteric Virus) were selected as coronaviruses. . PEDV is a coronavirus that proliferates in the monkey kidney cell line Vero cells, such as the animal cell line known as SARS virus (VeroE6), and the VeroE6 cell line is a derivative cell line of Vero cell line. Test methods, data processing and evaluation methods are as follows.

1. Viro host cells were placed in each well of a 96 well plate in 2 × 10 ⁴ cells and cultured for 16 hours to completely cover the cells with a monolayer on the bottom of each well.

2. The antimicrobial solution was diluted 10 times in distilled water, and the virus solution diluted to a constant concentration was mixed and left at 4 ° C for 30 minutes, and then inoculated into each well of the plate in which each cell was cultured.

3. As a control, a culture medium containing no inorganic antimicrobial solution, a culture medium containing only virus, and a well containing only the inorganic antimicrobial solution and containing no virus were used.

4. Test 96 well plates were terminated after 40 hours and the cells in 96 well plates were fixed with 70% acetone solution and completely dried.

5. The assay was performed by staining the cell proteins remaining in each well with SRB (0.4% Sulforhodamine B in 1% acetic acid) and re-dissolving them. Afterwards, the absorbance of each well was measured using an EIA reader (96 well plate reader). The ratio of the treated group and the untreated control group was analyzed.

6. Each test was repeated three times.

7. Based on the absorbance values of the group not treated with virus (A), the group treated with only inorganic antimicrobial solution (B), the group treated with virus only (C), and the group treated with both virus and inorganic antimicrobial solution (D) The antiviral activity was calculated by the following equation, and the overall performance of the inorganic antimicrobial solution was evaluated.

% Antiviral activity = (D-C) / (B-C) x 100

After diluting the inorganic antimicrobial solution in distilled water in the range of 10 ² ~ 10 ³ , the results of testing the antiviral ability according to the above procedure are shown in Table 3 (Repeated three times each showed a similar type). Within the range, the cytotoxicity by the inorganic antimicrobial solution was hardly shown. If hayeoteul diluted inorganic antibacterial agent solution 10 times ² is the antiviral capacity of the PEDV and TGEV has were all exhibit greater than 90% virus inhibitory ability by 99.9%, 93.0% at 99%, was 99.9% or more, 10 ³ times each generally The higher the concentration of the inorganic antimicrobial solution, the higher the inhibitory activity against the virus.

[Table 3] Evaluation results for antiviral activity of inorganic antibacterial solution

In conclusion, as a result of testing the antiviral ability by diluting the inorganic antimicrobial agent according to the present invention in distilled water 10 ² ~ 10 ³ times, the cytotoxicity was not shown within this test range, the concentration of the inorganic antimicrobial agent increased. Accordingly, antiviral activity was also increased, and the highest inhibition rate within this test range was over 99.99%. Therefore, when the treatment time is increased or when the inorganic antimicrobial agent is diluted with low magnification, it is expected to show almost perfect effect as an inhibitor of corona virus infection.

Example 4 Safety Test Results for Oral Toxicity and Skin Irritation of Inorganic Antibacterial Solution

As part of the safety test for the inorganic antimicrobial agent according to the present invention, oral toxicity test and skin irritation test were conducted in accordance with Korean Food Safety Notice No. 1999-61. The results are as follows.

[Table 4] Evaluation results for the safety of the inorganic antimicrobial solution (G-Sol)

The test results of Table 4 are as shown in *, **.

*: General symptoms and autopsy findings related to administration of test substance were observed after 14 days of oral administration of test substance to rats at 5,000mg / kg BW. Was not recognized, no death animals were observed, and no significant weight changes were recognized. Therefore, the approximate lethal dose for G-Sol is estimated to be more than 5,000 mg / kg B.W.

**: The skin application of G-Sol in New Zealand White rabbits does not cause erythema, skin and edema, so the PIR (Primary Irritation Index) value, which is the primary skin irritation index, is calculated as "0.0" It was evaluated by (None Irritation).

Through the above results, it was confirmed that the inorganic antimicrobial solution was harmless to humans or animals.

As described above, the inorganic antimicrobial solution synthesis technology according to the present invention is a new inorganic antimicrobial material manufacturing technology that overcomes the limited disadvantage that the existing inorganic antimicrobial material could be manufactured only by mixing (Mix) or blending (Blending) technology. There is a big technical difference compared to manufacturing technology. It synthesizes a new inorganic antimicrobial material or makes it in a sprayable form using a nanosized silver particle and a photocatalyst solution of titanium dioxide nanoparticles using one or more solvents selected from the group consisting of alcohol, water including distilled water, and a surfactant. In the presence or absence of light, nano silver particles and photocatalyst particles are characterized by the ability to exert a powerful synergistic effect to enhance antiseptic and antiviral activity. In addition, the inorganic antimicrobial substance possesses the function of decomposing volatile organic substances and odorous substances by unique organic substance oxidizing ability and fragrance function by the included fragrance components, and is manufactured in a sprayable form and is easy to use. Due to the above characteristics, the present invention has a strong bactericidal and antibacterial and odor for various viruses, bacteria and fungi, including varietal coronavirus, swine epidemic diarrheal virus, swine infectious gastroenteritis virus, E. coli, influenza, MRSA, etc. It has a wide range of uses for deodorization of sexual substances, decomposition of trace volatile organic substances, and the purpose of simultaneously obtaining the purpose of fragrance. Specifically, spaces containing a lot of airborne bacteria such as hospital operating rooms, postpartum care centers, nursery facilities, kindergartens, offices, airplane cabins, airports, apartments, etc. It is effective to remove and block the infection path of viral and bacterial infectious diseases with strong sterilization / sterilization power by spraying inorganic antibacterial agent to the space used by the elderly and children and the breeding ground related to the pig farming industry. It is expected to be environmentally friendly and ultimately contribute to mitigating air pollution and improving public welfare by eliminating oxidizing effects and molds.

Claims (4)

After preparing nano-sized silver particle colloid, nano-sized titanium dioxide photocatalyst was synthesized using a solution mixed with one or more solvents selected from the group consisting of water, alcohols and surfactants containing distilled water within a predetermined concentration range. Inorganic antimicrobial solution prepared by the method and a substance diluted thereto. The method of claim 1, Silver particles have an average particle diameter ranging from 1 to 100 nm, and have an inorganic antimicrobial solution in a concentration range of 10 to 5,000 ppm in the inorganic antimicrobial agent. The method of claim 1, Titanium dioxide, a photocatalyst, is an anatase type and has an average particle diameter ranging from 1 to 100 nm, and an inorganic antibacterial solution having a concentration range of 0.01 to 5 wt% in an inorganic antibacterial agent. The method of claim 1, The prepared inorganic antimicrobial solution and the material to which the diluted substance is applied, impregnated, or impregnated are natural fibers, nonwoven fabrics, masks, filters, and the like, and the application form thereof is characterized by the form of solutions, vials, sprays, aerosols, and the like. Antimicrobial and anti-pest products.

Images