Environmentally friendly epidemic prevention composition capable of killing phospholipid enveloped viruses

Abstract

The present invention relates to an environmentally friendly epidemic prevention composition that has the function of killing phospholipid enveloped viruses, and contains 10-40% by weight of polyoxyethylene lauryl ether, 0.01-10% by weight of polyoxyethylene 2-ethylhexyl ether, 0.01-10% by weight of polyoxyethylene polyoxypropylene alkyl ether, and the remaining amount of solvent, based on a total of 100% by weight. The environmentally friendly epidemic prevention composition according to the present invention has a 100% SARS-CoV-2 virus removal rate in 1 minute, an effect of killing 100% highly pathogenic avian influenza viruses in 1 to 10 minutes, and an IC20 value of 100% for human epithelial keratinocyte cells (HaCaT), human bronchial epithelial cells (BEAS-2B), and human monocyte cells (THP-1) is 100%. 50 The value (%) is in the range of 0.003 to 0.0045, indicating that the compound has low cytotoxicity and antibacterial activity against both non-pathogenic and pathogenic strains.

Description

[Technical field]

[0001] The present invention relates to an environmentally friendly epidemic prevention composition having the function of killing phospholipid enveloped viruses, and more particularly, to an environmentally friendly epidemic prevention composition that can effectively remove various phospholipid enveloped viruses such as COVID-19, avian influenza, African swine fever, and severe acute respiratory syndrome. [Background technology]

[0002] Viruses have a very simple structure consisting of genetic material (RNA or DNA) surrounded by a protein shell (capsid). Some viruses have a lipid membrane in addition to the protein shell, and those with this lipid layer (lipid layer, lipid membrane) are enveloped viruses.

[0003] This lipid layer is present only in enveloped viruses, and is known to be inactivated and lose infectivity when exposed to organic solvents such as ether. The lipid membrane of enveloped viruses is said to play a major role in introducing their own genes into the host's genome. It is expected that inhibiting this mechanism will enable effective combating of various types of viruses without causing unnecessary side effects.

[0004] Alcohol-based cleaners, which are currently widely used as cleaning agents to remove such viruses, contain ethanol as their main ingredient, which exerts a disinfecting effect by dissolving the viral envelope (phospholipids), and are currently widely used in Korea.

[0005] Ethanol dissolves the envelope of enveloped viruses, which are made up of a phospholipid layer and proteins, and either kills non-enveloped viruses on the outside or prevents them from penetrating and replicating inside the host cell even if they are lucky enough to invade the host.

[0006] On the other hand, influenza viruses infect the respiratory system and cause systemic symptoms, and not only change their form periodically, but also spread to other hosts before the host dies without killing them, so scientists speculate that influenza viruses will survive until the extinction of humanity. Influenza viruses are the viruses that cause the greatest economic losses to humanity, and although preventive vaccines have been developed, they cannot keep up with the mutations of the virus and have not yet led to a fundamental treatment.

[0007] Among them, Avian Influenza (AI) is an acute infectious disease caused by infection with the avian influenza virus, and causes serious damage to poultry such as chickens, turkeys, and ducks. It is broadly classified into low pathogenic avian influenza and highly pathogenic avian influenza according to the degree of pathogenicity of the virus.

[0008] The four pandemic viruses that emerged between 1918 and 2009 were the Spanish flu (A / H1N1), which caused the most deaths, the Asian flu (A / H2N2), the Hong Kong flu (A / H3N2), and the swine flu (A / H1N1)pdm09, which killed 260 people in South Korea in 2009.

[0009] The outbreak of avian influenza viruses, which are most likely to cause the fifth pandemic, continues to be reported. Avian influenza A / H5N1, which was first discovered in 2003, broke out in wild birds and poultry and spread to humans, killing 454 people in 16 countries by September 2017. In addition, avian influenza A / H7N9, which broke out in China in 2013, has killed 599 people as of 2017.

[0010] Meanwhile, SARS (Severe Acute Respiratory Syndrom) is a novel infectious disease that spread throughout the world, including Hong Kong, Singapore, Canada, etc., within a few months of its outbreak in China in the winter of 2002. The causative pathogen is SARS-associated coronavirus (SARS-coV). SARS-coV is known to be capable of cross-species infection from animals to humans through a variant of an animal-host coronavirus.

[0011] The basic route of infection for SARS is known to be through respiratory droplets or direct or indirect contact with mucous membranes via contaminated media. Symptoms include fever, fatigue, muscle pain, headache, and chills, and it has been reported that the disease is mainly transmitted in the second week.

[0012] Patients suspected or suspected to have SARS are isolated and hospitalized, and strict management is required to prevent the spread of the virus. However, since no vaccine or preventive medicine has yet been developed, the only way to prevent infection through direct contact is to refrain from traveling to areas with a high risk of infection and to wash your hands thoroughly.

[0013] When the structure of coronavirus is observed with a transmission electron microscope (TEM), it is found to be composed of a double phospholipid membrane. 10 ~C 18 It has a structure with a long carbon chain.

[0014] The applicant disclosed in Korean Patent Application No. 2020-0137611 that an antiviral composition for preventing viruses is 13 H 27 (OCH2CH2) n The paper discloses a cleaning composition comprising 10-30% by weight of OH (wherein n=2-15), 0.1-5% by weight of 2-ethylhexyl alcohol ethoxylate, 0.1-5% by weight of an ethylene oxide-propylene oxide block copolymer, and the balance water.

[0015] C, which is the main component in the above patent 13 H 27 (OCH2CH2) n Since OH has a branched structure rather than a linear structure with 13 carbon chains, it can bind well to the hydrophobic portion of the coronavirus phospholipid bilayer membrane, which is made of a long carbon chain, and increase the aggregation number, thereby increasing the solubilization degree of the coronavirus phospholipid bilayer membrane. The applicant has prepared a cleaning composition utilizing this effect.

[0016] In the patent, when a complex phospholipid bilayer structure and micelles are formed using a substance having a branched structure, the packing density in the micelles can be reduced due to the repulsive force with the hydrophobic hydrocarbon, forming a structure that is much more favorable for dissolving the phospholipid bilayer. 13 H 27 (OCH2CH2) n OH is a substance synthesized from petrochemicals, and because it has many branched structures, it can be used for many purposes. However, it is not biodegradable and is not environmentally friendly.

[0017] In addition, the applicant discloses in Korean Patent Application No. 2020-0137612 a cleaning composition containing, based on a total of 100% by weight, 10 to 30% by weight of octylphenol ethoxylate (the number (n) of ethylene oxide contained in the ethoxylate is 2 to 15), 0.1 to 5% by weight of 2-ethylhexyl alcohol ethoxylate, 0.1 to 5% by weight of an ethylene oxide-propylene oxide block copolymer, and the balance water.

[0018] In the above patent, octylphenol ethoxylate is included as the main component responsible for removing and suppressing coronaviruses, and the octylphenol group in the octylphenol ethoxylate is a C8 alkyl group containing an aromatic phenol group, which binds well to the hydrophobic part consisting of a long carbon chain contained in the phospholipid membrane of coronaviruses, and the ethylene oxide part plays a role in imparting hydrophilicity to the hydrophobicity of the long carbon chain. At the same time, n, which represents the number of ethylene oxides, is adjusted to 2 to 15, preferably 5 to 10, and two or more substances with different numbers of n are mixed to adjust the hydrophilicity to a predetermined level for use.

[0019] However, although the main component, octylphenol ethoxylate, is effective in destroying the phospholipid bilayer membrane, it has a fatal drawback in terms of the environment because the alkyl group has a branched structure and the phenol component remains even after decomposition.

[0020] In addition to the above patents, Korean Patent Publication No. 2006-0079388 discloses prior art that has bactericidal and antibacterial properties against viruses, including SARS and influenza viruses, bacteria and fungi, and discloses a technology for using an inorganic antibacterial agent solution and its diluted substance in the form of a solution, such as a solution, prepared by preparing a nano-sized silver particle colloid and mixing it with one or more solvents selected from the group consisting of water, including distilled water, alcohols and surfactants within a specified concentration range, to synthesize a nano-sized titanium dioxide photocatalyst.

[0021] Furthermore, in Korean Patent No. 10-1317318, a 70-80% ethanol mixed solvent extract (content ratio = 4.3:1:7.2:16.3) consisting of gallic acid (1), a compound of para-digallic acid and meta-digallic acid (2), ethyl gallate (4), and a compound of ethyl para-digallate and ethyl meta-digallate (5) is obtained by a manufacturing method comprising the steps of cutting dried gallnut into small pieces, extracting the gallnut with a 70-80% ethanol mixed solvent of 10-20 times the dry weight at an extraction temperature of 20-100°C for 2-5 hours, repeatedly extracting the gallnut 1-5 times by cold immersion extraction, hot water extraction, ultrasonic extraction, or reflux cooling extraction, and then concentrating the extract under reduced pressure. The 70-80% ethanol mixed solvent extract (content ratio = 4.3:1:7.2:16.3) consisting of gallic acid (1), a compound of para-digallic acid and meta-digallic acid (2), ethyl gallate (4), and a compound of ethyl para-digallate and ethyl meta-digallate (5) is also obtained by a manufacturing method comprising the steps of: The present invention discloses an antibacterial composition for preventing and treating seasonal influenza or swine flu caused by an avian influenza virus selected from A / H1N1 seasonal influenza virus, A / H9N2 (A / Chicken / Korea / MS96 / 1996), or A / H1N1, comprising ethyl m-digallate and p-digallate as active ingredients.

[0022] With current technology, the development of products and vaccines to eliminate various viruses is still a long way off, and there is an urgent need to develop environmentally friendly disease prevention products that can effectively kill various viruses that have a phospholipid bilayer membrane envelope while not causing irritation when used in humans or animals. Summary of the Invention [Problem to be solved by the invention]

[0023] The object of the present invention is to provide an environmentally friendly epidemic prevention composition that has the function of killing phospholipid enveloped viruses, thereby effectively preventing viral transmission and infection by destroying the phospholipid membrane contained in enveloped viruses. [Means for solving the problem]

[0024] In order to achieve the above-mentioned object, the environmentally friendly epidemic prevention composition of the present invention, which has the function of killing phospholipid enveloped viruses, is characterized by containing, based on a total of 100% by weight, 10 to 40% by weight of polyoxyethylene lauryl ether, 0.01 to 10% by weight of polyoxyethylene 2-ethylhexyl ether, 0.01 to 10% by weight of polyoxyethylene polyoxypropylene alkyl ether, and the balance being a solvent.

[0025] According to one embodiment of the present invention, the SARS-CoV-2 virus removal rate in a dilution solution containing the composition in an amount of 0.1% by weight or less is characterized by being 100% in 1 minute.

[0026] Furthermore, according to one embodiment of the present invention, highly pathogenic avian influenza viruses in a dilute solution containing the composition in an amount of 0.1% by weight or less can be killed 100% within 1 to 10 minutes.

[0027] In addition, the composition of the present invention exhibits IC agonist activity against human epithelial keratinocyte cells (HaCaT), human bronchial epithelial cells (BEAS-2B), and human monocyte cells (THP-1). 50 It is characterized by having low cytotoxicity with a value (%) in the range of 0.003 to 0.0045.

[0028] Furthermore, the composition according to the present invention may have antibacterial activity against non-pathogenic and pathogenic strains.

[0029] The composition according to the present invention preferably maintains a neutral pH of 6 to 8.

[0030] The composition can be used by diluting it with water to a content of 0.01% to 0.05%.

[0031] According to one embodiment of the present invention, the remaining amount of water may further contain ethanol in an amount of 20% by weight or less.

[0032] According to one embodiment of the present invention, the composition may further contain a preservative, if necessary, in an amount of up to 0.1 wt % based on the total weight. Effect of the Invention

[0033] The environmentally friendly epidemic prevention composition of the present invention has a SARS-CoV-2 virus removal rate of 100% in 1 minute when diluted with an amount of 0.1% by weight or less, and is effective in killing 100% of highly pathogenic avian influenza viruses in 1 to 10 minutes.

[0034] In addition, IC50 activity against human epithelial keratinocyte cells (HaCaT), human bronchial epithelial cells (BEAS-2B), and human monocyte cells (THP-1) was also evaluated. 50 The value (%) is in the range of 0.003 to 0.0045, indicating that the compound has low cytotoxicity and antibacterial activity against both non-pathogenic and pathogenic strains. [Brief description of the drawings]

[0035] [Figure 1] FIG. 1 shows the measurement results of the SARS-CoV-2 virus removal rate of each composition according to Example 2 (the present invention), Comparative Example 1 (composition of Patent Document 1), and Comparative Example 2 (composition of Patent Document 2). [Diagram 2] FIG. 2 shows the results of measuring the killing effect of the composition of Example 2 (the present invention, Neutra Best L) against highly pathogenic avian influenza viruses. [Diagram 3]FIG. 3 shows the results of measuring the killing effect of the composition of Example 2 (the present invention, Neutra Best L) against highly pathogenic avian influenza viruses. [Figure 4] FIG. 4 shows the results of measuring the antibacterial activity of each composition for the same sample as in Experimental Example 3. [Diagram 5] FIG. 5 shows the results of measuring the antibacterial activity of each composition for the same sample as in Experimental Example 3. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] The present invention will now be described in more detail.

[0037] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0038] As used herein, the singular forms include the plural forms unless the context clearly dictates otherwise. Also, as used herein, "comprise" or "comprising" specifies the presence of stated features, numbers, steps, operations, members, elements and / or combinations thereof, but does not exclude the presence or addition of one or more other features, numbers, steps, operations, members, elements and / or combinations thereof.

[0039] The present invention provides an environmentally friendly antimicrobial composition capable of killing a variety of phospholipid enveloped viruses.

[0040] The composition according to the present invention has a composition containing, based on a total of 100% by weight, 10 to 40% by weight of polyoxyethylene lauryl ether, 0.01 to 10% by weight of polyoxyethylene 2-ethylhexyl ether, 0.01 to 10% by weight of polyoxyethylene polyoxypropylene alkyl ether, and the balance being a solvent.

[0041] The composition of the present invention preferably contains polyoxyethylene lauryl ether consisting only of a linear structure as a main component (meaning the component contained in the largest amount in the composition excluding the solvent). The polyoxyethylene lauryl ether is a nonionic surfactant that has excellent compatibility with anionic and cationic surfactants and exhibits excellent surface activity even in an aqueous solution in which acids, alkalis, salts, etc. coexist.

[0042] In recent experiments, the applicant has confirmed that the polyoxyethylene lauryl ether, which is composed only of a linear structure, can be used as the main component to more effectively remove the envelope of enveloped viruses compared to the case where the main component contains other components having conventional branched structures (Prior Patents 1 and 2).

[0043] This is because the alkyl groups of the phospholipids that compose the phospholipid bilayer membrane contained in the enveloped virus are linear. Therefore, in the present invention, by using a structure having a similar linear structure as the main component, it is expected that the affinity with the phospholipid bilayer membrane will be increased and the penetrating power when penetrating the phospholipid bilayer membrane will be improved.

[0044] In addition, after the phospholipid bilayer membrane is effectively removed, the polyoxyethylene lauryl ether needs to remain stable in an emulsion state in water. Even in this case, the degree of stability can be improved, and the polyoxyethylene lauryl ether, which is composed only of a linear structure, is biodegradable after use, making it possible to produce an environmentally friendly composition.

[0045] The polyoxyethylene lauryl ether is preferably contained in an amount of 10 to 40% by weight based on the total weight of the composition. If it is less than 10% by weight, the effect of removing the phospholipid bilayer membrane is insufficient, and if it exceeds 40% by weight, the content of other components becomes relatively small, which may be disadvantageous in preparing an environmentally friendly antiviral composition.

[0046] The composition of the present invention may contain polyoxyethylene 2-ethylhexyl ether in an amount of 0.01 to 10% by weight based on the total weight of the composition, so as to act as a penetration aid that enhances the penetration ability when penetrating between the phospholipid membranes of enveloped viruses. The polyoxyethylene 2-ethylhexyl ether has a low regularity structure with hydrocarbons substituted at the side chains, and therefore can create spacing when penetrating between the phospholipid bilayer membranes, which is expected to enhance the penetration ability.

[0047] Such polyoxyethylene 2-ethylhexyl ether is contained in an amount of 0.01 to 10% by weight based on the total weight of the composition. If it is less than 0.01% by weight, the desired effect cannot be obtained, and if it exceeds 10% by weight, the effect of further increasing the penetration power cannot be obtained, which is not preferable.

[0048] The composition of the present invention also contains polyoxyethylene polyoxypropylene alkyl ether produced by copolymerizing ethylene oxide and propylene oxide in an amount of 0.01 to 10% by weight based on the total weight of the composition.

[0049] The polyoxyethylene polyoxypropylene alkyl ether is added to suppress the generation of bubbles that may occur during the preparation of the environmentally friendly epidemic prevention composition having the function of killing phospholipid enveloped viruses according to the present invention, and is effective in removing bubbles when included in an amount of 0.01 to 10 wt % based on the total weight of the composition.

[0050] The cleaning composition according to the present invention is characterized by having a neutral pH of about pH 6 to 8. This has the advantage of minimizing damage or deterioration of the substance to which the composition of the present invention is applied, and is characterized by exhibiting an excellent virus killing effect.

[0051] The composition of the present invention further comprises a solvent in an amount remaining after removing the above-mentioned components. The solvent may be water or an organic solvent, and may be, but is not limited to, ordinary tap water or purified water from which all impurities contained in ordinary water, such as dissolved ions, solid particles, microorganisms, and organic matter, have been removed.

[0052] Furthermore, if necessary, an alcohol-based solvent such as ethanol or isopropyl alcohol may be contained as an organic solvent in an amount of up to 20% by weight based on the total solvent, which can reduce the viscosity of the final composition, remove air bubbles that may be generated during the production of the composition, and improve the storage properties of the composition when used at low temperatures, such as in winter.

[0053] In order to prevent problems such as mold growth during storage of the composition of the present invention and to improve storage stability, a known preservative may be contained in an amount of up to 0.1% by weight based on the total weight of the composition. The type of preservative is not particularly limited.

[0054] The eco-friendly epidemic prevention composition having the function of killing phospholipid enveloped viruses according to the present invention can be easily prepared by thoroughly mixing the above-mentioned components at room temperature (rt). The prepared composition has a hydrophobic-hydrophilic balance (HLB) value in the range of 12 to 15, and therefore, when used for various purposes as an eco-friendly epidemic prevention composition, it can form appropriate micelles in water to enhance the cleaning effect.

[0055] In particular, the composition according to the present invention is effective in killing various viruses having a phospholipid bilayer envelope, such as COVID-19, avian influenza, SARS (Severe Acute Respiratory Syndrome), MERS (Middle East Respiratory Syndrome), COVID-19 virus, and African swine fever.

[0056] In addition, the composition according to the present invention uses polyoxyethylene lauryl ether having a linear structure as a main component, and therefore can be effectively biodegraded after use, and can solve the problem of residual toxic organic substances in conventional surfactants having a branched structure, making it possible to prepare an environmentally friendly composition. Therefore, the composition according to the present invention can be safely used without any side effects when used for spraying on the human body, disinfecting hands, or for various animals.

[0057] The eco-friendly epidemic prevention composition prepared according to the present invention can be diluted with water and used for various desired purposes such as spraying, disinfection, cleaning, etc.

[0058] The dilution concentration can be 0.01% to 0.05% based on 100%, but is not limited to this, and it will be obvious to those skilled in the art that the dilution concentration can be adjusted depending on the intended use.

[0059] Preferred examples of the present invention will be described in detail below. However, the following examples are proposed for illustrative purposes only, and the scope of the present invention should not be construed as being particularly limited thereto. Furthermore, although specific compounds are exemplified in the following examples, it is clear to those skilled in the art that equivalents thereof can be used to achieve the same or similar effects. EXAMPLES

[0060] Examples and Comparative Examples: Preparation of Compositions According to the composition in Table 1 below, each composition was added to purified water, mixed at 18°C, and thoroughly stirred to prepare each epidemic prevention composition. In Table 1 below, Comparative Example 1 is the composition of Example 2 in Patent Document 1 of the applicant, and Comparative Example 2 is the composition of Example 2 in Patent Document 2. Each composition contained a component having a branched structure as a main component and was compared with the present invention.

[0061] [Table 1]

[0062] Experimental example 1: Measurement of SARS-CoV-2 antiviral effect The SARS-CoV-2 virus removal rate was measured for each of the compositions according to Example 2 (the present invention, Neutra Best L), Comparative Example 1 (composition of Patent Document 1, Neutra Best), and Comparative Example 2 (composition of Patent Document 2, Clean V) as follows. The results are shown in Figure 1.

[0063] Vero cells (monkey kidney epithelial cells) were added to a 96-well cell plate at 1 × 10 per well. 4 Then, 1 × 10 3 ~5×10 3 After dispensing each infectious virus into an EP tube, virus-cell culture medium (Gibco DMEM-0% FBS) was added so that the composition had a concentration of 0.1%, and the reaction was carried out for 30 seconds, 1 minute, 5 minutes, and 10 minutes.

[0064] After the reaction was completed, the cell culture medium was quickly frozen with dry ice, and the samples were stored in a deep freezer at -80°C. The prepared samples were diluted 10-fold in a 96-well U-bottom plate using cell culture medium. The cells cultured in the 96-well cell plate were washed once with 1x phosphate-buffered saline (PBS), and the serially diluted samples were transferred to the washed cells and reacted for 1 hour and 30 minutes. After the reaction was completed, the cell culture medium that had been in the plate was removed and replaced with a cell culture medium containing an appropriate concentration of TPCK trypsin.

[0065] The cell deformation and SARS-CoV-2 virus proliferation were visually observed under a microscope for 2 to 3 days, and surviving cells were stained with 10% crystal violet stain 3 days after infection. The results are shown in Table 2 and Figure 1 below.

[0066] [Table 2]

[0067] Table 2 above shows the results of measuring cell viability, and each composition according to Comparative Example 2 (composition of Patent Document 2, Clean V) showed a cell viability of 50% in 5 minutes and 25% in 10 minutes at a dilution concentration of 0.0375%. Comparative Example 1 (composition of Patent Document 1, Neutra Best) showed a cell viability in general at a dilution concentration of 0.0211%.

[0068] Next, referring to the results shown in FIG. 1, cells killed by viruses or cells killed by detergent toxicity were observed as transparent without staining, while surviving cells were stained with crystal violet and observed as dark purple. It was confirmed that Neutra Best L according to Example 1 of the present invention, at a dilution concentration of 0.0158%, showed 100% cell survival rate with all viruses killed in 1 minute and 10 minutes, and 75% survival rate in 5 minutes (see FIG. 1). In other words, it was confirmed that the composition according to the present invention removes 100% of the SARS-CoV-2 virus in a very short time of 1 minute.

[0069] Furthermore, at a dilution concentration of 0.0089% or less, all cells were killed by the virus, and the result was ND (Not Detected). These results show that the composition according to the present invention can kill all viruses at a concentration of 0.0158% or less.

[0070] These results confirm that Neutra Best L according to the present invention can effectively kill viruses while allowing cells to survive even at low concentrations.

[0071] Experimental Example 2: Measurement of the killing effect and cytotoxicity of highly pathogenic avian influenza (AI) viruses The highly pathogenic avian influenza virus killing effect and cytotoxicity (same as in Experimental Example 1) of the composition according to Example 2 (the present invention, Neutra Best L) were measured as follows, and the results are shown in Figures 2 and 3.

[0072] To investigate the effectiveness of killing highly pathogenic avian influenza viruses, a method called hemagglutinin assay using red blood cells (RBC) was used. Specifically, the principle that hemagglutinin, a protein in influenza viruses, causes hemolysis in RBCs is utilized. If the influenza virus is alive, hemolysis occurs using RBCs, causing the RBCs to burst and appear red overall. If the virus dies or the cells that grow the virus die, the virus is no longer present and a red dot is formed. This is because the plate used in this experiment has a U-bottom, and the RBCs gather at the bottom due to gravity, forming a dot.

[0073] Next, referring to Figure 2, which shows the killing effect of highly pathogenic avian influenza viruses, it can be seen that when the sample was diluted to a concentration of 0.0375%, red blood cells survived well, and all gathered as red dots in 1 minute, 5 minutes, and 10 minutes. On the other hand, at a concentration of 0.0281%, 50% of red blood cells survived in 1 minute, 25% in 5 minutes, and 75% in 10 minutes. As a result, it can be confirmed that at a concentration of 0.0375% or more, 100% of AI viruses can be killed in the extremely short time of 1 minute.

[0074] Referring to Figure 3 showing the results of the cytotoxicity experiment, it can be seen that at a dilution concentration of 0.0500%, regardless of the presence or absence of infection (○, ×), all cells are killed by the composition of the present invention and turn white. In contrast, at a dilution concentration of 0.0375% or less, it can be seen that all cells survive and turn purple. From these results, it was confirmed that there is no cytotoxicity at a concentration of 0.0375% or less.

[0075] Experimental Example 3: Measurement of cytotoxicity The toxicity of Example 2 (the present invention, NB-3), Comparative Example 1 (composition of Patent Document 1, NB-1), Comparative Example 2 (composition of Patent Document 2, NB-2), and the compositions shown in Table 3 below using commercially available products and various surfactants, against three types of cells, human epithelial keratinocyte cells (HaCaT), human bronchial epithelial cells (BEAS-2B), and human monocyte cells (THP-1), was evaluated as follows.

[0076] [Table 3]

[0077] 1. HaCaT, BEAS-2B, and THP-1 cell lines were seeded in a 96-well cell culture plate at 0.5 × 10 4 Cells / well, 0.5 x 10 4 Cells / well, 3 x 10 4 The cells were seeded at 100 cells / well. The reason for the difference in cell number is that the OD (optical density 570 nm) of the vehicle (Veh, the group treated with only the drug solvent) was set to about 1.0 when the cell number conditions were initially set.

[0078] 2. Treatment The cells were spread on a 96-well cell culture plate and treated with the drug after 24 hours. The amount of the drug to be dripped was diluted to about 1 / 100 of the medium volume (1 μl of the drug was dripped into 100 μl of medium). For example, in a 96-well plate, 100 μl of medium was used when spreading the cells. Therefore, if you want to treat a drug with a final concentration of 0.0075%, you need to prepare a drug with an initial concentration of 0.75%. In other words, if you drip 1 μl of a drug with a concentration of 0.75% into 100 μl of medium, the concentration that actually acts on the cells will be 0.0075%.

[0079] 3.MTT solution 24 hours after the cells were treated with a drug in advance, an MTT solution (1 mg / ml) was added in an amount equal to the volume of the medium, and the cells were allowed to react in an incubator for 2 to 3 hours. Here, the amount equal to the volume of the medium means that the medium is 100 μl and 100 μl of the MTT solution is added to it.

[0080] 4. Insoluble formazan When the reaction is complete, a blue-purple insoluble substance is formed on the bottom of the 96-well cell culture plate. This is a phenomenon that occurs when MTT tetrazolium is reduced to water-insoluble MTT formazan by mitochondrial cellular respiration.

[0081] Therefore, after the reaction, the supernatant was completely removed, and the water-insoluble MTT formazan was dissolved in DMSO, an organic solvent, and the OD was measured using a microplate reader. 570 The color intensity was quantified in nm.

[0082] IC indicates biological inhibitory concentration 50 A regression graph was drawn for each concentration, and the trend line was used to obtain the equation y=ax+b, from which the concentration of x for y=50 was calculated. The results are shown in Table 4 below.

[0083] [Table 4]

[0084] Referring to the results in Table 4 above, the IC 50 The value (%) is a confirmed value of cytotoxicity, and the lower the value, the higher the cytotoxicity. In the case of NB-3 according to the present invention, it was confirmed that it is an ingredient contained in the disinfectant for humidifiers, and has lower cytotoxicity than the commercial product (P2) that uses a cationic surfactant. In other words, it can be said that it has low cytotoxicity because it does not significantly affect normal human cells.

[0085] Experimental Example 4: Antibacterial activity The following experiment was carried out to evaluate the antibacterial activity of each composition against the same sample as in Experimental Example 3. The paper disc method was used for the antibacterial activity experiment. The strains and media used are shown in Table 5 below.

[0086] [Table 5]

[0087] Each strain was shake-cultured for 24 hours, and the resulting bacterial liquid was used as a seed culture. After preparing a medium suitable for each strain, 1.5% agar (Becton and Dickinson, BD, NJ, USA) was added and sterilized, the sterilized medium was cooled, and 1% of the seed culture liquid was added at about 40°C. Then, 20 ml of each was dispensed to prepare antibacterial active plate media. Each sample was absorbed into 8 mm (Advantec Ltd, Tokyo, Japan) at concentrations of 1, 10, and 100 μg / 50 μl, lightly placed on the surface of the antibacterial active medium, and cultured at 37°C for 18 to 24 hours. Then, the presence or absence of a growth inhibition zone (clear zone) was confirmed, and the diameter was measured. The results are shown in Tables 6 to 7 and Figures 4 to 5 below.

[0088] [Table 6]

[0089] [Table 7]

[0090] Referring to the results in Tables 6 to 7 and Figures 4 to 5, the composition (NB-3) according to the present invention had a minimum inhibitory concentration (MIC) value of 100 or more for non-pathogenic gram-positive strains such as Bacillus subtilis and gram-negative strains, and for pathogenic strains such as Salmonella typhimurium, and was 50 or more for the other strains.

[0091] In the case of pathogenic strains, the lower the MIC concentration, the better the antibacterial activity, but in the case of non-pathogenic strains, the absence of antibacterial activity is generally good. The composition of the present invention showed results consistent with this effect, although with some differences. [Industrial Applicability]

[0092] The antiviral composition of the present invention uses a substance having a linear structure as a main component, thereby increasing the affinity for the phospholipid membrane contained in enveloped viruses, improving the penetration power when penetrating the phospholipid bilayer membrane, and effectively dissolving the viruses, thereby making it possible to effectively remove various enveloped viruses.

[0093] In addition, after the phospholipid bilayer of an enveloped virus is destroyed, it needs to remain stable in an emulsion state in water, and even in this case, the degree of stability can be improved by the linear structure of the substance contained as the main component.

[0094] Furthermore, since the composition is likely to be biodegraded after use, it is possible to provide an epidemic prevention composition that is more environmentally friendly than the conventional compositions that use substances having a branched structure.

[0095] Therefore, the environmentally friendly epidemic prevention composition according to the present invention can be effectively used to eliminate enveloped viruses including COVID-19, avian influenza, SARS, African swine fever, etc.

Claims

1. Based on a total of 100% by weight, 10 to 40% by weight of polyoxyethylene lauryl ether; 0.01 to 10% by weight of polyoxyethylene 2-ethylhexyl ether; 0.01 to 10% by weight of polyoxyethylene polyoxypropylene alkyl ether; and a remaining amount of solvent, said composition being environmentally friendly and capable of killing phospholipid enveloped viruses.

2. The composition of claim 1, wherein the SARS-CoV-2 virus removal rate in a dilute solution containing the composition in an amount of 0.1% by weight or less is 100% in 1 minute.

3. The composition according to claim 1, which kills 100% of highly pathogenic avian influenza viruses in a dilute solution containing the composition in an amount of 0.1% by weight or less within 1 to 10 minutes.

4. The composition exhibited IC 100 activity against human epithelial keratinocyte cells (HaCaT), human bronchial epithelial cells (BEAS-2B), and human monocyte cells (THP-1). ５０ The composition according to claim 1, which has low cytotoxicity with a value (%) in the range of 0.003 to 0.0045.

5. The composition of claim 1 , wherein the composition has antibacterial activity against non-pathogenic and pathogenic strains of bacteria.

6. The composition of claim 1, wherein the composition maintains a pH of 6-8.

7. The composition according to claim 1, wherein the composition is used by diluting it with water at a content of 0.01% to 0.05%.

8. The composition of claim 1 , further comprising ethanol in an amount of up to 20% by weight of the remaining solvent.

9. The composition of claim 1 , further comprising a preservative in an amount of up to 0.1 wt % based on the total weight of the composition.

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