An environmentally friendly disease control composition that has the function 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 disease control composition having the function of killing phospholipid-enveloped viruses, and more specifically, to an environmentally friendly disease control composition that can effectively eliminate various phospholipid-enveloped viruses such as coronavirus, avian influenza, African swine fever, and severe acute respiratory syndrome. [Background technology]

[0002] Viruses have a very simple structure consisting of genetic material such as RNA or DNA and a protein shell (capsid) that surrounds it. Some viruses have a lipid membrane in addition to the protein shell, and those with this lipid layer (lipid layer, lipid membrane) are called 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 thought to play a major role in introducing their own genes into the host's genetic material. Inhibiting this mechanism is expected to allow for effective treatment of various types of viruses while preventing unwanted side effects.

[0004] Alcohol-based cleaning agents, which are currently widely used as cleaning agents to remove such viruses, have ethanol as their main component, which exerts its disinfecting effect by dissolving the virus's envelope (phospholipid), and are currently widely used in South Korea.

[0005] Ethanol dissolves the envelope of enveloped viruses, which are composed of a phospholipid layer and proteins, and either kills non-enveloped viruses outside the host or prevents them from penetrating cells and multiplying even if they manage to enter the host.

[0006] On the other hand, influenza viruses infect the respiratory system, causing systemic symptoms, and not only do they periodically change their form, but they also move to another host before the host dies without killing it. Scientists speculate that influenza viruses will survive until humanity is extinct. Influenza viruses are the viruses that cause the greatest economic loss to humankind, and although preventive vaccines have been developed, they have not been able to keep up with the virus's mutations, and a fundamental cure has yet to be found.

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

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

[0009] Outbreaks of avian influenza viruses, most likely to cause a fifth pandemic, are being reported continuously. Avian influenza A / H5N1, first discovered in 2003, originated in wild birds and poultry and spread to humans, killing 454 people in 16 countries by September 2017. In contrast, avian influenza A / H7N9, which emerged in China in 2013, has caused 599 deaths as of 2017.

[0010] On the other hand, SARS (Severe Acute Respiratory Syndrome) is a novel infectious disease that began in China in the winter of 2002 and spread throughout the world, including Hong Kong, Singapore, and Canada, within months. The causative pathogen is the SARS-associated coronavirus (SARS-coV). SARS-coV is known to be transmitted across species, from animals to humans, through a variant of an animal host coronavirus.

[0011] The primary routes of SARS transmission are known to be direct and indirect contact with mucous membranes via respiratory droplets or contaminated media. Symptoms include fever, fatigue, muscle pain, headache, and chills, and transmission is reported to primarily occur in the second week.

[0012] Patients suspected or believed to have SARS are isolated and hospitalized, and strict management is required to prevent transmission. However, since a vaccine or preventive medicine has not yet been developed, measures to prevent infection through direct contact are limited to avoiding travel to high-risk areas and thoroughly washing hands.

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

[0014] In Korean Patent Application No. 2020-0137611, the applicant provided an antiviral composition for preventing the spread of viruses, based on a total of 100% by weight, C 13 H 27 (OCH2CH2) n A detergent composition is disclosed comprising 10-30% by weight of OH (where n=2-15), 0.1-5% by weight of 2-ethylhexyl alcohol ethoxylate, 0.1-5% by weight of ethylene oxide-propylene oxide block copolymer, and the remainder being water.

[0015] C used as a main component in the patent 13 H 27 (OCH2CH2) n OH has a branched structure rather than a linear structure with 13 carbon chains. Therefore, it can bind well with the hydrophobic part consisting of long carbon chains contained in the phospholipid bilayer of the coronavirus, increase the aggregation number, and enhance the solubility of the phospholipid membrane of the coronavirus. The applicant has prepared a detergent composition by utilizing this effect.

[0016] In the patent, when forming a complex phospholipid bilayer structure and micelles using a substance with a branched structure, the packing density in the micelles can be lowered due to the repulsive force with hydrophobic hydrocarbons. Therefore, a structure much more favorable for the dissolution of the phospholipid bilayer can be formed. However, C 13 H 27 (OCH2CH2) n OH is a substance synthesized from petrochemistry. Since it has many branched structures, it can be applied to many uses, but it has the problem of non-biodegradability and is not environmentally preferable.

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

[0018] In the aforementioned patent, octylphenol ethoxylate is included as the main component responsible for the removal and suppression of coronavirus. The octylphenol group in the octylphenol ethoxylate is a C8 alkyl group containing an aromatic phenol group, which binds well to the hydrophobic portion consisting of a long carbon chain contained in the phospholipid membrane of the coronavirus, while the ethylene oxide portion plays a role in imparting hydrophilicity to the hydrophobicity of the long carbon chain. At the same time, the number 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 before use.

[0019] However, although octylphenol ethoxylate, which is included as the main component, has the effect of destroying the phospholipid bilayer, its alkyl group has a branched structure, and even after decomposition, the phenol component remains, which is a fatal drawback in the environment.

[0020] In addition to the aforementioned patent, as prior art possessing bactericidal and antimicrobial activity against viruses including SARS virus and influenza virus, bacteria, and fungi, Korean Patent Publication No. 2006-0079388 discloses a method for synthesizing an inorganic antimicrobial agent solution and its dilution in the form of a solution, vial, spray, aerosol, etc., produced by manufacturing nano-sized silver particle colloids and mixing them with one or more solvents selected from the group consisting of water including distilled water, alcohols, and surfactants within a predetermined concentration range, and using the resulting solution as a raw material to synthesize a nano-sized titanium dioxide photocatalyst.

[0021] Furthermore, in Korean Patent No. 10-1317318, dried Chinese gallnuts are cut into small pieces and extracted with a 70-80% ethanol mixed solvent that is 10-20 times the dry weight at an extraction temperature of 20-100°C for 2-5 hours, and extracted 1-5 times repeatedly by cold soaking extraction, hot water extraction, ultrasonic extraction or reflux cooling extraction, followed by concentration under reduced pressure. A 70-80% ethanol mixed solvent extract (content ratio = 4.3:1:7.2:16.3) consisting of gallic acid (1), a complex of para-digallic acid and meta-digallic acid (2), ethyl gallate (4), and a complex of ethyl para-digallate and ethyl meta-digallate (5), or penta-O-galloyl glucose, ethyl gallate, or a complex of ethyl para-digallate and ethyl meta-digallate separated therefrom is used as an active ingredient to contain a seasonal influenza or novel influenza caused by A / H1N1 seasonal influenza virus; or an antibacterial composition for the prevention and treatment of avian influenza caused by an avian influenza virus selected from A / H9N2 (A / Chicken / Korea / MS96 / 1996) or A / H1N1 is disclosed.

[0022] In the current technology, the development of products and vaccines for removing various viruses is still far from complete. There is an urgent need to develop environmentally friendly epidemic prevention products that can effectively kill various viruses with an envelope of phospholipid bilayer without causing irritation even when used in humans and animals.

Summary of the Invention

Problems to be Solved by the Invention

[0023] An object of the present invention is to provide an environmentally friendly epidemic prevention composition having a function of killing viruses with a phospholipid envelope, which can effectively prevent virus transmission and infection by destroying the phospholipid membrane contained in the envelope-bearing virus. [Means for solving the problem]

[0024] To achieve the above objective, the present invention provides an environmentally friendly disease control composition having the function of killing phospholipid-enveloped viruses, characterized by comprising, 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 remaining amount of solvent.

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

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

[0027] Furthermore, the compositions of the present invention provide ICs for human epithelial keratinocyte cells (HaCaT), human bronchial epithelial cells (BEAS-2B), and human monocyte cells (THP-1). 50 It is characterized by low cytotoxicity, with a value (%) in the range of 0.003 to 0.0045.

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

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

[0030] Furthermore, the composition can be used after being diluted with water at a concentration of 0.01% to 0.05%.

[0031] According to one embodiment of the present invention, the remaining 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 in an amount of 0.1 wt% or less of the total weight, if necessary. [Effects of the Invention]

[0033] The environmentally friendly disease control composition according to the present invention has a SARS-CoV-2 virus removal rate of 100% in 1 minute in a dilution containing 0.1% by weight or less, and is effective in killing 100% of highly pathogenic avian influenza viruses in 1 to 10 minutes.

[0034] Furthermore, ICs for human epithelial keratinocyte cells (HaCaT), human bronchial epithelial cells (BEAS-2B), and human monocyte cells (THP-1) were also performed. 50 It exhibits low cytotoxicity with a value (%) in the range of 0.003 to 0.0045, and has antibacterial activity against both non-pathogenic and pathogenic bacterial strains. [Brief explanation of the drawing]

[0035] [Figure 1] Figure 1 shows the measurement results of the SARS-CoV-2 virus removal rate for 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). [Figure 2] Figure 2 shows the measurement results of the highly pathogenic avian influenza virus killing effect of the composition according to Example 2 (the present invention, Neutra Best L). [Figure 3]Figure 3 shows the measurement results of the highly pathogenic avian influenza virus killing effect of the composition according to Example 2 (the present invention, Neutra Best L). [Figure 4] Figure 4 shows the measurement results of the antibacterial activity of each composition on the same sample as in Experimental Example 3. [Figure 5] Figure 5 shows the measurement results of the antibacterial activity of each composition on the same sample as in Experimental Example 3. [Modes for carrying out the invention]

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

[0037] The terms used herein are for the purpose of describing specific embodiments and are not intended to limit the invention.

[0038] As used herein, the singular form includes the plural form unless the context clearly indicates otherwise. Furthermore, as used herein, "comprise" or "comprising" identifies the presence of the described shape, number, stage, operation, component, element and / or combination thereof, and does not exclude the presence or addition of one or more other shapes, number, stage, operation, component, element and / or combination thereof.

[0039] This invention provides an environmentally friendly disease control composition that has the function of killing various phospholipid-enveloped viruses.

[0040] The composition according to the present invention comprises, 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 remaining amount of solvent.

[0041] The composition of the present invention preferably contains, as its main component (meaning the component present in the largest quantity in the composition excluding the solvent), a polyoxyethylene lauryl ether consisting solely of a linear structure. The polyoxyethylene lauryl ether is a nonionic surfactant that exhibits excellent compatibility with anionic and cationic surfactants and displays excellent surfactant activity even in aqueous solutions where acids, alkalis, salts, etc., are present.

[0042] In recent experiments, the applicant has confirmed that using the polyoxyethylene lauryl ether, which consists solely of a linear structure, as the main component allows for more effective removal of the envelope of enveloped viruses compared to cases where other components with a branched structure are used as the main component (Prior Patents 1 and 2).

[0043] This is because the alkyl groups of the phospholipids constituting the phospholipid bilayer contained in enveloped viruses are linear. Therefore, in this invention, by using a structure having a similar linear configuration as the main component, it was expected that the affinity with the phospholipid bilayer would be increased and the permeability when penetrating the phospholipid bilayer would be improved.

[0044] Furthermore, after effectively removing the phospholipid bilayer, the material needs to exist stably in an emulsion state in water. Stability can be improved even at this stage, and since the polyoxyethylene lauryl ether, which consists only of a linear structure, is biodegradable after use, it becomes possible to produce an environmentally friendly composition.

[0045] Such polyoxyethylene lauryl ether is preferably included in an amount of 10 to 40% by weight of the total weight of the composition. If the amount is less than 10% by weight, the effect of removing the phospholipid bilayer is insufficient, and if it exceeds 40% by weight, the content of other components will be relatively low, which may be detrimental to the preparation of 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 of the total weight of the composition, so as to act as a penetration aid to enhance penetration when it enters between the phospholipid membranes of enveloped viruses. Since the polyoxyethylene 2-ethylhexyl ether has a less regular structure in which hydrocarbons are substituted in the side chains, it can create gaps when it enters between the phospholipid bilayers, and is expected to enhance penetration.

[0047] Such polyoxyethylene 2-ethylhexyl ether is included in an amount of 0.01 to 10% by weight relative to the total weight of the composition. If the amount is less than 0.01% by weight, the desired effect will not be obtained, and if it exceeds 10% by weight, the effect of further enhancing the penetration power will not be obtained, which is undesirable.

[0048] Furthermore, the composition of the present invention contains polyoxyethylene polyoxypropylene alkyl ether, which is produced by copolymerizing ethylene oxide and propylene oxide, in an amount of 0.01 to 10% by weight relative to the total weight of the composition.

[0049] The aforementioned polyoxyethylene polyoxypropylene alkyl ether is added to suppress the generation of bubbles that may occur during the manufacture of the environmentally friendly disease control composition having the function of killing phospholipid enveloped viruses according to the present invention, and it is effective in removing bubbles when included in an amount of 0.01 to 10% by weight of the total weight of the composition.

[0050] Furthermore, the cleaning agent composition according to the present invention is characterized by having a neutral pH of approximately 6 to 8. This has the advantage of minimizing damage or alteration of the substance in which the composition of the present invention is used, and also exhibits excellent virus-killing effects.

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

[0052] Furthermore, if necessary, by including an alcohol-based solvent such as ethanol or isopropyl alcohol as an organic solvent in an amount of 20% by weight or less relative to the total solvent, the viscosity of the final composition can be reduced, bubbles generated during the production of the composition can be removed, and the storage properties of the composition when used at low temperatures such as in winter can be improved.

[0053] Furthermore, in order to prevent problems such as mold growth during storage of the composition according to the present invention and to improve storage stability, a known preservative may be included in an amount of 0.1% by weight or less relative to the total weight of the composition. The type of preservative is not particularly limited.

[0054] The environmentally friendly disease control composition according to the present invention, which has the function of killing phospholipid-enveloped viruses, 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 when used as an environmentally friendly disease control composition for various applications, it can form appropriate micelles in water to enhance the cleaning effect.

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

[0056] Furthermore, by using a linear polyoxyethylene lauryl ether as the main component, the composition according to the present invention is effectively biodegradable after use, solving the problem of residual toxic organic substances in conventional surfactants with branched structures, and enabling the preparation of an environmentally friendly composition. Therefore, the composition according to the present invention can be used safely without other side effects when used for applications such as spraying on the human body or disinfecting hands, or when applied to various animals.

[0057] The environmentally friendly disease control composition prepared according to the present invention can be diluted with water and used for various desired applications such as spraying, disinfection, and cleaning.

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

[0059] Preferred embodiments of the present invention will be described in detail below. However, the following embodiments are proposed for illustrative purposes only and should be interpreted as not particularly limiting the scope of the present invention. Furthermore, while specific compounds are illustrated in the following embodiments, it will be apparent to those skilled in the art that equivalent or similar effects can be achieved using their equivalents. [Examples]

[0060] Examples and Comparative Examples: Preparation of Compositions Each composition was prepared by adding the compositions shown in Table 1 below to purified water, mixing at 18°C, and stirring thoroughly. In Table 1 below, Comparative Example 1 is the composition of Example 2 in Patent Document 1 of the present applicant, and Comparative Example 2 is the composition of Example 2 in Patent Document 2. Compositions containing a branched structure as the main component were prepared and 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 composition 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] In a 96-well cell plate, place 1 x 10⁶ Vero cells (monkey kidney epithelial cells) per well. 4 Individual cultures were then performed. Next, 1 × 10 3 ~5×10 3 After dispensing individual infectious viruses into EP tubes, a viral cell culture medium (Gibco DMEM-0%FBS) was mixed in to a concentration of 0.1% of the composition, and the mixture was reacted for reaction times of 30 seconds, 1 minute, 5 minutes, and 10 minutes.

[0064] After the reaction was complete, the cell culture medium was rapidly frozen with dry ice, and the samples were stored in a deep freezer at -80°C. The prepared samples were diluted 10-fold in 96-well U-bottom plates using the cell culture medium. The cells cultured in the 96-well cell plates 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 complete, the cell culture medium that was previously in the plates was removed and replaced with cell culture medium containing TPCK trypsin at an appropriate concentration.

[0065] For 2-3 days, the deformation of cells and the proliferation of the SARS-CoV-2 virus were visually observed under a microscope, and three days after infection, surviving cells were stained with 10% crystal violet stain. 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. Each composition in Comparative Example 2 (composition of Patent Document 2, Clean V) showed a cell viability of 50% at a dilution concentration of 0.0375% and 25% at a dilution concentration of 10 minutes. Comparative Example 1 (composition of Patent Document 1, Neutra Best) showed generally good cell viability at a dilution concentration of 0.0211%.

[0068] Next, referring to the results shown in Figure 1, cells killed by the virus or by the toxicity of the cleaning agent were observed as clear and unstained, while surviving cells were stained with crystal violet and observed as a deep purple color. It was confirmed that Neutra Best L according to Example 1 of the present invention, at a dilution concentration of 0.0158%, killed all viruses at 1 minute and 10 minutes, showing a 100% cell viability rate, and showed a 75% viability rate at 5 minutes (see Figure 1). In other words, it was confirmed that the composition according to the present invention can remove 100% of the SARS-CoV-2 virus in a very short time of 1 minute.

[0069] Furthermore, at dilution concentrations of 0.0089% or less, all cells were killed by the virus and measured as ND (Not Detected). These results indicate that the composition according to the present invention can kill all viruses at concentrations 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) virus. 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. The results are shown in Figures 2-3.

[0072] To assess the effectiveness of killing highly pathogenic avian influenza viruses, a method called the hemagglutinin assay was used, which utilizes red blood cells (RBCs). Specifically, it employs the principle that RBCs undergo hemolysis due to a protein called hemagglutinin produced by the influenza virus. If the influenza virus is alive, hemolysis occurs using the RBCs, causing them to rupture and the entire surface to appear red. If the virus is killed, or if the cells necessary for viral replication are killed, the absence of the virus results in the formation of red spots. This is because the plate used in this experiment has a U-shaped bottom, causing the RBCs to gather at the bottom due to gravity and form a single point.

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

[0074] Referring to Figure 3, which shows the results of cytotoxicity experiments, it can be confirmed that at a concentration of 0.0500%, all cells were killed by the composition of the present invention, regardless of whether they were infected or not (○, ×), and the cells turned white. In contrast, at dilution concentrations of 0.0375% or less, all cells survived and the cells turned purple. From these results, it was confirmed that there is no cytotoxicity at concentrations of 0.0375% or less.

[0075] Experimental Example 3: Measurement of Cytotoxicity The toxicity of each composition shown in Table 3 below, using 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 a commercially available product and various surfactants, to 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. Place the HaCaT, BEAS-2B, and THP-1 cell lines into 0.5 × 10⁶ wells of a 96-well cell culture plate for cell seeding. 4 Cells / well, 0.5 × 10 4 Cells / well, 3 × 10 4 Cells were seeded per well. The difference in cell count is due to setting the OD (optical density at 570 nm) of the vehicle (Veh, the group treated only with the drug solvent) to approximately 1.0 when initially setting the cell count conditions.

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

[0079] 3.MTT solution After pre-treating the cells with a drug, add MTT solution (1 mg / ml) in an equal volume to the culture medium 24 hours later and incubate in an incubator for 2-3 hours. Here, "equal volume to the culture medium" means adding 100 μl of MTT solution to 100 μl of culture medium.

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

[0081] Therefore, after the reaction, the supernatant is completely removed, and the water-insoluble MTT formazan is dissolved in the organic solvent DMSO at the bottom, and the plate is analyzed using a microplate reader. 570 The intensity of the color was quantified using nm (nm).

[0082] IC, which indicates the concentration of biological inhibition 50 The values ​​(%) were calculated by plotting regression graphs for each concentration, obtaining the equation y=ax+b using trend lines, and calculating the concentration of x relative to y=50. The results are shown in Table 4 below.

[0083] [Table 4]

[0084] Referring to the results in Table 4 above, the IC25 concentration, which is the biological inhibitory concentration, is shown. 50 The % value indicates the level of cytotoxicity, with lower values ​​indicating higher cytotoxicity. In the case of NB-3 according to the present invention, it is an ingredient contained in humidifier disinfectants, and it was confirmed to have lower cytotoxicity compared to commercially available products (P2) using cationic surfactants. In other words, it does not have a significant effect on normal human cells, so it can be said to have low cytotoxicity.

[0085] Experimental Example 4: Antimicrobial Activity To evaluate the antibacterial activity of each composition against the same sample as in Experimental Example 3, the following experiment was conducted. The paper disc method was used for the antibacterial activity experiment. The bacterial strains and culture media used are shown in Table 5 below.

[0086] [Table 5]

[0087] Each bacterial strain was cultured with shaking for 24 hours, and the resulting bacterial suspension was used as the seed culture medium. After preparing a culture 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 medium was added at approximately 40°C. Subsequently, 20 ml portions were dispensed to prepare antimicrobial active plate media. Each sample was absorbed at concentrations of 1, 10, and 100 μg / 50 μl onto 8 mm (Advantec Ltd, Tokyo, Japan) plates, lightly placed on the surface of the antimicrobial active medium, and cultured at 37°C for 18-24 hours. After that, the presence or absence of growth inhibition zones (clear zones) was checked, and their diameter was measured. The results are shown in Tables 6-7 and Figures 4-5 below.

[0088] [Table 6]

[0089] [Table 7]

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

[0091] In the case of pathogenic bacterial strains, a lower MIC concentration indicates better antibacterial activity, but in the case of non-pathogenic bacterial strains, it is generally desirable to have no antibacterial activity. The compositions according to the present invention showed results consistent with this effect, although there were some differences. [Industrial applicability]

[0092] The antiviral composition according to the present invention, by using a substance having a linear structure as its main component, enhances its affinity for the phospholipid membrane contained in enveloped viruses, improves its penetration power when penetrating the phospholipid bilayer, and effectively dissolves various enveloped viruses, thereby effectively removing them.

[0093] Furthermore, after disrupting the phospholipid bilayer of the enveloped virus, it needs to exist stably in an emulsion state in water. In this state, the linear structure of the main component can improve its stabilization.

[0094] Furthermore, because it is highly likely to be biodegradable after use, it can provide an environmentally friendly disease control composition compared to using conventional materials with branched structures.

[0095] Therefore, the environmentally friendly disease control composition according to the present invention can be effectively used to eliminate enveloped viruses, including coronavirus, avian influenza, SARS, and African swine fever virus.

Claims

1. Based on a total of 100% by weight, 10-40% 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, An environmentally friendly disease control composition containing water as a solvent, which has the function of killing phospholipid-enveloped viruses.

2. The composition according to claim 1, wherein the SARS-CoV-2 virus removal rate in a dilution 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 dilution containing the composition in an amount of 0.1% by weight or less within 1 to 10 minutes.

4. The composition is used to induce IC (inducible conversion) in cells of human epidermal keratinocytes (HaCaT), human airway epithelial cells (BEAS-2B), and human monocyte cells (THP-1). ５０ The composition according to claim 1, wherein the value (%) has low cytotoxicity in the range of 0.003 to 0.0045.

5. The composition according to claim 1, wherein the composition has antibacterial activity against non-pathogenic and pathogenic bacterial strains.

6. The composition according to claim 1, wherein the composition has a pH of 6 to 8.

7. The composition according to claim 1, wherein the composition is used after being diluted in water at a concentration of 0.01% to 0.05%.

8. The composition according to claim 1, further comprising a preservative in an amount of 0.1 wt% or less of the total weight.

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