KR102138654B1 - Antiviral composition comprising natural substance-derived substance and phytoncide - Google Patents

Abstract

The present invention relates to an antiviral composition comprising a phytoncide component extracted from cypress tree and a substance derived from a natural product, and a use thereof.

Description

Antiviral composition comprising natural substance-derived substance and phytoncide {ANTIVIRAL COMPOSITION COMPRISING NATURAL SUBSTANCE-DERIVED SUBSTANCE AND PHYTONCIDE}

The present invention relates to an antiviral composition comprising a phytoncide component extracted from a cypress and a substance derived from a natural product, and its use.

With the recent influenza (H1N1) and coronavirus (SARS, MERS, COVID-19, etc.) spread around the world, various analyzes and outlooks on the symptoms, spread rates and aftermath of the outbreak are pouring out, and countries around the world have established measures to prevent them. I am on the sidelines.

Therefore, in order to prevent virus infection, the concept of'space defense' has risen one step further from the concept of'contact defense' such as washing hands and wearing a mask, and the interest in products related to this has increased.

'Spatial defense' refers to a method of reducing the likelihood of virus transmission by inactivating or killing viruses in the space itself where viruses may be present, in order to minimize the possibility of infection of various bacteria or viruses floating in the air in a specific space. do. That is, it is a preventive method for minimizing the virus infection in the daily living space such as a house, office, elevator, restaurant, car, hotel, infant and childcare facilities, or public places such as churches, educational centers, schools, and concert halls.

In particular, according to the World Health Organization (WHO), the H1N1 and coronaviruses are reported to have a high likelihood of being infected by droplets that are transmitted from person to person and are mostly infected by droplets spreading in the air through coughing or sneezing. Therefore, the importance of the prevention of virus particles present in the atmosphere of the space where many people work or live is increasing day by day.

On the other hand, Cypress ( Chamaecyparis obtusa ) is also known as cypress, and is an evergreen tree of the oleaceae family of the outer plant, which is native to Japan, but has been widely cultivated as a planted species in the southern regions of Korea through development, due to its peculiar fragrance. It is used as a deodorant and antibacterial agent.

Phytoncide, which is produced from cypress trees, is a self-defense material for plants, and is a substance released or secreted by plants to resist pathogens, pests, and fungi, and contains sterilizing and insecticidal ingredients in itself. The constituents of phytoncide are organic compounds composed of phenolic compounds including terpenes, alkaloid components, and glycosides, and are known to have antibacterial, sedative, deodorizing, and stress-relieving effects.

The phytoncide is a natural substance, not a chemical synthetic substance, and is absorbed by the human body without difficulty, and is known to have numerous functions such as antibacterial action, deodorant action, sedative action, and stress relief action. In addition, phytoncide is known to cleanse and moisturize the skin with excellent sterilization, antibacterial, and cleaning effects, and to strengthen the immune function in the body.

As described above, various studies have been conducted on how to utilize phytoncide's antibacterial activity, air purification ability, stress relief, strengthening intestinal and cardiopulmonary functions, and indoor sterilization, but spraying it in the air can show virus suppression effect. There have not been enough studies yet, and there is a problem in that the preference is lowered when a high concentration of phytoncide is used to exhibit sufficient antibacterial and antiviral effects.

Korean Patent Publication No. 10-2017-0027536

An object of the present invention is to provide a composite composition of cypress water containing an extract derived from natural products having antiviral efficacy and phytoncide and a method for manufacturing the same.

Another object of the present invention is to provide a method for inhibiting or inactivating viruses, including various application forms and uses of the composition.

One aspect of the present invention for solving the above problems is 1200 to 1800 parts by weight of cypress water obtained by hot water extraction of leaves and stems of cypress; And myrrh (Commiphora myrrha Holmes), a directed tree (Boswellia Carterii), honeysuckle (Lonicera japonica), row (杏仁), DEG (桃仁), octagonal (八角, Illicium verum Hook. F .), Bongchul (蓬朮, Curcuma zedoaria ), a mixed extract of one or more selected from the group consisting of Hibiscus syriacus , Poncirus trifoliata seeds, and Phellinus igniarus ; 1 to 100 parts by weight; containing, provides an antiviral composition.

Specifically, the cypress water is obtained by adding distilled water to a cypress leaf and a cypress tree trunk, extracting hot water at 100° C. for 2 to 3 hours, and then concentrating under reduced pressure, and the mixed extract is 90° C. using 60 to 70% ethanol. It may be obtained by extraction with hot water for 3 to 4 hours.

Specifically, the mixed extract is 30 parts by weight of Myrrha Holmes ( Commiphora myrrha Holmes ), 20 parts by weight of Boswellia Carterii , 15 parts by weight of Lonicera japonica , 10 parts by weight of Hangin, 10 parts by weight of Doin Part, 10 parts by weight of the Illicium verum Hook. f. , 15 parts by weight of curcuma zedoaria, 10 parts by weight of Hibiscus syriacus , 5 parts by weight of seeds of Poncirus trifoliata and horsefly Mud mushroom ( Phellinus igniarus ) may be prepared by mixing 5 parts by weight and extracting it with hot water at 90° C. for 3 to 4 hours with 60 to 70% ethanol, but is not limited thereto.

In the present invention, specifically, the extract derived from the natural product may be extracted with water, C ₁ to C ₄ lower alcohol, or a mixed solvent thereof. More specifically, it may be extracted using distilled water or 60 to 70% ethanol, but is not limited thereto.

Specifically, the extraction process of the extract may be used alone or in combination with a method selected from one or more of the group consisting of cold, warm, reflux, and ultrasonic extraction, and is not limited thereto, and appropriately changed and applied as necessary. Can.

In addition, after performing the extraction or fractionation process, the extract may be concentrated under reduced pressure filtration or additionally concentrated and/or lyophilized to remove the solvent or specifically concentrated under reduced pressure after extraction. After the extract can be obtained, it is not limited thereto, and can be suitably changed and applied.

In the present invention, the “phytoncide” is a 100% natural material that is difficult to artificially synthesize or combine, consisting of phenolic compounds, terpene, alkaloid components, and glycoside components. In general, it is rapidly absorbed into the human body through a forest bath and is human. It is a substance that has various effects such as strong antibacterial and antifungal action, stress relief, skin soothing effect, sound sleep inducing effect, allergic skin disease improvement and immune function enhancement by selectively sterilizing harmful bacteria.

As a result, for busy modern people who cannot frequently take a forest bath, phytoncide ingredients are added to various products that can come into contact with everyday life, such as fragrances, face washes, cosmetics, and medicines. Losing.

As a result of research on phytoncide, not only the amount of phytoncide emitted from plants, but also its efficacy are all different, and a large amount of phytoncide occurs in conifers such as pine or pine trees, especially in cypress trees (扁栢, Chamaecyparis obtusa ). It is known to generate phytoncide, which has the highest content of ingredients and strong healing power.

Currently, products containing phytoncide ingredients mainly use essential oils or cypress water extracted from cypress trees, where essential oils are flowers, buds, leaves, stems, roots, and natural resins of plants ( It is a volatile substance with a strong scent obtained from 天然樹脂) and contains phytoncide.

In the present invention, the "antiviral" means inactivation of the pathogen virus, preferably permanent inactivation.

Specifically, the virus capable of being inactivated or permanently inactivated by the composition may be a DNA virus and an RNA virus, and specific examples thereof are influenza A virus (human, bird, swine (new)), influenza B virus, parainfluenza Virus, hepatitis virus (A~E), measles virus, herpes virus, mumps virus, rabies virus, norovirus, HIV virus, SARS, avian influenza, swine flu, Ebola virus, MERS coronavirus (MERS-CoV) And COVID-19 may be one or more selected from the group consisting of, and most specifically, may be influenza virus A or herpes virus, but is not limited thereto.

In the experimental example of the present invention, the antiviral efficacy of the composition of the present invention was confirmed using the influenza virus A and herpes virus (Table 1).

Another aspect of the present invention relates to a cosmetic composition comprising the composition.

The cosmetic composition of the present invention is a solution, ointment for external use, cream, foam, nutrient lotion, soft lotion, pack, softener, emulsion, makeup base, essence, soap, liquid detergent, bathing agent, sunscreen cream, sun oil, suspension, emulsion Liquid, paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch and spray can be prepared in a formulation selected from the group consisting of, specifically, soap , It may be a liquid cleaning agent or hand cream, but is not limited thereto.

The cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers formulated in general skin cosmetics, for example, oil, water, surfactant, moisturizer, lower alcohol, thickener as a common ingredient , Chelating agents, pigments, preservatives, fragrances, etc. may be appropriately blended, but is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention may vary depending on the formulation of the cosmetic composition.

When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. as carrier components This may be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component, and in the case of a spray, additionally, chlorofluorohard Propellants such as locarbon, propane/butane or dimethyl ether, but are not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butyl glycol oil, and the like can be used, and in particular, cottonseed oil, peanut oil, corn seed oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan May be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, micro Crystalline cellulose, aluminum metahydroxide, bentonite, agar or trakant may be used, but is not limited thereto. These may be used alone or in combination of two or more.

In addition, the composition of the present invention can be used as a transdermal administration method such as applying or spraying directly on the skin, and the route of administration of the composition of the present invention can be administered through any general route as long as it can reach the target tissue.

The amount of the composition of the present invention can be appropriately adjusted according to the individual difference or formulation such as age, degree of lesions, etc., and can be used once to several times a day to the skin for 1 week to several months.

The antiviral composition comprising the natural product-derived material provided by the present invention and cypress water containing phytoncide has excellent virus suppression effect and excellent formulation safety.

In addition to phytoncide, the composition includes a number of extracts having antiviral efficacy derived from natural products, and thus can exhibit excellent virus suppression even at low concentrations, thereby solving problems of low preference such as causing discomfort caused by using high concentrations of phytoncide.

In addition, the composition of the present invention is derived from natural products and is not toxic, and even when applied to the human body, it does not cause side effects and thus has high utilization.

The antiviral composition of the present invention and the spray using the same have an effect of preventing damage (infection) caused by bacteria or viruses from a place such as a hospital, lecture room, public transportation, etc., which is a main route of infection in an early stage.

In addition, the anti-viral composition of the present invention and the spray using the same are quick to dry and sticky even though the above-described characteristics are continuously expressed due to the composition of the present invention remaining on the inner wall, outer wall, ceiling, article of a place such as a hospital, lecture room, public transportation, etc. There is no practical effect to use.

It should be understood that the effects of the present invention are not limited to the above-described effects, and include all effects that can be deduced from the configuration of the invention described in the detailed description or claims of the present invention.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are only to illustrate the present invention, the present invention is not limited by the following examples.

Example 1. Preparation of extracts derived from natural products having antiviral efficacy

Before preparing the antiviral composition of the present invention, an extract derived from natural products having antiviral efficacy was prepared.

First, in order to prepare the extract derived from the natural product, myrrh ( Commiphora myrrha Holmes ), frankincense tree ( Boswellia Carterii ), gold silver ( Lonicera japonica ), hangin (杏仁), doin (桃仁), octagon (八角, Illicium verum Hook. f. ), Bongchul (蓬朮, Curcuma zedoaria ), Hibiscus syriacus , Poncirus trifoliata seeds, and Phellinus igniarus are prepared by washing and removing water, using 60 to 70% ethanol Then, hot water was extracted at 90° C. for 3 to 4 hours.

Specifically, the extract derived from the natural product is 30 parts by weight of Commiphora myrrha Holmes , 20 parts by weight of Boswellia Carterii , 15 parts by weight of Lonicera japonica , 10 parts by weight of passersby, 10 parts by weight of Doin 10 parts by weight, 10 parts by weight of octagonal octagonal hook ( f. ), 15 parts by weight of curcuma zedoaria, 10 parts by weight of Hibiscus syriacus , 5 parts by weight of Poncirus trifoliata seeds, and 5 parts by weight of Phellinus igniarus was mixed and prepared by extracting hot water at 90°C for 3 to 4 hours with 60 to 70% ethanol.

Example 2. Preparation of antiviral composition containing phytoncide

Before preparing the antiviral composition containing phytoncide, cypress water was prepared for mixing with the extract derived from the natural product prepared in Example 1.

To prepare the cypress water, 0.5 kg of dried cypress leaf and 0.5 kg of cypress tree trunk are crushed and placed in an extraction container, and 15 L of distilled water is mixed with 1 kg of cypress leaf and stem to heat for 2 to 3 hours at 100°C. And extracted with hot water, filtered through a 400 mesh filter cloth, concentrated with a vacuum rotary concentrator, filtered again, and then extracted twice more in the same process using the same amount of distilled water, filtered and concentrated under reduced pressure to contain phytoncide. Cypress water was prepared.

The extract of Example 1 based on 1500 parts by weight of cypress water obtained through the above production method is 1 to 100 parts by weight, more specifically 10 to 50 parts by weight, and most specifically 30 parts by weight of the present invention Virus composition was prepared, but is not limited thereto.

Preparation of Comparative Examples 1 to 4

In the antiviral composition of Example 2, Comparative Examples 1 to 4 were prepared with different compositions as follows.

Comparative Example 1 was prepared in the same manner as in Example 2, except that frankincense tree was excluded when preparing the composition of Example 1, Comparative Example 2 further excluded gold and silver coins from Comparative Example 1, and compared Example 3 was prepared in the same manner as in Example 2, except that Example 1 was prepared by including 10 parts by weight of the composition of Example 2, Comparative Example 4 is 100 parts by weight of Example 1 in the composition of Example 2 It was prepared in the same manner as in Example 2, except that it was prepared.

Experimental Example 1. Confirmation of cytotoxicity and virus suppression effect

For the compositions of Example 2 and Comparative Examples 1 to 4, experiments were conducted to confirm cytotoxicity and virus inhibitory effects.

Specifically, for the experiment, the RNA virus influenza virus A (A/Brisbane/59/2007, NCCP 42464) and the DNA virus Herpes simplex virus (NCCP 43109) were distributed from Korea University Pathogenic Virus Bank (KBPV). The cells for culturing were used as vero cells (African green monkey kidney cell line), and prepared by maintenance passage using Dulbecco's modified Eagle's medium (DMEM) medium containing 5% FBS and penicillin/streptomycin.

Cytotoxicity of the host cells to the compositions of Examples 2 and 1 to 4 was measured by a cell attachment test. Specifically, after inoculating each well of a 96-well plate by suspending 4.0×10 ⁴ cells/100 μL of vero cells in a culture medium, 5% CO ₂ , incubated at 37° C. for 24 hours, 70-80% monolayer ( monolayer) was formed, washed once with PBS, and inoculated with 100 μL of the culture medium and 100 uL of the composition. Subsequently, the cell lesion effect was observed microscopically for 2 days to confirm whether the cells did not cause toxicity due to adhered proliferation.

The virus for inoculation was cultured in a tissue culture flask of 75 cm ² (Corning, USA), and when the attached cells were detached due to cell lesion effect, freezing and thawing were repeated twice, followed by centrifugation at 1,500×g for 5 minutes. Cell debris was removed and the supernatant containing the virus was subdivided in a cryogenic vial (Nalgene, USA) and stored at -80°C.

Infection virus titer TCID ₅₀ (50% tissue culture infective dose) was calculated, and as a result, it was confirmed that influenza virus A was 3.36×10 ² /mL and herpes virus was 3.26×10 ⁷ /mL.

The antiviral efficacy test for the extract was inoculated with 100 μL of each of the culture medium for inoculation containing the compositions of Examples 2 and Comparative Examples 1 to 4 and 10 times the virus of 10 times the amount of TCID ₅₀ , and after 2 days of culture, the cytotoxicity test was performed. Conversely, the degree of inhibition of the cell lesion effect of the composition was observed under a microscope. Normal cells per sample, cells inoculated with viruses only, and cells to which only the compositions of Examples 2 and 1 to 4 were added to the 96-well plate were set as controls for each of 2 wells.

As a result of the cytotoxicity test, it was confirmed that both of Example 2 and Comparative Examples 1 to 3 did not show cytotoxicity under the above conditions, except for Comparative Example 4 prepared by including 100 parts by weight of the composition of Example 1.

In addition, the results of experiments for confirming the virus suppression ability were divided into not able to inhibit the virus (-), partially inhibited (+), and completely inhibited (++), and the results are shown in Table 1 below.

division Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Restraint ++ + - + ++

As shown in Table 1, it was confirmed that the composition of Example 2 of the present invention completely inhibited the virus, and Comparative Example 1 excluding the frankincense tree had a slightly reduced virus suppression ability, and Comparative Example excluding the frankincense tree and gold silver coin 2 confirmed that the effect was significantly reduced compared to other experimental groups. Comparative Example 3 caused a decrease in effect by adding a small amount of the composition of Example 1, and Comparative Example 4 exhibited an excellent virus inhibitory effect, but since it was confirmed that the cytotoxicity test was the only one to show cytotoxicity, it was applied to the human body. It seems somewhat unreasonable to follow.

Experimental Example 2. Confirmation of user's sensory evaluation during spraying

As confirmed in Experimental Example 1, the composition of Example 2 is expected to be able to prevent a viral infection by applying an auto-injector or a humidifier to suppress the virus present in the air in a certain space using excellent virus suppression ability. A sensory evaluation was performed when the compositions of Example 2 and Comparative Example were sprayed in the air.

At this time, Comparative Example 4, which is likely to exhibit cytotoxicity, was excluded, and only Comparative Examples 1 to 3 were targeted.

The sensory test for the antiviral composition was carried out by selecting 24 adult males and females in their 20s and 50s. Specifically, 6 groups of each age and gender were evenly distributed, and 1 ml was sprayed in a closed 1 square space. Subjects' scent preferences, discomfort (strong odor), dizziness, annoyance, and overall preference were evaluated.

The 10-point rating method was used, and the higher the preference, the higher the score. When the evaluation was completed for each person, ventilation was completely performed while taking sufficient rest for 30 minutes, and all of them were repeatedly evaluated three times to obtain an average value. The results are shown in Table 2 below.

division Example 2 Comparative Example 1 Comparative Example 2 Comparative Example 3 Incense preference 9.1 8.3 7.7 8.2 Discomfort (strong smell) 9.0 7.4 7.2 8.5 Dizziness 8.9 8.3 8.1 8.5 Tortoise 8.7 8.0 7.8 8.2 Overall preference 8.9 7.9 7.5 8.1

As shown in the results shown in Table 2, it was confirmed that the frankincense tree extract and the gold silver coin extract had a large effect on the overall flavor preference, and showed significantly lower results compared to Example 2 in both discomfort and overall preference. Was confirmed. In addition, the results of Comparative Example 3 confirmed that when the composition of Example 1 is included in less than the numerical range suggested by the present invention, the preference decreases to the extent that human perception can be perceived in general.

Experimental Example 3. Inhalation toxicity test

Inhalation toxicity when the composition of Example 2 of the present invention was injected into the air was confirmed.

Specifically, for 5 mice (BALB/c) male and female, the composition of Example 2 was exposed to concentrations of 0, 125, 250, and 500 mg/m ³ for 4 hours each day, resulting in 14 days of mortality, general symptoms, and body weight. Changes and autopsy findings were observed.

The test criteria were conducted in accordance with the'Standards for Toxicity Testing of Drugs' and'Standards for Drug Safety Test Management' announced by the Food and Drug Administration.

The experimental results, because the abnormality is not observed in the highest concentration of 500 mg / m ³ in death rates, general symptoms, weight change, autopsy findings, even if the suction at a high concentration (500mg / m ³⁾ not noticeable toxicity was confirmed to be safe.

The above description of the present invention is for illustration only, and those skilled in the art to which the present invention pertains can understand that it can be easily modified to other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and equivalent concepts should be interpreted to be included in the scope of the present invention.

Claims (6)

Cypress trees 1200 to 1800 parts by weight obtained by hot water extraction of leaves and stems of cypress trees; And
Myrrh (Commiphora myrrha Holmes), a directed tree (Boswellia Carterii), honeysuckle (Lonicera japonica), row (杏仁), DEG (桃仁), octagonal (八角, Illicium verum Hook. F .), Bongchul (蓬朮, Curcuma zedoaria) , Mixed extract of Hibiscus syriacus , Poncirus trifoliata seeds, and Phellinus igniarus ; In 1 to 100 parts by weight; containing, in the composition for antiviral,
The virus is an influenza virus A or herpes virus, antiviral composition. According to claim 1,
The cypress water is obtained by adding distilled water to cypress leaves and cypress trunks, extracting hot water at 100° C. for 2-3 hours, and then concentrating under reduced pressure.
The mixed extract is obtained by extracting hot water for 3 to 4 hours at 90 ℃ using 60 to 70% ethanol, anti-viral composition. According to claim 2,
The mixed extract is 30 parts by weight of myrrha holmes ( Commiphora myrrha Holmes ), 20 parts by weight of frankincense tree ( Boswellia Carterii ), 15 parts by weight of gold silver ( Lonicera japonica ), 10 parts by weight of passersby, 10 parts by weight of doin, octagonal (八角, Illicium verum Hook.f.) 10 parts by weight, 15 parts by weight of curcuma zedoaria, 10 parts by weight of Hibiscus syriacus , 5 parts by weight of Poncirus trifoliata seeds and horsefly mud mushroom ( Phellinus igniarus ) 5 parts by weight of the mixture, it was prepared by extracting hot water for 3 to 4 hours at 90 ℃ with 60 to 70% ethanol, antiviral composition. delete A cosmetic composition comprising the antiviral composition of any one of claims 1 to 3. An antiviral water dispersion composition for spraying comprising the antiviral composition of any one of claims 1 to 3.