KR102305665B1 - Antiviral or antibacterial compositions comprising Dendropanax morbiferus - Google Patents

Abstract

The present invention relates to an antiviral or antibacterial composition including ethanol, purified water, glycerin, and a Dendropanax morbiferus extract or a Dendropanax morbiferus resin. The present invention can prevent viruses and infectious diseases.

Description

Antiviral or antibacterial compositions comprising Dendropanax morbiferus}

The present invention relates to an antiviral or antibacterial composition comprising hwangchil, and more particularly, to an antiviral or antibacterial composition comprising ethanol, purified water, glycerin, and hwangchil tree extract or hwangchil tree resin.

Due to the 2020 novel coronavirus infection (COVID-19) epidemic, trials and sufferings not only in Korea but also around the world are ongoing. Meanwhile, scientists from 32 countries around the world sent an open letter to the World Health Organization (WHO), suggesting an airborne infection of the novel coronavirus infection (COVID-19), urging them to revise the precautions against COVID-19.

Researchers at MIT have published a study showing that an aerosol containing a virus can travel up to 8 meters. In addition, the global medical journal 'New England Journal of Medicine' published an analysis that the Corona 19 virus can float in the air for up to 3 hours in the form of microscopic particles of less than 5 microns (one millionth of a meter).

On the other hand, the WHO said that the corona 19 virus can be transmitted even in indoor places such as homes, schools, sports institutions, hospitals, and multi-use facilities, and the virus can remain in a closed and crowded room for a long time and infect it.

In the prior art related to antiviral agents, antiviral agents against coronavirus, including herbal extracts such as Chopi tree, Cucurbita mulberry tree, nurijang tree, arrowroot tree, spruce tree, ginseng, fig tree, black magnolia tree, spearmint tree, oleander tree and ephedra, etc. (Patent Document 1), an antiviral composition containing lauric acid as an active ingredient (Patent Document 2), maple syrup, tabasheer, uiiin, manuka honey and an antiviral agent for treatment or alleviation of adenovirus, including propolis ( Patent Document 3) and the like are disclosed.

On the other hand, as the prior art regarding hwangchil tree extract, hwangchil tree extract having anticancer activity (patent document 4), hwangchil extract having hepatocellular protective effect, hwangchil fraction and pharmaceutical composition containing them (patent document 5), antibacterial activity, etc. Although the seed extract of Hwangchil tree excellent in physiological activity (Patent Document 6) and the like are disclosed, the antiviral activity of the Hwangchil tree extract is not known.

Laid-open Patent Publication No. 10-2004-0097445 Registered Patent Publication No. 10-0998081 Unexamined Patent Publication No. 10-2020-0024186 Registered Patent Publication No. 10-0318019 Laid-Open Patent Publication No. 10-2003-0079205 Laid-open Patent Publication No. 10-2006-0131360

The present invention has been devised in order to solve the problems of the prior art as described above, and an object of the present invention is to provide an antiviral or antibacterial composition having not only antiviral activity but also antibacterial and antifungal activity by including a hwangchil tree extract will do

The present invention provides an antiviral or antibacterial composition comprising (A) ethanol, purified water, glycerin and (B) hwangchil tree extract or hwangchil tree resin in order to achieve the above object.

Antiviral or antibacterial composition comprising hwangchil according to the present invention can prevent viruses and infectious diseases by sterilizing and disinfecting pathogens such as bacteria and fungi as well as viruses floating in the air with fine particles.

Figure 1a shows the state of E. coli at the initial stage of inoculation.
Figure 1b shows the state of E. coli after 5 minutes of culture.
Figure 2a shows the state of pneumococci at the initial stage of inoculation.
Figure 2b shows the state of pneumococcus after 5 minutes of culture.
Figure 3a shows the state of the MRSA bacteria in the initial inoculation.
Figure 3b shows the state of MRSA bacteria after 5 minutes of culture.
Figure 4a shows the state of Staphylococcus aureus at the initial stage of inoculation.
Figure 4b shows the state of Staphylococcus aureus after 5 minutes of culture.
Figure 5a shows the state of Candida at the initial stage of inoculation.
Figure 5b shows the state of Candida after 5 minutes of culture.
Figure 6a shows the state of the streptococci at the initial stage of inoculation.
Figure 6b shows the state of streptococci after 5 minutes of culture.
Figure 7a shows the state of Pseudomonas aeruginosa at the initial stage of inoculation.
Figure 7b shows the state of Pseudomonas aeruginosa after 5 minutes of culture.
Figure 8a shows the state of Bacillus at the initial stage of inoculation.
Figure 8b shows the state of Bacillus bacteria after 5 minutes of culture.
Figure 9a shows the body weight change of 300 mg / kg BW (stage 1) rats administered.
Figure 9b shows the change in body weight of 300 mg/kg BW (step 2) rats administered.
Figure 9c shows the body weight change of the dose of 2000 mg / kg BW (stage 3) rats.
Figure 9d shows the change in body weight of mice administered dose 2000 mg / kg BW (stage 4).

The present invention relates to an antiviral or antibacterial composition comprising (A) ethanol, purified water, glycerin, and (B) hwangchil tree extract or hwangchil tree resin.

The antiviral or antibacterial composition of the present invention is based on 100 parts by weight of the ethanol, 30 to 45 parts by weight of the purified water, 1 to 5 parts by weight of the glycerin, and 0.1 to 20 parts by weight of the hwangchil tree extract or hwangchil tree resin may include

The antiviral or antibacterial composition of the present invention is preferably based on 100 parts by weight of the ethanol, 35-40 parts by weight of the purified water, 2-4 parts by weight of the glycerin, and 10 ~ 15 parts by weight.

The antiviral or antibacterial composition of the present invention is more preferably based on 100 parts by weight of ethanol, 38 to 39 parts by weight of the purified water, 2 to 2.5 parts by weight of the glycerin, and 12 to 12 to the hwangchil tree extract or hwangchil tree resin. 13 parts by weight.

The antiviral or antibacterial composition according to an embodiment of the present invention contains, based on the total weight, 65% by weight of ethanol, 25% of purified water, 1.5% by weight of glycerin, and 8% by weight of the hwangchil tree extract or hwangchil tree resin can, but is not limited thereto.

In the antiviral or antibacterial composition of the present invention, when it contains in excess of the content of ethanol, the fine particles of the ethanol are excessively scattered when the composition is sprayed in the air in the form of an aerosol, spray or mist. , it is necessary to be careful because it can cause viral infection or respiratory disease.

Hwangchil ( Dendropanax morbiferus Lev.) is an evergreen tree of the dicotyledonous umbel family, which can be widely used for wood, metal, glass, etc. It is said that a refreshing benzoic scent is emitted after applying it, and its pharmacological action is said to be effective for quadriplegia and menstrual irregularities.

The hwangchil tree extract used in the antiviral or antibacterial composition of the present invention is collected and dried leaves, stems, roots, or mixtures thereof, washed 1 to 5 times with distilled water and pulverized, and then the hwangchil leaves , a viscous extract obtained by adding a solvent 1 to 30 times the weight of the stem, root, or mixture thereof, heating the extracted extract, and concentrating the filtrate obtained above under reduced pressure, or obtained through the reduced pressure concentration It may be a powder obtained by drying the viscous extract with a freeze dryer, but is not limited thereto.

The solvent used in the preparation of the hwangchil tree extract may be water or an alcohol having 1 to 4 carbon atoms, and the alcohol having 1 to 4 carbon atoms may be methanol, ethanol, isopropanol, butanol, but is not limited thereto.

The hwangchil wood resin used in the antiviral or antibacterial composition of the present invention is a resin paint collected from the hwangchil tree and is also called hwangchil (黃漆). and has a bitter taste. The main component of the essential oil contained in the hwangchil is sesquiterpene, and other alcohols, esters, and the like are contained.

The antiviral or antibacterial composition of the present invention may further include 0.1 to 2 parts by weight, preferably 0.5 to 1 parts by weight, of (C) fragrance, based on 100 parts by weight of the ethanol, the content of which is less than 0.1 parts by weight This is because the predetermined effect is insignificant, and even if it exceeds 2 parts by weight, it is not economical because it does not exhibit any more functional effects.

The fragrance included in the antiviral or antibacterial composition of the present invention is peppermint oil, green peppermint oil, anise oil, allspice oil, cardamom oil, cassia oil, cinnamon oil, citrus oil, clove oil, coriander oil, eucalyptus oil, fennel oil, Nutmeg oil, leaf oil, lavender oil, lavandin oil, rock cherry oil, fruit flavoring agent, flower flavoring agent, limonene, cineol, menthol, linalol, anetol, eugenol, menthone, carbonone, methyl salicylate and cinnamaldehyde It may be selected from the group consisting of, but is not limited thereto.

The formulation of the antiviral or antibacterial composition of the present invention may be a solution, tablet, capsule, pill, powder, suspension or granule, or may be formulated in the form of an aerosol, spray, mist or droplet, but is limited thereto no.

The antiviral or antibacterial composition of the present invention may be, for example, sprayed from an air sterilizer, but is not limited thereto.

The antiviral or antibacterial composition of the present invention is methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus , Escherichia coli , Pseudomonas aeruginosa , Candida sp., strep It may exhibit antibacterial activity against at least one selected from Streptococcus sp., Klebsiella pneumoniae and Bacillus sp., but the antibacterial spectrum is not limited thereto.

The antiviral or antibacterial composition of the present invention may exhibit antiviral activity against one or more selected from influenza A (H1N1) virus, human rhinovirus A (HRV) and enterovirus 71 (EV71). The antiviral spectrum is not limited thereto.

Hereinafter, the present invention will be described in detail through examples. However, these examples are only for describing the present invention in detail, and the scope of the present invention is not limited by these examples.

<Preparation Example> Preparation of Hwangchil tree extract

Hwangchil leaves collected from the Hwangchil tree (Dendropanax morbiferus H. Lev.) that grow wild in the southern regions of Korea and Jeju Island were naturally dried in the shade. 12 kg of the dried hwangchil leaves were put into an extractor, 180 L of purified water (Duksan Pharmaceutical Co., Ltd.) was added, and low temperature extraction was performed for 24 hours.

The extract was filtered under reduced pressure with filter paper (Whatman No. 1, Whatman, Maidstone, UK), and the filtrate was finally filtered to obtain a hwangchil tree extract having a concentration of 8%. The final extract after all the processes is around 120L and refrigerated after filling.

<Example> Preparation of antiviral or antibacterial composition

Ethanol (Palmco Ethanol), purified water (Duksan Pharmaceutical Co., Ltd.), glycerin (Duksan Pharmaceutical Co., Ltd.), Hwangchil tree extract (Hwangchil extract) or Hwangchil resin and fragrance (limonene) of the above preparation are mixed. An antiviral or antibacterial composition was prepared, and the mixing ratio is shown in Table 1 (unit: parts by weight) below.

division Example comparative example One 2 3 4 5 6 One 2 ethanol 100 100 100 100 100 100 100 100 Purified water 30 37.5 45 30 37.5 45 37.5 45 glycerin One 2.5 5 One 2.5 5 2.5 5 Hwangchil Extract 5 12.5 20 - - - - - Hwangchil resin - - - 5 12.5 20 - - Spices One One One One One One One One

<Experimental Example 1> Antiviral activity test In order to measure the virus proliferation inhibitory ability of the composition prepared in Example 2, 2x10 ⁴ HeLa cells were placed in each well of a 96-well plate and cultured for 24 hours. After 24 hours, the culture supernatant from each well was removed, and the influenza A (H1N1) virus solution, the human rhinovirus A (HRV) solution and the enterovirus 71 (EV71) solution titrated with _{TCID 50 were added to each well.} Each concentration of the composition was administered to each well of a 96-well plate so that each concentration was 100, 10, 1, 0.1 ㎍ / ㎖.

After the infection test was completed, 100 μl of 10% trichloroacetic acid (TCA) was added to each well of a 96-well plate, and left at 4° C. for 1 hour, followed by washing with distilled water several times. After drying at room temperature, 100 μl of a 0.4% (w/v) sulforhodamine B (SRB) solution dissolved in 1% (v/v) acetic acid was added and stained for 30 minutes. SRB staining solution not bound to cells was washed several times with 1% (v/v) acetic acid and then dried again.

100 μl of a 10 mM Tris solution (pH 10.5) was added to each well to sufficiently dissolve the cell-bound dye, and then absorbance was measured at 560 nm.

Each treatment group was denoted as an untreated group (A), a group treated only with the composition (B), a group treated with only the virus (C), and a group treated with the virus and the composition (D), and 1 of the composition The cell viability (%) at a concentration of μg/ml and the antiproliferative ability (%) of the composition against viruses were calculated and shown in Table 2 below.

At this time, the cell viability was calculated according to Equation 1 below, and the proliferation inhibitory ability was calculated according to Equation 2 below.

division H1N1 HRV EV71 cell viability 175.8 179.1 182.0 proliferation inhibition 95.29 96.12 95.83

<Experimental Example 2> Antibacterial activity test

To test the antimicrobial activity of the composition prepared in Example 2, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 4352, MRSA ( Staphylococcus aureus) subsp. aureus ATCC 33591), Staphylococcus aureus (Staphylococcus aureus ATCC 6538), Candida (Candida albicans ATCC 10231), Streptococcus (Streptococcus mutans ATCC 25175), Pseudomonas aeruginosa (Pseudomonas aeruginosa ATCC 15442) and Bacillus bacteria (Bacillus cereus ATCC 11778 ), the results of the antibacterial test are shown in Tables 3 to 5 and FIGS. 1A to 8B below.

As shown in Tables 3 to 5 and FIGS. 1A to 8B, it can be seen that the antimicrobial activity of the composition according to the present invention is very excellent.

<Experimental Example 3> Deodorization test

For the deodorization test of the composition prepared in Example 2, ammonia, trimetalamine, hydrogen sulfide, and methylmercaptan were subjected to a deodorization test in accordance with KCL-EL608:2018, a domestic and international official test standard. 10 mL of ammonia, 5 mL of trimetalamine, 20 mL of hydrogen sulfide, and 20 mL of methyl mercaptan were added to the open space to measure the concentration reduction (%), and the test results are shown in Table 6 below.

<Experimental Example 4> Acute oral toxicity test

In order to evaluate the toxic reaction that appears during acute oral administration of the composition (hereinafter referred to as the test substance) prepared in Example 2, a test was performed according to the National Institute of Environmental Sciences Notice No. 2020-28. Using SD female rats, the test substance was administered at a dose of 300 mg/kg B.W. (Steps 1 and 2) and 2000 mg/kg B.W. (Steps 4 and 5) were divided into 3 animals each, and a total of 12 mice were orally administered once. Mortality, general symptoms, and weight changes were observed for 14 days after administration, and surviving animals were necropsied to visually check for organ abnormalities.

During the experimental period, no animals were killed due to the administration of the test substance. The results of observation of general symptoms are shown in Tables 7 and 8 below, and no abnormal findings were observed due to administration of the test substance. In addition, the weight measurement results are shown in Table 9 and FIGS. 9A to 9D, and no change in body weight due to administration of the test substance was observed in all administration groups. The autopsy findings are shown in Table 10 below, and no abnormal findings were observed in all administration groups.

<Experimental Example 5> Acute skin irritation and corrosion test

In order to evaluate the acute skin irritation and corrosion (Corrosive) of the composition (hereinafter referred to as the test substance) prepared in Example 2, a test was performed according to the National Institute of Environmental Sciences Notice No. 2019-23.

After 0.5 mL of the test substance was administered to the back of a rabbit (NZW rabbit) for 4 hours, mortality, general symptoms, weight change, and skin irritation and corrosion were evaluated for 72 hours.

During the experimental period, no animals were killed due to the administration of the test substance. As a result of observation of general symptoms, no specific abnormal symptoms were observed in all experimental animals.

As a result of body weight measurement, no weight loss was observed due to administration of the test substance.

As a result of observing the site of administration of the test substance, skin irritation and corrosion were not observed in the initial test, and corrosion was observed at 72 hours after removing the patch for 4 hours in the confirmatory test. The mean score of skin reaction for erythema and eschar formation was calculated as “0.0” in the initial test and “0.7” in one confirmation test (1201), and in one confirmation test (1202). Corrosion was observed at 72 hours, so it was not possible to calculate the average skin response score (mean score, the average score of irritation at 24, 48, and 72 hours). At 1, 24, 48, and 72 hours after removing the information on the test substance, the stimuli were scored according to Table 11 below, and the skin response scores are shown in Table 12 below.

<Experimental Example 6> Eye irritation and corrosion test

In order to evaluate the eye irritation and corrosion of the composition (hereinafter referred to as the test substance) prepared in Example 2, a test was performed according to the National Institute of Environmental Sciences Notice No. 2019-23.

After administering 0.1 mL of the test substance to the right eye (conjunctival sac) of a rabbit (NZW rabbit), weight change, mortality, general symptoms, and eye irritation and corrosion for 6 days were evaluated.

During the experimental period, no animals were killed due to the administration of the test substance. As a result of observation of general symptoms, lacrimation was observed in the initial test.

As a result of body weight measurement, no weight loss was observed due to administration of the test substance.

Regarding the observation and evaluation of the administration site, after administration of the test substance, eye mucosal reactions were observed up to 1, 24, 48, 72 hours and 5 days in the initial test and 1, 24, 48, 72 hours and 6 days in the confirmation test. As a result, conjunctivae redness and chemosis were observed. The ocular mucosal irritation observed after administration of the test substance was recovered within 5 days in the initial test and within 6 days in the confirmation test, and reversibility was confirmed. The mean ocular mucosal response score for conjunctival flare (Mean score, mean score of stimulation at 24, 48, 72 hours) was calculated as “1.7” in the initial test and “0.3” and “1.0” in the confirmation test. The mean score of the ocular mucosal reaction for conjunctival edema was calculated as “0.0” in the initial test and “0.7” and “0.7” in the confirmation test. At 1, 24, 48, and 72 hours after administration of the test substance, the stimuli were scored according to Table 13, and the mean ocular mucosal response scores are shown in Table 14 below.

Although described with reference to preferred embodiments of the present invention as described above, those of ordinary skill in the art can view the present invention within the scope without departing from the spirit and scope of the present invention as set forth in the following claims. It will be understood that various modifications and variations of the invention are possible.

Claims (8)

(A) ethanol, purified water, glycerin and
(B) In the antiviral composition comprising a hwangchil tree extract,
Based on 100 parts by weight of the ethanol, 35-40 parts by weight of the purified water, 2-3 parts by weight of the glycerin, and 10-15 parts by weight of the Hwangchil tree extract,
The composition is an antiviral composition, characterized in that it exhibits antiviral activity against one or more selected from influenza A (H1N1) virus, human rhinovirus A (HRV) and enterovirus 71 (EV71). delete According to claim 1, wherein the hwangchil tree extract is a solvent extract of hwangchil tree, the solvent is water or an antiviral composition, characterized in that the alcohol having 1 to 4 carbon atoms. According to claim 1, Based on 100 parts by weight of the ethanol, (C) The composition for antiviral, characterized in that it further comprises 0.1 to 2 parts by weight of fragrance. 5. The method of claim 4, wherein the fragrance is peppermint oil, green peppermint oil, anise oil, allspice oil, cardamom oil, cassia oil, cinnamon oil, citrus oil, clove oil, coriander oil, eucalyptus oil, fennel oil, nutmeg oil, leaf oil, lavender Oil, lavandin oil, rock cherry oil, fruit flavoring agent, flower flavoring agent, limonene, cineol, menthol, linalol, anetol, eugenol, menthone, carbonic acid, methyl salicylate, and cinnamaldehyde selected from the group consisting of Antiviral composition, characterized in that. The composition for antiviral according to claim 1, wherein the composition is formulated in the form of an aerosol, spray, mist or droplet. delete delete

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