KR20220083218A - Composition for anti-coronavirus containing extract of Prunus genus - Google Patents

Abstract

The present invention is an anti-coronavirus composition comprising an extract of a plant of the genus Prunus as an active ingredient; And it relates to a pharmaceutical composition, food composition, health functional food, quasi-drug composition, cosmetic composition and feed additive for the prevention, improvement or treatment of coronavirus infection comprising the composition.
In the present invention Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata ), Prunus pendura ( Prunus pendula ), Prunus cerasus ( Prunus cerasus ), Prunus leveilleana ( Prunus leveilleana ) , Prunus sargentii ( Prunus sargentii ) or a mixture extract thereof is characterized in that it inhibits the entry of coronavirus into host cells by inhibiting TMPRSS2 and ACE2 gene expression, preventing, improving or treating coronavirus infection For pharmaceutical composition, food composition, health functional food, quasi-drug composition such as hand sanitizer, cosmetic composition or feed additive, etc. can be utilized.

Description

Composition for anti-coronavirus containing extract of Prunus genus plant {Composition for anti-coronavirus containing extract of Prunus genus}

The present invention is an anti-coronavirus composition comprising an extract of a plant of the genus Prunus as an active ingredient, a pharmaceutical composition, food composition, quasi-drug composition for prevention, improvement or treatment of coronavirus infection comprising the composition , to cosmetic compositions and feed additives.

Corona virus disease 19 (COVID-19), which is currently prevalent around the world, is a respiratory syndrome caused by infection with the novel coronavirus SARS-CoV-2, and SARS-CoV-2, which causes it, is It is an RNA virus belonging to Coronaviridae. So far, Corona Virus Infectious Disease-19 is known to be spread through droplets (saliva) or contact, and when infected, it is characterized by various respiratory infections ranging from mild to severe, such as fever, malaise, cough, dyspnea, and pneumonia. Although the level of fatality rate by country and age is very different, the global fatality rate reaches an average of about 3.85% (WHO, as of August 1, 2020), which poses a great threat to the world not only in health and medical care but also socially and economically. there is a situation

On the other hand, corona virus has a structure in which protrusion-shaped proteins (spike proteins) are tightly attached to the outer surface of the virus membrane, which binds strongly to the host cell and supports the virus to enter the host cell quickly. do. As a result of analyzing the three-dimensional structure of SARS-CoV-2 through cryo-EM analysis, it was confirmed that it was very similar to the SARS virus spike protein (Walls et al., 2020; Wrapp). et al., 2020), it was recently found that, due to this morphological similarity, SARS-CoV-2 is also strongly attached to the surface of host cells through the angiotensin-converting enzyme 2 (ACE2) receptor (Zhou et al., 2020, Yan et al., 2020). In addition, the virus penetrates into the cell only after the protein scissors present in the host cell cut off a part of the spike protein after binding. In the case of SARS-CoV-2, TMPRSS2 (transmembrane protease serine subtype 2) in the respiratory cell membrane is was found to be used as (Hoffmann et al., 2020).

As such, the combination of coronavirus and host cell corresponds to the first stage of viral infection, so researchers are using a strategy to block this stage before infection in the development of vaccines or initial therapeutics. However, the development of materials that are safe and effective for the human body that can be used to prevent SARS-CoV-2 infection is still insignificant, and it is expected that a considerable amount of time will be required in the future.

Under this background, the present inventors are derived from natural products and effectively inhibit TMPRSS2 and ACE2 gene expression without adversely affecting the human body to prevent corona virus infection in advance or to improve or treat infectious diseases caused by coronavirus As a result of diligent efforts to develop ( Prunus leveilleana ), Prunus sargentii ( Prunus sargentii ) or a mixed extract thereof has reached to complete the present invention by confirming that TMPRSS2 and ACE2 gene expression is significantly suppressed. The composition of the present invention including it may be utilized as a pharmaceutical composition or a food composition for the prevention, improvement or treatment of corona virus infection, including coronavirus disease 19 (corona virus disease 19).

SUMMARY OF THE INVENTION The present invention aims to solve the above problems and other problems related thereto.

An exemplary object of the present invention is to provide a composition for anti-coronavirus, comprising an extract of a plant of the genus Prunus as an active ingredient.

Another exemplary object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of coronavirus infection comprising the anti-coronavirus composition.

Another exemplary object of the present invention is a food composition for the prevention or improvement of coronavirus infection comprising the anti-coronavirus composition; And to provide a health functional food comprising the food composition.

Another exemplary object of the present invention is to provide a quasi-drug composition, cosmetic composition, and feed additive for the prevention or improvement of coronavirus infection, including the anti-coronavirus composition.

The technical problem to be achieved according to the technical idea of the invention disclosed in this specification is not limited to the problem for solving the above-mentioned problems, and another problem not mentioned can be clearly understood by those skilled in the art from the description below. There will be.

This will be described in detail as follows. Meanwhile, each description and embodiment disclosed in the present application may be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present application fall within the scope of the present application. In addition, it cannot be seen that the scope of the present application is limited by the detailed description described below.

One aspect of the present invention for achieving the above object provides a composition for anti-coronavirus, comprising an extract of a plant of the genus Prunus as an active ingredient.

In particular, the Prunus ( Prunus ) Extracts of plants of the genus Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata ( Prunus serrulata ), Prunus pendura ( Prunus pendula ), Prunus cerasus ( Prunus cerasus ) , Prunus leveilleana ( Prunus leveilleana ), Prunus sargentii ( Prunus sargentii ) or a mixed extract thereof, a composition comprising the same, by inhibiting TMPRSS2 and ACE2 gene expression, the corona virus entering the host cell It is characterized by suppressing Due to these characteristics, the present invention can be usefully used as a pharmaceutical composition or a food composition for preventing, improving or treating infection of a coronavirus, including coronavirus disease 19 (COVID-19).

In the present invention, the term "Prunus ( Prunus ) genus" refers to the only genus belonging to the Prunus genus, that is, the Prunus spp. Specifically, the Prunus genus plant in the present invention is Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata ), Prunus pendura ( Prunus pendula ), Prunus cerasus ( Prunus cerasus ), may mean, but is not limited to, Prunus leveilleana , or Prunus sargentii . "Prunus cerasoides" is distributed in the Himalayas from Himachal Pradesh in northern India to southwestern China, Burma and Thailand, and is mainly cultivated as an ornamental tree. "Prunus cerulata", also called Japanese cherry and East Asian cherry, is grown as an ornamental for cherry blossom festivals and exhibitions. "Prunus pendura" is also called All-Cherry tree because its flowers bloom before the leaves and at the same time it blooms earlier than other cherry trees, and it is distributed in Korea and Japan. When the branches hang down like a weeping willow, it is called a drooping cherry tree ( P. pendula ) and is planted in gardens and parks. "Prunus cerasus" is mainly distributed in Europe and southwest Asia, and is widely used in soups, cakes, tarts, and other dishes. "Prunus lebaileana" blooms in April and the fruit is a round drupe and ripens dark red in June. "Prunus sargenti" is a species distributed mainly in East Asia, also called wild cherry, has the advantage of less disease and a long lifespan, and the fruits ripening in May and June can be eaten by humans.

In the present invention, the term "extract" refers to an extract or stock solution obtained by extraction of a target substance, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a prepared or purified product of the extract, or a mixture thereof, etc. , including extracts of all formulations that can be formed using the extract itself and the extract. The method of extracting the extract of the present invention is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, cold extraction, solvent extraction, steam distillation, elution method, compression method, juice extraction, ultrasonic extraction, filtration, reflux extraction, etc. It can be carried out using a combination of methods. In addition, the type of extraction solvent used for extraction of the extract is not particularly limited, and any solvent known in the art may be used. Specifically, the extraction solvent may be one or more selected from the group consisting of water, an organic solvent including an alcohol having 1 to 4 carbon atoms, an inorganic solvent, and a mixed solvent thereof.

Specifically, the extract in the present invention is Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata ( Prunus serrulata ), Prunus pendura ( Prunus pendula ), Prunus cerasus ( Prunus cerasus ), Prunus leveile Ana ( Prunus leveilleana ), Prunus sargentii ( Prunus sargentii ) It may refer to an extract, or a mixed extract thereof. The plant of the genus Prunus may be purchased and used commercially sold, or harvested or grown in nature, but is not limited thereto. Specifically, the extract of a plant of the genus Prunus in the present invention may be a methanol extract or a juice extract of a plant of the genus Prunus, but is not limited thereto.

In addition, the extract of the present invention may be extracted from the stem, leaf, bark, flower, branch, fruit, root, heartwood, outpost, or a combination thereof of a plant of the genus Purnus, but is not limited thereto.

In the present invention, the term "anti-coronavirus" includes all of improving, preventing, or treating diseases mediated/infected by the corona virus, inhibiting the infection, entry and proliferation of cells in an individual of the corona virus. Specifically, the anti-coronavirus may prevent, improve, or treat a disease mediated/onset by the coronavirus by inhibiting the corona virus from entering the host cell, and a disease mediated/onset by the coronavirus For example, there may be, for example, coronavirus disease 19 (corona virus disease 19), but is not particularly limited thereto. The effect of the composition of the present invention inhibiting the entry of the corona virus into the host cell may be shown by inhibiting the expression of TMPRSS2 and/or ACE2 gene.

The "TMPRSS2 (transmembrane protease serine subtype 2)" is an enzyme protein belonging to a type 2 transmembrane serine protease, and when it binds to a type 2 transmembrane domain, a type A receptor domain capable of binding calcium, and other cell envelopes or molecules It is composed of group A scavenging receptors rich in cysteine used. This is implicated in several physiological and pathological processes, which have a role to play when the coronavirus enters the cell, and the proteases TMPRSS2 and TMPRSS11a, which are more frequently seen in the envelope of cells in the respiratory system, in particular, are the SARS coronaviruses. (SARS-CoV) helps to enter the cell. In addition, TMPRSS2 and TMPRSS11d are known to facilitate the invasion of viruses through coordination with the ACE2 receptor.

The "ACE2 (angiotensin-converting enzyme 2)" is an enzyme attached to cell membranes such as lung, artery, heart and kidney, and serves as an entry point into the cell of some coronaviruses including SARS-CoV. is known Specifically, the binding of the spike S1 protein of SARS-CoV and SARS-CoV-2 with the enzymatic domain of ACE2 present on the cell surface causes both viruses and enzymes present in the cell to move into the cell, and the endocytosis ( known to cause endocytosis.

In one embodiment of the present invention, it was confirmed that when several species of plant extracts of the genus Prunus, including the Prunus cerasoides extract, were treated, the transcriptional inhibitory activity of TMPRSS2 and ACE2 mRNA was remarkably excellent.

Another aspect of the present invention for achieving the above object provides a pharmaceutical composition for the prevention or treatment of coronavirus infection, comprising the anti-coronavirus composition comprising the anti-coronavirus composition. Specifically, the corona virus infection may refer to an infectious disease caused by a coronavirus, and specifically may refer to a coronavirus disease 19 (corona virus disease 19), but is not limited thereto.

The term "prevention" of the present invention refers to any action that inhibits or delays corona virus infection through administration of the pharmaceutical composition of the present invention, and the term "treatment" of the present invention refers to any act of inhibiting or delaying corona virus infection by administering the pharmaceutical composition of the present invention. It means any action in which an infection is improved, alleviated, or is beneficially altered.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent commonly used in the preparation of a pharmaceutical composition in addition to the extract of the plant of the genus Prunus. Specific examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil may be used, but is not limited thereto.

In addition, the pharmaceutical composition of the present invention can be prepared according to a conventional method for tablets, pills, powders, granules, capsules, suspensions, internal solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions and freeze-drying agents, respectively. It may have any one dosage form selected from the group consisting of, and the dosage form may be oral or parenteral in various forms. In the case of formulation, it may be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants, but is not limited thereto. Specifically, tablets, pills, powders, granules, capsules, etc. may be used in the solid preparation for oral administration, and the solid preparation may include at least one excipient, for example, starch, calcium carbonate, sucrose or lactose. ), gelatin, etc. may be used. In addition, lubricants such as magnesium stearate and talc may be used in addition to simple excipients, but the present invention is not limited thereto. In addition, as liquid formulations for oral administration, suspensions, internal solutions, emulsions, syrups, etc. may be used, and various excipients, for example, wetting agents, sweeteners, fragrances, in addition to commonly used simple diluents such as water and liquid paraffin, Preservatives and the like may be used. A sterile aqueous solution, a non-aqueous solution, a suspension, an emulsion, a freeze-dried formulation, or a suppository may be used as a formulation for parenteral administration. Examples of the non-aqueous solvent and suspending agent include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). This document is considered a part of this specification.

The optimal amount and dosing interval of individual administration of the pharmaceutical composition of the present invention will be determined by the nature and extent of the disease being treated, the dosage form, route and site, and the age and health condition of the individual, and the doctor will ultimately use the appropriate dosage to be used. will be determined by one of ordinary skill in the art. Such dosing may be repeated as often as appropriate. If side effects occur, the dosage and frequency may be changed or reduced according to normal clinical practice. In addition, the administration route of the composition of the present invention may be administered through any general route as long as it can achieve the object of the present invention. For example, the pharmaceutical composition of the present invention may be administered intraperitoneally, intravenously, subcutaneously, intradermally, orally, but is not limited thereto.

The pharmaceutical composition of the present invention may be administered to mammals such as mice, dogs, cats, cattle, horses, pigs, and humans through various routes.

Another aspect of the present invention for achieving the above object is a food composition for the prevention or improvement of coronavirus infection comprising an anti-coronavirus composition; And it provides a health functional food comprising the food composition.

Specifically, the corona virus infection may refer to an infectious disease caused by a coronavirus, and specifically may refer to a coronavirus disease 19 (corona virus disease 19), but is not limited thereto.

As used herein, the term "improvement" refers to any action in which the symptoms of coronavirus infection are improved or beneficially changed through the administration of the food composition or health functional food of the present invention.

In the present invention, the term "food" refers to meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, There are vitamin complexes, health function (sex) 　 food, and health 　 food 　, including food additives, and all foods 　 in a normal sense.

The "healthy functional (sex) 　 food (functional food)" means a food with high medical and medical effects processed to efficiently exhibit bioregulatory functions in addition to nutrition supply. Here, "function (sex)" means to obtain a useful effect for health purposes such as regulating nutrients or physiological action with respect to the structure and function of the human body. The food composition of the present invention can be prepared by a method commonly used in the art, and at the time of manufacture, it can be prepared by adding raw materials and components commonly added in the art. In addition, if the formulation of the above 　 food is also a formulation that is recognized as a food, it can be manufactured without limitation.

The "health food" means a food having an active health maintenance or promotion effect compared to general food, and health supplement food means a food for the purpose of health supplementation. In this specification, the terms health functional food, health food, and health supplement may be used interchangeably. Specifically, the health functional food is a food prepared by adding the extract or fraction of the present invention to food materials such as beverages, teas, spices, gum, and confectionery, or manufactured by encapsulation, powdering, suspension, etc. This means that it has a specific effect, but unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long time because it uses food as a raw material. Since the 　food composition of the present invention can be consumed on a daily basis, a high effect can be expected for prevention or improvement of corona virus infection, and it can be usefully used.

In addition, the food 　 composition of the present invention may further include a physiologically acceptable carrier, the type of carrier is not particularly limited and any carrier commonly used in the art may be used. In addition, the 　food 　 composition may include additional ingredients that are commonly used in the 　food 　 composition to improve odor, taste, vision, and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, and the like may be included. Also, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr). In addition, it may include amino acids such as lysine, tryptophan, cysteine, and valine. In addition, the above 　food composition contains preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butyl hydro Loxytoluene (BHT), etc.), coloring agents (tar pigments, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasonings (MSG sodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, etc.) , sodium, etc.), flavorings (vanillin, lactones, etc.), expanding agents (alum, D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifying agents, thickening agents (flavors), filming agents, gum base agents, foam inhibitors, solvents, improving agents, etc. It may contain food additives. The additive may be selected according to the type of food and used in an appropriate amount. The extract or fraction of the present invention may be added as it is or used together with other 　food 　 or 　food ingredients, and may be appropriately used according to a conventional method.

Another aspect of the present invention for achieving the above object provides a quasi-drug composition for the prevention or improvement of coronavirus infection comprising an anti-coronavirus composition.

Specifically, the corona virus infection may refer to an infectious disease caused by a coronavirus, and specifically may refer to a coronavirus disease 19 (corona virus disease 19), but is not limited thereto.

The term "quasi-drug" as used in the present invention refers to articles with a milder action than pharmaceuticals among articles used for the purpose of diagnosing, treating, improving, alleviating, treating, or preventing diseases of humans or animals. For example, according to the Pharmaceutical Affairs Act of Korea Quasi-drugs exclude products used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases in humans and animals, and products with minor or no direct action on the human body.

The quasi-drug composition of the present invention may be prepared in the form of an ointment, lotion, spray, patch, cream, powder, suspension, gel, or filter filler, and specifically, disinfectant, wet tissue, solid soap, liquid soap, hand It may be a detergent or hand sanitizer, but is not limited thereto.

When the composition for anti-coronavirus according to the present invention is used as a quasi-drug 　 additive, the composition may be added as it is or used together with other quasi-drugs or quasi-drugs ingredients, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be suitably determined according to the purpose of use.

Another aspect of the present invention for achieving the above object provides a cosmetic composition for the prevention or improvement of coronavirus infection comprising a composition for anti-coronavirus.

Specifically, the corona virus infection may refer to an infectious disease caused by a coronavirus, and specifically may refer to a coronavirus disease 19 (corona virus disease 19), but is not limited thereto.

The above 　 cosmetics 　 compositions are solution, external ointment, cream, foam, nutrient lotion, softening lotion, pack, soft water, emulsion, makeup base, essence, lip balm, soap, liquid detergent, bath agent, hair shampoo, sunscreen cream, sun oil, Suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays may be prepared in a formulation selected from the group consisting of, but It is not limited.

In addition, the 　cosmetics 　 composition of the present application may further include one or more cosmetically acceptable carriers to be formulated in general skin 　 cosmetics, and conventional ingredients include, for example, oil, water, surfactant, humectant, lower alcohol, A thickener, a chelating agent, a colorant, a preservative, a fragrance, etc. may be appropriately mixed, but the present invention is not limited thereto. The cosmetically acceptable carrier included in the 　 cosmetic 　 composition of the present application may be variously selected according to the formulation. For example, when the formulation is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or Mixtures thereof may be used, but are not limited thereto.

When the composition for anti-coronavirus according to the present invention is used as a cosmetic composition, the composition may be added as it is or may be used in addition to the composition along with ingredients commonly used in cosmetic compositions, and may be appropriately used according to a conventional method . The mixing amount of the active ingredient may be suitably determined according to the purpose of use.

Another aspect of the present invention for achieving the above object provides a feed additive for prevention or improvement of coronavirus infection comprising a composition for anti-coronavirus.

Specifically, the corona virus infection may refer to an infectious disease caused by a coronavirus, and specifically may refer to a coronavirus disease 19 (corona virus disease 19), but is not limited thereto.

As used herein, the term "feed additive" refers to a material added in a trace amount to a feed for nutritional or specific purposes, and in the present invention, it means a material added for the purpose of preventing or improving coronavirus infection. Here, an animal is a concept including livestock and pets.

The feed additive of the present invention may further include binders, emulsifiers, preservatives, etc. added to prevent quality deterioration, and mineral preparations, mineral preparations, protective fatty acids, amino acids, vitamins, Enzymes, probiotics (lactic acid bacteria), flavoring agents, non-protein nitrogen compounds, silicates, buffers, coloring agents, extractants, oligosaccharides, etc. may be additionally included, and in addition, may further include feed mixtures, etc. It is not limited.

In the present invention Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata ), Prunus pendura ( Prunus pendula ), Prunus cerasus ( Prunus cerasus ), Prunus leveilleana ( Prunus leveilleana ) , Prunus sargentii ( Prunus sargentii ) or a mixture extract thereof is characterized in that it inhibits the entry of coronavirus into the host cell by inhibiting TMPRSS2 and ACE2 gene expression. ) can be utilized as a pharmaceutical composition, food composition, health functional food, quasi-drug, cosmetic composition or feed additive for the prevention, improvement or treatment of coronavirus infection, including.

However, effects according to an embodiment of the technology disclosed in the present specification are not limited to those mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

In order to more fully understand the drawings cited herein, a brief description of each drawing is provided.
1 is Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata ( Prunus serrulata ), Prunus pendura ( Prunus pendula ), Prunus leveilleana ( Prunus leveilleana ) and Prunus sargentii ( Prunus sargentii ) Shows the result of confirming the transcriptional inhibitory activity of TMPRSS2 mRNA of the extract (3 X 10 ^-6 g/ml).
Figure 2 Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata ( Prunus serrulata ), Prunus pendura ( Prunus pendula ), Prunus leveilleana ( Prunus leveilleana ) and Prunus sargentii ( Prunus sargentii ) Shows the result of confirming the transcriptional inhibitory activity of ACE2 mRNA of the extract (3 X 10 ^-6 g/ml).
3 shows the results of confirming the transcriptional inhibitory activity of TMPRSS2 mRNA of the Prunus cerasus extract. The results of experiments by diluting the Prunus cerasus extract extracted by the squeezing method to 1/100 (TC-2) and 1/1000 (TC-3) concentrations are shown.
Figure 4 shows the results of confirming the transcriptional inhibitory activity of ACE2 mRNA of the Prunus cerasus extract. The results of experiments by diluting the Prunus cerasus extract extracted by the squeezing method to 1/100 (TC-2) and 1/1000 (TC-3) concentrations are shown.
5 shows the results of confirming the transcriptional inhibitory activity of TMPRSS2 mRNA of a mixed extract of Prunus cerasoides and Prunus serrulata . A is a mixed extract of Prunus cerasoides and Prunus cerulata from Jeju; B is a mixed extract of Prunus cerasoides and Prunus cerulata from Gyeongsangbuk-do; C is a mixed extract of Prunus cerulata from Jeju and Prunus cerulata from Gyeongsangbuk-do; and D represents a mixed extract of Prunus cerasoides, Prunus cerulata from Jeju and Prunus cerulata from Gyeongsangbuk-do.
6 shows the results of confirming the transcriptional inhibitory activity of ACE2 mRNA of a mixed extract of Prunus cerasoides and Prunus serrulata . A is a mixed extract of Prunus cerasoides and Prunus cerulata from Jeju; B is a mixed extract of Prunus cerasoides and Prunus cerulata from Gyeongsangbuk-do; C is a mixed extract of Prunus cerulata from Jeju and Prunus cerulata from Gyeongsangbuk-do; and D represents a mixed extract of Prunus cerasoides, Prunus cerulata from Jeju and Prunus cerulata from Gyeongsangbuk-do.

Hereinafter, the present invention will be described in more detail through the following examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples only.

Example 1: Preparation of extracts of plants of the genus Prunus

Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata , Prunus pendula ( Prunus pendula ), Prunus leveilleana ( Prunus leveilleana ) and Prunus sargentii extracts from Korea It was purchased from the Plant Extract Bank and used. The extract was extracted in SD-ULTRASONIC CLEANER (SDN-900H) using 99.9% methanol as a solvent.

Prunus cerasus ( Prunus cerasus ) extract was purchased from Nature's Mammal and used. The extract is extracted by the method of squeezing.

Experimental Example 1: Confirmation of TMPRSS2 and ACE2 gene transcriptional inhibition activity of a plant alone extract of the genus Prunus

MCF-7 (human breast cancer cell line), which is known to have high TRPRSS2 and ACE2 expression, was purchased from the American Tissue Culture Collection (USA) and used to confirm the effect of inhibiting the transcriptional activity of the TMPRSS2 and ACE2 genes of the plant extract of the genus Purunus. MCF-7 was cultured in a medium composed of phenol red DMEM, 10% FBS, 1% antibiotics and 1% NEAA, and seeded in a 6-well plate at a concentration of 2.5 x 10 ⁵ cells/well.

After 24 hours, when cell confluency reached 70%, Prunus plant extract was treated in a medium composed of phenol red-free DMEM, 10% CD-FBS, 1% antibiotics and 1% NEAA. Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata , Prunus pendula ( Prunus pendula ), Prunus leveilleana ( Prunus leveilleana ) and Prunus sargentii extracts are 3 X 10 ^-6 was treated with 2.5ml at a concentration of g/ml, Prunus cerasus was diluted to 1/100 (TC-2) and 1/1000 (TC-3) concentrations of the extract extracted by the squeezing method and treated with 2.5 ml of the medium.

mRNA was extracted from the cells treated with the extract for 24 hours using Trizol. cDNA was obtained by reverse transcription reaction of a certain amount of mRNA (0.5 μg) using ReverTra Ace qPCR RT Master Mix with gDNA Remover (Toyobo). cDNA, a pair of primers capable of recognizing the cDNA of the target gene, and real-time PCR using the PCR SYBR green kit (Qiagen) reagent were performed to quantitatively measure the expression level of the corresponding gene.

In order to compensate for the technical error in the reaction in various stages, the expression level of GAPDH, a housekeeping gene, was simultaneously measured, and the ratio of the expression level of this gene to the expression level of the target gene was used for the final quantitative analysis. As primers for recognizing GAPDH, forward 5'-GGCTGAGAACGGGAAGCTTGTCAT-3' (SEQ ID NO: 1) and reverse 5'-CAGCCTTCTCCATGGTGGTGAAGA-3' (SEQ ID NO: 2) were used.

The expression levels of TMPRSS2 and ACE2 as target genes of plant extracts of the genus Purunus were measured. Forward 5'-gtgacagtggagggcctctgg-3' (SEQ ID NO: 3) and reverse 5'-cgtcaaggacgaagaccatgt-3' (SEQ ID NO: 4) were used as primers for recognizing human TMPRSS2, and forward 5'-CGAGTGGCTAATTTGAAACCAAGAA as primers for recognizing human ACE2 -3' (SEQ ID NO: 5) and reverse 5'-ATTGATACGGCTCCGGGACA-3' (SEQ ID NO: 6) were used.

Real-time PCR is approximately 40 cycles (95 °C; 15 s, 56 °C; 45 s, 72 °C; 30 s) for TMPRSS2 and approximately 50 cycles (95 °C; 15 s, 60 °C) for ACE2 C; 30 sec). After expressing each gene expression curve as a logarithm, the threshold cycle (C _T ) value was obtained mathematically, and the value obtained by substituting this value for 2-DDC _T represents the relative expression level of a specific gene. The final value of the expression level of the gene was calculated by dividing the expression level of a specific gene by the expression level of the housekeeping gene.

As a result, as shown in FIGS. 1 to 4, it was confirmed that the expression of TMPRSS2 and ACE2 was significantly inhibited in the experimental group treated with the plant extract of the genus Purunus compared to the control group.

Experimental Example 2: Confirmation of TMPRSS2 and ACE2 gene transcriptional inhibition activity of Prunus genus plant mixed extract

In the same manner as in Experimental Example 1, a mixed extract of Prunus cerasoides and Jeju Prunus cerulata; A mixed extract of Prunus cerasoides and Prunus cerulata from Gyeongsangbuk-do; A mixed extract of Prunus cerulata from Jeju and Prunus cerulata from Gyeongsangbuk-do; and TMPRSS2 and ACE gene transcription inhibitory activity of the mixed extracts of Prunus cerasoides, Prunus cerulata from Jeju and Prunus cerulata from Gyeongsangbuk-do.

As a result, as shown in FIGS. 5 and 6 , it was confirmed that the expression of TMPRSS2 and ACE2 was significantly suppressed in the experimental group treated with the mixed extract compared to the control group.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the following claims and their equivalents.

<110> Sookmyung Women's university industry-academic cooperation foundation <120> Composition for anti-coronavirus containing extract of Prunus genus <130> KPA01527 <160> 6 <170> KoPatentIn 3.0 <210> 1 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> artificial sequence <400> 1 ggctgagaac gggaagcttg tcat 24 <210> 2 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> artificial sequence <400> 2 cagccttctc catggtggtg aaga 24 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> artificial sequence <400> 3 gtgacagtgg agggcctctg g 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> artificial sequence <400> 4 cgtcaaggac gaagaccatg t 21 <210> 5 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> artificial sequence <400> 5 cgagtggcta atttgaaacc aagaa 25 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> artificial sequence <400> 6 attgatacgg ctccgggaca 20

Claims (11)

Prunus ( Prunus ) Comprising an extract of a plant of the genus as an active ingredient, anti-coronavirus composition. The method of claim 1,
The Prunus genus plants are Prunus cerasoides ( Prunus cerasoides ), Prunus serrulata ( Prunus serrulata ), Prunus pendura ( Prunus pendula ), Prunus cerasus ( Prunus cerasus ), Prunus rebaileana ( Prunus leveilleana ), Prunus sargentii ( Prunus sargentii ), or a combination thereof, the composition. The composition of claim 1, wherein the composition inhibits the expression of TMPRSS2 (transmembrane protease serine subtype 2) and ACE2 (angiotensin-converting enzyme 2) genes. The composition of claim 1 , wherein the anti-virus inhibits entry of the corona virus into a host cell. A pharmaceutical composition for the prevention or treatment of coronavirus infection, comprising the composition of any one of claims 1 to 4. A food composition for the prevention or improvement of coronavirus infection, comprising the composition of any one of claims 1 to 4. Health functional food for the prevention or improvement of coronavirus infection comprising the food composition of claim 6. A quasi-drug composition for the prevention or improvement of coronavirus infection, comprising the composition of any one of claims 1 to 4. 9. The method of claim 8,
The quasi-drug composition is a quasi-drug composition used in the manufacture of disinfectants, wipes, solid soap, liquid soap, hand sanitizer or hand sanitizer. A cosmetic composition for the prevention or improvement of coronavirus infection, comprising the composition of any one of claims 1 to 4. A feed additive for the prevention or improvement of coronavirus infection, comprising the composition of any one of claims 1 to 4.

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