KR20200124959A - ANTIVIRAL COMPOSITION COMPRISING COMPOUND ISOLATED FROM Castanea crenata - Google Patents

Abstract

The present invention relates to an antiviral composition containing a chestnut bur-derived compound as an active component. The composition can effectively fight against Picornaviridae (for example, HRV1B, CVB3, etc.) or Orthomyxoviridae (for example, PR8, etc.) which is a virus that causes respiratory diseases, thereby being able to be usefully used as a pharmaceutical composition, health functional food, skin external preparation, and quasi-drug.

Description

Antiviral composition containing a compound derived from chestnut as an active ingredient{ANTIVIRAL COMPOSITION COMPOUND ISOLATED FROM Castanea crenata}

The present invention relates to an antiviral composition comprising a compound derived from chestnut pine as an active ingredient.

Viral infection is one of the most common diseases, and more than 60% of acute respiratory diseases are caused by viruses. Among about 200 kinds of viral respiratory diseases, the most common and important infectious viruses in humans are influenza virus, rhinovirus and coxsackie virus. Human rhinovirus 1B (HRV1B) belongs to the genus Enterovirus of the family Picornavirus. HRV is the leading cause of colds, and colds caused by this virus can lead to pneumonia and bronchiolitis. Coxsackievirus B3 (CVB3) of the Picornavirus family is a virus that can cause myocarditis and can cause dilated cardiomyopathy. Myocarditis caused by this virus can lead to death in infants and young children, and may lead to end-stage congestive heart failure in adults. In addition, influenza virus is an RNA virus of the Orthomyxovirus family, which causes infectious respiratory diseases, and causes a worldwide flu outbreak every year, causing infections at a high rate, especially among the elderly, infants, and patients with various respiratory or internal medical diseases. Raises. Therefore, much effort has been made for the prevention and treatment of diseases caused by such various viral infections, but a method that can be called a clear solution is still insufficient.

The present invention is to provide an antiviral pharmaceutical composition containing as an active ingredient one or more compounds selected from the following i) and ii) or a pharmaceutically acceptable salt thereof:

I) Camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OZp -coumaroyl]-β-D-glucopyranoside), astragarin, tiliroside, cis-tiliroside, camphorol-3-O-[2"-OEp-cumaro Yl]-β-D-glucopyranoside (kaempferol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3-O-[3",6"-di- O-Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3- O-[2",6"-di-O-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OEp-coumaroyl]-β -D-glucopyranoside), camperol-3-O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[ 4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) and camphorol-3-O-[3"-acetyl-2",6"-di-Ep-coumaro One or more selected from the group consisting of]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) Flavonoid-based compounds comprising a; And

Ii) A cycloartane type compound containing castaartancrenoic acid D or castaartancrenoic acid E.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems that are not mentioned will be clearly understood by those skilled in the art from the following description.

The present invention provides an antiviral pharmaceutical composition containing as an active ingredient one or more compounds selected from the following i) and ii) or a pharmaceutically acceptable salt thereof:

I) Camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OZp -coumaroyl]-β-D-glucopyranoside), astragarin, tiliroside, cis-tiliroside, camphorol-3-O-[2"-OEp-cumaro Yl]-β-D-glucopyranoside (kaempferol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3-O-[3",6"-di- O-Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3- O-[2",6"-di-O-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OEp-coumaroyl]-β -D-glucopyranoside), camperol-3-O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[ 4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) and camphorol-3-O-[3"-acetyl-2",6"-di-Ep-coumaro One or more selected from the group consisting of]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) Flavonoid-based compounds comprising a; And

Ii) A cycloartan-based compound containing castaartancrenoic acid D or castaartancrenoic acid E.

The compound can be isolated from chestnut extract.

The composition may be for combating a virus that causes respiratory diseases.

The virus may be of the family Picornavirus (Picornaviridae) or family of Orthomyxoviridae (Orthomyxoviridae).

The Picornavirus family (Picornaviridae) may include human rhinovirus 1B (HRV1B) or coxsackie virus B3 (CVB3), and the Orthomyxoviridae family may include influenza virus (PR8).

In one embodiment of the present invention, there is provided an antiviral health functional food containing one or more compounds selected from i) and ii) or a pharmaceutically acceptable salt thereof as an active ingredient.

In another embodiment of the present invention, there is provided an antiviral external preparation for skin containing one or more compounds selected from i) and ii) or a pharmaceutically acceptable salt thereof as an active ingredient.

In another embodiment of the present invention, there is provided an antiviral quasi-drug containing one or more compounds selected from i) and ii) or a pharmaceutically acceptable salt thereof as an active ingredient.

In another embodiment of the present invention, a novel compound represented by the following formula 1 is provided:

[Formula 1]

.

.

The compound can be isolated from chestnut extract.

The present invention relates to an antiviral composition comprising a compound derived from chestnut as an active ingredient, wherein the composition is a virus that causes respiratory diseases, such as Picornaviridae (e.g., HRV1B, CVB3, etc.) or Orthomyxovirus ( Orthomyxoviridae) (for example, PR8, etc.) can be effectively opposed, and can be usefully used as pharmaceutical compositions, health functional foods, skin external preparations and quasi-drugs.

1 shows the chemical structures of compounds (1)-(5), (8), (9) and (12)-(16).
2(a) shows NMR spectral data for compounds (1)-(3), FIG. 2(b) shows NMR spectral data for compounds (4)-(5), and FIG. 2(c) ) Shows the NMR spectral data for compound (16).
Figure 3 shows the major HMBC and COSY correlation of compounds (1), (3), (4) and (16).
Figure 4 shows the major NOESY correlations of compounds (1), (2), (4) and (5).

In a continuing attempt to discover natural substances exhibiting antiviral effects, a phytochemical study of chestnut chestnuts of Castanea crenata Siebold & Zucc. Chestnut ( C. crenata ), a type of chestnut tree native to Korea and Japan, has been used in traditional medicine to treat gastroenteritis, bronchitis and reflux. Chestnut treatment produces two wastes: chestnut and bark. While the bark is separated during the peeling process and used as fuel, chestnuts generally remain in the forest, promoting the reproduction of larvae and damaging crops. As an herbal material, chestnut chestnuts called involucre have been used to treat boils, whooping cough and bronchitis. Chestnut ( C. crenata ) Previous phytochemical studies on chestnuts have identified various chemical components such as tannins, phenolic acids, flavonoids, coumarins, phenylpropanoids and steroids. In addition, it has been reported that chestnut ( C. crenata ) extract showed inhibitory activity against human immunodeficiency virus (HIV)-1 protease. Therefore, the phytochemical study of chestnut chestnuts is significant as the discovery of a previously unknown natural antiviral source.

A total of 16 types of cycloartane triterpene compounds and flavonoid glycosides were isolated from chestnut ( C. crenata ) chestnut. The structure was described based on spectroscopic analysis of 1D and 2D NMR and MS data. The isolated compounds were evaluated for their biological activity against HRV1B-, CVB3- and PR8-infected Vero or HeLa cells. Most camphorol derivatives showed statistically significant antiviral activity against HRV1B-infected cells. Of the compounds tested, camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside showed the most consistent and effective antiviral activity against all infections. Indicated.

Hereinafter, the present invention will be described in detail.

The present invention provides an antiviral composition containing as an active ingredient at least one compound selected from the following i) and ii) or a pharmaceutically acceptable salt thereof:

I) Camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OZp -coumaroyl]-β-D-glucopyranoside), astragarin, tiliroside, cis-tiliroside, camphorol-3-O-[2"-OEp-cumaro Yl]-β-D-glucopyranoside (kaempferol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3-O-[3",6"-di- O-Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3- O-[2",6"-di-O-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OEp-coumaroyl]-β -D-glucopyranoside), camperol-3-O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[ 4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) and camphorol-3-O-[3"-acetyl-2",6"-di-Ep-coumaro One or more selected from the group consisting of]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) Flavonoid-based compounds comprising a; And

Ii) A cycloartan-based compound containing castaartancrenoic acid D or castaartancrenoic acid E.

That is, the antiviral composition according to the present invention may contain one or more compounds selected from i) flavonoid-based compounds and ii) cycloartane-based compounds or a pharmaceutically acceptable salt thereof as an active ingredient.

The compound may be isolated from the chestnut extract. Specifically, the compound may be isolated from the fraction of the chestnut extract. First, the chestnut extract may be extracted with water, a lower alcohol of C1 to C4, or a mixture thereof, and the chestnut extract is preferably extracted with methanol. However, it is not limited thereto. In addition, the fraction of the chestnut extract may be obtained by suspending the chestnut pine tree extract in distilled water and then fractionating the chestnut pine tree extract from a non-polar solvent using dichloromethane and ethyl acetate.

First, i) the flavonoid compound is camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2", 6"-di-OZp-coumaroyl]-β-D-glucopyranoside) (compound (14)), astragarin (compound (8)), tiliroside (compound (9)), cis- Cis-tiliroside (compound (10)), camperol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2 "-OEp-coumaroyl]-β-D-glucopyranoside) (compound (11)), camphorol-3-O-[3",6"-di-O-Ep-coumaroyl]-β-D-glu Copyranoside (kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside) (compound (12)), camphorol-3-O-[2",6" -D-O-Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OEp-coumaroyl]-β-D-glucopyranoside) (compound ( 13)), camphorol-3-O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[4" -acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) (compound (15)) and camphorol-3-O-[3"-acetyl-2",6"-di- Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) (Compound ( 16)), and may include one or more selected from the group consisting of, among which camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside) (compound (14)) It is preferable in that it can effectively fight a wide range of viruses, but is not limited thereto.

In addition, ii) the cycloartan-based compound may include castaartancrenoic acid D (compound (4)) or castaartancrenoic acid E (castaartancrenoic acid E) (compound (5)). have.

The composition may be for combating a virus that causes respiratory disease, and thus, it is possible to ultimately prevent, improve or treat respiratory diseases caused by viruses and accompanying infections. At this time, the respiratory disease is preferably emphysema, chronic bronchitis, bronchial adenoma, isolated pulmonary nodule, pulmonary tuberculosis, empyema, lung abscess, cold, flu, and histocytosis of the lung, but is not limited thereto.

The virus may be Picornavirus family (Picornaviridae) or Orthomyxoviridae family (Orthomyxoviridae), specifically, the Picornavirus family (Picornaviridae) includes human rhinovirus 1B (HRV1B) or Coxsackie virus B3 (CVB3) And, the Orthomyxoviridae may include influenza virus (PR8).

When the composition is a pharmaceutical composition, each of the pharmaceutical compositions is in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injectable solutions according to a conventional method. It may be formulated and used, and may include an appropriate carrier, excipient, or diluent commonly used in the preparation of pharmaceutical compositions for formulation.

As the carrier or excipient or diluent, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, undecided And various compounds or mixtures including vaginal cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

In the case of formulation, it can be prepared using diluents or excipients such as fillers, weight agents, binders, wetting agents, disintegrants, and surfactants that are usually used.

Solid preparations for oral administration may be prepared by mixing the compound with at least one excipient such as starch, calcium bonate, sucrose or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used.

Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. may be included. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous agents, suspensions, emulsions, lyophilized formulations, and suppositories. As the non-aqueous solvent and suspension, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerol gelatin, and the like can be used.

The preferred dosage of the pharmaceutical composition varies depending on the patient's condition, weight, degree of disease, drug form, administration route and duration, but may be appropriately selected by those skilled in the art. However, for a desirable effect, it may be administered at 0.0001 to 2,000 mg/kg per day, preferably 0.001 to 2,000 mg/kg. Administration may be administered once a day, or may be divided several times. However, the scope of the present invention is not limited by the dosage.

The pharmaceutical composition may be administered to mammals such as mice, mice, livestock, and humans by various routes. All modes of administration can be administered by, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

On the other hand, when the composition is a health functional food, in the health functional food, when the compound is used as an additive of a health functional food, it can be added as it is or used with other foods or food ingredients, according to a conventional method. Can be used appropriately. The mixing amount of the active ingredient can be appropriately determined according to each purpose of use, such as prevention, health or treatment.

The formulation of the health functional food may be in the form of powders, granules, pills, tablets, capsules, as well as general foods or beverages.

The type of food is not particularly limited, and examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , Various soups, beverages, teas, drinks, alcoholic beverages and vitamin complexes, and may include all foods in the usual sense.

In general, when preparing food or beverage, the compound may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on 100 parts by weight of raw materials. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of health control, the amount may be less than or equal to the above range, and the present invention uses natural substances, so there is no problem in terms of safety. Can also be used as.

Among the health functional foods, beverages may contain various flavoring agents or natural carbohydrates as an additional component, like ordinary beverages. The natural carbohydrates described above may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the beverage.

In addition to the above, the health functional food includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, It may contain carbonating agents used in carbonated beverages. In addition, the health functional food may contain pulp for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These ingredients may be used independently or in combination. The ratio of these additives is not limited, but it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food of the present invention.

On the other hand, when the composition is an external preparation for skin, suitable formulations used for the external preparation for the skin include, for example, solutions, gels, solid or paste dry products, emulsions obtained by dispersing an oil phase in an aqueous phase, suspension, microemulsion, microcapsules. , Microgranular or ionic (liposome), nonionic vesicle dispersant, cream, skin, lotion, powder, ointment or spray may be provided. In addition, it may be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

In addition to the compound, the external preparation for skin is a fatty substance, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, ionic or non Ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in external preparations for skin It may contain adjuvants commonly used in the field of dermatology such as. In addition, the ingredients may be introduced in an amount generally used in the field of dermatology.

On the other hand, when the composition is a quasi-drug, the compound may be added as it is or may be used with other quasi-drug or quasi-drug components, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use.

The quasi-drug is not limited thereto, but may be preferably a disinfectant cleaner, a shower foam, a gagrin, a wet tissue, a detergent soap, a hand wash, a humidifier filler, a mask, an ointment, a patch, or a filter filler.

In addition, it provides a novel compound represented by the following formula 1:

[Formula 1]

.

.

The compound is a specific new compound having antiviral activity among compounds derived from chestnut pine. Specifically, the novel compound represented by Formula 1 is "Compound (16): kaempferol-3-O-[3" in Examples to be described later. -acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside".

The compound may be isolated from the chestnut extract. Specifically, the compound may be isolated from the fraction of the chestnut extract. First, the chestnut extract may be extracted with water, a lower alcohol of C1 to C4, or a mixture thereof, and the chestnut extract is preferably extracted with methanol. However, it is not limited thereto. In addition, the fraction of the chestnut extract may be obtained by suspending the chestnut pine tree extract in distilled water and then fractionating the chestnut pine tree extract from a non-polar solvent using dichloromethane and ethyl acetate.

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention. However, the following examples are provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Normal

Chemical shifts are expressed in parts per million from TMS. ¹ H and ¹³ C NMR spectra were run at 800 and 200 MHz measured with an Agilent 800-MR-NMR spectrometer, respectively. Data processing was performed with the MestReNova ver.6.0.2 program. HRESI-MS experiments were performed using an Agilent 6550 iFunnel QTOF LC/MS system. Optical rotation was measured on a Jasco DIP-370 automatic polarimeter. Preparative HPLC was performed on an Agilent 1260 HPLC system. Column chromatography (CC) was performed with silica gel (Kieselgel 60, 70-230 mesh and 230-400 mesh, Merck) and RP-silica gel (150 μm, Fuji Silysia Chemical). Thin layer chromatography (TLC) was performed on pre-coated silica gel 60 F254 (0.25 mm, Merck) and RP-18 F254S plates (0.25 mm, Merck) plates. The FT-IR spectrum (KBr) was recorded on a Nicolet 380 spectrometer.

Plant material

During the dry season of April 2014, chestnuts of Castanea crenata Siebold & Zucc. (Oakaceae) were collected from a Seokjang farm in Deoksan-myeon, Jincheon-gun, Chungcheongbuk-do, Korea. I did.

Extraction and separation

Crushed chestnut ( C. crenata ) chestnut (9.7 kg) was sonicated for 15 hours and extracted with 80% methanol (20 L×3 times) to obtain 532 g extract after evaporation of the solvent. The extract was suspended in water and partitioned successively with dichloromethane and ethyl acetate to obtain dichloromethane (CB1, 123.21 g), ethyl acetate (CB2, 75.37 g) and water (0.15 g) fractions after removal of the solvent under vacuum. . The CH ₂ Cl ₂ residue was applied to silica gel CC with a gradient of CHCl ₃ :MeOH (40:1→1:1,v/v) to obtain 6 sub-fractions (CB1A-CB1F). The CB1C fraction was applied to the silica gel CC eluted with a CHCl ₃ :MeOH (10:1,v/v) solvent system to obtain a fraction smaller than CB1C1-CB1C6. CB1C3 was applied to HPLC using a J'sphere ODS H-80 250 mm × 20 mm column eluted with 43% MeCN at a flow rate of 3 mL/min to obtain Compound (15) (9.6 mg) and Compound (16) ( 12 mg), while CB1C4 yielded compounds (1)-(3) (12.6, 4.3, and 5.9 mg) by HPLC eluting with 40% aqueous MeCN. Additionally, the CB1C5 fraction was also subjected to HPLC eluting with 65% aqueous MeCN to give compound (4) (3.3 mg) and compound (5) (4.5 mg). The EtOAc soluble fractions were divided into 6 sub-fractions (CB2A-CB2F) using CHCl ₃ :MeOH gradient eluate (40:1 → 1:1, v/v) on silica gel column chromatography. Using silica gel CC eluted with CHCl ₃ :MeOH (5:1,v/v), the CB2C fraction was mixed with less than 4 fractions (CB2C1-CB2C4), compound (6) (41.3 mg), compound (17 ) (24.2 mg) was allowed to be obtained. The CB2C1 fraction was applied to RP-silica gel CC eluted with MeOH: H ₂ O (1:1,v/v) to obtain compound (7) (6.6 mg), compound (8) (81.3 mg), compound (9) ( 5.8 mg) and compound (10) (11.6 mg) were provided. The CB2C2 fraction was further purified by HPLC using 25% aqueous MeCN to obtain compound (11) (7.0 mg), compound (12) (8.9 mg), compound (13) (3.2 mg) and compound (14) (4.8 mg ) Was obtained.

-Castaartancrenoic acid A (Compound (1))

Yellow amorphous powder; [ α ] _D ²⁰ : +10.0 (MeOH, c=2.0); IR (KBr) ν _max : 3300, 1653, 1445, 1165, 1017 cm ^-1 ; C ₂₇ H ₄₂ O ₆ , HR-ESI-MS m/z: 461.2903 [MH] ^- (calculated for C ₂₇ H ₄₁ O ₆ , 461.2909); ¹ H (CD ₃ OD, 800 MHz) and ¹³ C NMR (CD ₃ OD, 200 MHz) Spectroscopy data subject, see Table 1

-Castaartancrenoic acid B (Compound (2))

Yellow amorphous powder; [ α ] _D ²⁰ : +18.0 (MeOH, c=2.0); IR (KBr) ν _max : 3340, 1708, 1448, 1114, 1021 cm ^-1 ; C ₂₇ H ₄₄ O ₅ , HR-ESI-MS m/z: 447.3108 [MH] ^- (calculated for C ₂₇ H ₄₃ O ₅ , 447.3116); ¹ H (CD ₃ OD, 800 MHz) and ¹³ C NMR (CD ₃ OD, 200 MHz) Spectroscopy data subject, see Table 1

-Castaartancrenoic acid C (Compound (3))

Yellow amorphous powder; [ α ] _D ²⁰ : +55.0 (MeOH, c=2.0); IR (KBr) ν _max : 3345, 1650, 1420, 1114, 1021 cm ^-1 ; C ₂₆ H ₄₁ O ₅ ,HR-ESI-MS m/z: 433.2950 [MH] ^- (calculated for C ₂₆ H ₄₁ O ₅ , 433.2959; ¹ H (CD ₃ OD, 800 MHz) and ¹³ C NMR (CD ₃ OD, 200 MHz) for spectral data, see Table 1

-Castaartancrenoic acid D (compound (4))

Yellow amorphous powder; [ α ] _D ²⁰ : +24.2 (MeOH, c=2.0); IR (KBr) ν _max : 3363, 1707, 1437, 1010, 950 cm ^-1 ; C ₃₀ H ₄₈ O ₅ , HR-ESI-MS m/z: 487.3422 [MH] ^- (calculated for C ₃₀ H ₄₇ O ₅ , 487.3429); ¹ H (pyridine, 800 MHz) and ¹³ C NMR (pyridine, 200 MHz) spectroscopy data subject, see Table 2

-Castaartancrenoic acid E (Compound (5))

Yellow amorphous powder; [ α ] _D ²⁰ : +40.0 (MeOH, c=2.0); IR (KBr) ν _max : 3440, 1672, 1437, 1093, 1012 cm ^-1 ; C ₃₀ H ₄₈ O ₅ , HR-ESI-MS m/z: 487.3429 [MH] ^- (calculated for C ₃₀ H ₄₇ O ₅ , 487.3429); ¹ H (pyridine, 800 MHz) and ¹³ C NMR (pyridine, 200 MHz) spectroscopy data subject, see Table 2

-Camperol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl- 2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) (compound (16))

Yellow amorphous powder; [ α ] _D ²⁰ : +25.0 (MeOH, c=2.0)); IR (KBr) ν _max : 3320, 1653, 1605, 1411 cm ^-1 ; C ₄₁ H ₃₄ O ₁₆ , HR-ESI-MS m/z: 781.1789 [MH] ^- (calculated for C ₄₁ H ₃₃ O ₁₆ , 781.1769); ¹ H (CD ₃ OD, 800 MHz) and ¹³ C NMR (CD ₃ OD, 200 MHz) Spectroscopic data subject, see Table 3

Cell culture

HRV1B, CVB3 and PR8 were obtained from the Korea Centers for Disease Control and Prevention, and were proliferated at 37°C in HeLa and Vero cells (ATCC, Manassas, VA, USA). HRV1B and PR8 infected HeLa cells, and CVB3 infected Vero cells. These cells were maintained in minimal essential medium (MEM) supplemented with 10% fetal bovine serum (FBS) and 0.01% antibiotic-antifungal solution. Antibiotic-antifungal solution, trypsin-EDTA, FBS and MEM were purchased from Gibco BRL (Invitrogen Life Technologies, Karlsruhe, Germany). Tissue culture plates were purchased from Falcon (BD Biosciences, San Jose, CA, USA). SRB was purchased from Sigma-Aldrich (St. Louis, MO, USA). Cells were cultured in MEM supplemented with a solution of 10% FBS and 0.01% antibiotic-antifungal agent at 37° C. in a humidified atmosphere of 5% CO ₂ . Cells were detached using trypsin-EDTA. Before the cells were cultured with the isolated compound, subculture was performed 5 times.

Antiviral assay

Antiviral activity was analyzed by employing the SRB method using cytopathic effect (CPE). Briefly, Vero and HeLa cells were applied using MEM supplemented with 10% FBS and 0.01% antibiotic-antifungal solution on a 96-well culture plate and incubated for 24 hours. The next day, the medium was removed, and the cells were washed with 100 μL of PBS (Invitrogen Life Technologies). Then, a diluted virus suspension (0.09 mL) containing a 50% cell culture infection dose of the virus (CCID ₅₀ ) was added to Vero cells to produce 50% CPE within 48 hours post infection. Next, MEM (0.1 mL) containing 10 μM of the compound was added. Culture plates were incubated at 5% CO ₂ and 37° C. for 2 days until 50% CPE was achieved. Then, the 96-well plate was washed once with 200 mL of PBS. Cold 70% acetone (100 μL) was added to each well and left at room temperature for 30 minutes. Then the SRB solution was removed and the plate was washed 5 times with 1% acetic acid in water before oven-drying at 55°C. Then, SRB was dissolved with 10 mM unbuffered Tris-base (Sigma) solution (100 μL). After 30 minutes, absorbance was measured at 540 nm using a VERSAmax microplate reader (Molecular Devices, Palo Alto, CA, USA) having a reference absorbance at 620 nm. HRV1B-, CVB3- and PR8- each test compound in infected cells The antiviral activity of was calculated as a percentage of the cell viability of the corresponding untreated control.

Results and discussion

Chestnut ( C. crenata ) The methanol extract of chestnut was suspended in water, and successively distributed with dichloromethane and ethyl acetate to obtain three layers. Using various chromatography and separation techniques, 6 new compounds and 11 known compounds were isolated (Figure 1). This structure was explained by broad spectrum spectroscopy methods including 1D and 2D NMR experiments and by HR-ESI-MS analysis. The remaining 11 known compounds were compared to those of previously reported literature data, and stachyanthuside B (compound (6)), 3,3'-di-methylelazic acid 4'-O-β-D -Xylopyranoside (3,3'-di-methylellagic acid 4'-O-β-D-xylopyranoside) (Compound (7)), astragalin (Compound (8)), Tyriroside ( tiliroside) (compound (9)), camphorol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside (Kaempferol-3-O-[2"-OEp-coumaroyl] -β-D-glucopyranoside) (compound (11)), camphorol-3-O-[2",6"-di-O-Ep-coumaroyl]-β-D-glucopyranoside (Kaempferol- 3-O-[2",6"-di-OEp-coumaroyl]-β-D-glucopyranoside) (compound (13)), camphorol-3-O-[2",6"-di-OZp-coumaroyl Royl]-β-D-glucopyranoside (Kaempferol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside) (compound (14)), camphorol-3 -O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (Kaempferol-3-O-[4"-acetyl-2",6" -di-Ep-coumaroyl]-β-D-glucopyranoside) (compound (15)), cis-tiliroside (compound (10)), camphorol-3-O-[3",6 "-D-O-Ep-Coumaroyl]-β-D-glucopyranoside (Kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside) (compound (12)) and helicrysoside (compound (17)).

Was obtained for compound (1) as a yellow amorphous powder, its molecular formula was m / z 461.2903 HR-ESI- MS [MH] from (C ₂₇ H ₄₁ O ₆ Calcd, 461.2909 ^for) by ion C ₂₇ H ₄₂ O _It was decided as ₆ . The ¹ H NMR spectrum of compound (1) shows two cyclopropane methylene protons at δ _H 0.21 (d, J = 4.4 Hz) and 0.70 (d, J = 4.4 Hz), δ _H 3.91 (dd, J = 4.2, 11.6). ) And 2 oxymethine protons at 3.43 (ddd, J = 6.4, 10.2, 12.0 Hz), 1 secondary methyl group at δ _H 0.81 (d, J = 6.2 Hz) and δ _H 0.86 (s), 0.91 (s ), and at 1.00 (s), the signals of the three tertiary methyl groups were shown. These were similar to those of other cycloartane triterpenes. The ¹³ C NMR and DEPT spectra of compound (1) showed a signal of 27 carbons including 4 methyl, 10 methylene, 6 methine and 7 quaternary carbons (Fig. 2(a)), which is a tree It was assigned as a norcycloartane triterpene. The NMR spectroscopy data of compound (1) was similar to that of norquadrangularic acid A, except for the presence of a hydroxy group at C-7 and loss of a hydroxy group at C-1. H-29 (δ _H 1.00) and C-3 (δ _C 76.2)/C-4(δ _C 55.4)/C-5(δ _C The HMBC correlation between 43.6)/C-28(δ _C 180.6) suggested the positions of the hydroxy group at C-3 and the carboxyl group at C-28. H-7(δ _H 3.43) between H-6 (δ _H 1.28) and C-7 (δ _C 70.6) and H-22 (δ _H 1.74)/H-23 (δ _H 2.12) and C-24 (δ _C The HMBC correlation between 178.4) confirmed the positions of the hydroxy group at C-7 and the carboxyl group at C-24 (Fig. 3). The large adjacent coupling constants for H _α -2 and H-3 (J _{Hα -2/H-3} =11.6 Hz) indicated their trans biaxial orientation. In addition, the axial directions of H-7 and H-8 were deduced by their large coupling constant values (J _{H -7/H-8} =7.6 Hz). Moreover, H-3 (δ _H 3.91) and H-5 (δ _H 2.03); H-29 (δ _H 1.00) and H _β -19 (δ _H 0.70); The observation of the NOESY correlation between H-7 (δ _H 3.43) and H-30 (δ _H 0.86) is β, which is the hydroxyl at C-3 and C-7. As the timing and α, the arrangement of the carboxyl groups in C-4 was proposed (Fig. 4). Based on the above data, the structure of compound (1) is 3β,7β-dihydroxy-24-norcycloartane-24,28-dioic acid (3β,7β-dihydroxy-24-norcycloartane-24,28-dioic acid) and castaartancrenoic acid A.

Compound (2) HR-ESI-MS is [MH] in the (calculated, 447.3116 for _{C 27 H 43 O 5) m} / z 447.3108 for ^- indicating ion, suggested a molecular formula of C ₂₇ H ₄₄ O _5. The ¹ H NMR spectrum of compound (2) showed a signal of a cycloartane triterpene. The NMR data showed that the structure of compound (2) was similar to that of compound (1), except that there was no carbonyl group at C-24. The COSY experiment established the position of the hydroxymethylene group in C-23 by establishing a spin system of -CH ₂ -CH ₂ -CH-CH(CH ₃ )-CH ₂ -CH ₂ -CH ₂ -. Based on the above data, the structure of compound (2) is 3β,7β,24-trihydroxy-24-norcycloartane-28-oic acid (3β,7β,24-trihydroxy-24-norcycloartane-28-oic acid) and castaartancrenoic acid B.

The molecular formula of Compound (3) HR-ESI-MS [MH] in the (calculated, 433.2959 for _{C 26 H 41 O 5) m} / z 433.2905 - was confirmed by ion as C ₂₆ H ₄₂ O _5. In addition, ¹ H NMR and ¹³ C NMR spectra of compound (3) showed typical signals of cycloartane triterpenes: methylene of the cyclopropane ring, 2 oxymethines, 1 oxymethylene, 3 tertiary methyl groups, and 1 secondary methyl group. Additionally, analysis of the NMR data indicated that the structure of compound (3) was similar to that of compound (2), except that the pentanol group at C-20 was replaced with a butanol group. H-22(δ _H 1.66) to C-23 (δ _C The HMBC correlation up to 60.1) confirmed the position of the hydroxy group at C-23 (Fig. 3). Based on the above evidence, the structure of compound (3) is 23-norcy-cloartane-28-oic acid and castaartancrenoic acid C. It was named.

Compound 4 was isolated as a yellow amorphous powder, its molecular formula was _{m / z 487.3422 (C 30 H} 48 O Calcd _{5, 487.3429) HR-ESI-} MS [MH] in ^- by ion C ₃₀ H ₄₈ O _It was identified as ₅ . ^{The 1} HNMR spectrum suggested that compound (4) is a cycloartane triterpene: methylene proton of the cyclopropane ring at δ _H 0.36 (d, J = 4.2 Hz) and 1.03 (m), δ _H 3 oxymethine protons at 3.99 (d, J = 10.2 Hz), 4.43 (m) and 4.78 (dd, J = 3.8, 11.8 Hz). Additionally, with one secondary methyl group at δ _H 1.06 (d, J=6.0 Hz), exo-olefin protons at δ _H 5.05 (s) and 5.28 (s), δ _H 1.12 (s), 1.14 (s) , At 1.71(s) and 2.00(s), four tertiary methyl groups were seen. The ¹³ C NMR and DEPT spectra of compound (4) showed the presence of 30 carbons including 5 methyls, 11 methylenes, 8 methine and 6 quaternary carbons (FIG. 2(b)). The NMR data of compound (4) was similar to that of combretic acid B, except that C-7 was substituted with a hydroxy group instead of an acetyl group. The HMBC correlation was similar to that of compounds (1)-(3) (Fig. 3). Additionally, H-23 (δ _H 2.00) and C-22 (δ _C 32.5)/C-24(δ _C 76.1)/C-25(δ _C 148.8), H-24(δ _H 4.43) and C-26 (δ _C 110.9)/C-27(δ _C The HMBC correlation between 17.5) proved the position of the hydroxy group at C-24 and the exo-olefin group at C-26 (Fig. 3). The arrangement of the hydroxy groups in C-24 as α is NOESY correlation H-21 (δ _H 1.06) and H-22 (δ _H 1.22); Hβ-23 (δ _H 2.00) and H-24 (δ _H 4.43) (Fig. 4). As a result, the structure of compound (4) is 3β,7β,24α-trihydroxycycloart-25,26-en-28-oic acid (3β,7β,24α-trihydroxycycloart-25,26-ene-28- oic acid) and castaartancrenoic acid D.

The molecular formula of the compound (5) HR-ESI-MS [MH] in the (calculated, 487.3429 for _{C 30 H 47 O 5) m} / z 487.3429 - was found to be by ion C ₃₀ H ₄₈ O _5. The ¹ H and ¹³ C NMR spectroscopy data of compound (5) were similar to that of compound (4) except for the arrangement of the hydroxy group at C-24. The β configuration of the hydroxy group in C-24 is H-21 (δ _H 1.05) and H _α -22 (δ _H 1.74); H _α -22 (δ _H 1.74) and H-24 (δ _H 4.44) was determined on the basis of the NOESY correlation. Additionally, the arrangement of C-24 can be determined by comparing the ¹³ C NMR signals of C-24 to C-27. Quadrangularic acid M (quadrangularic acid M) having a 24β hydroxy group is δ _C 75.6 (C-24), 149.6 (C-25), 110.0 (C-26) and 18.2 (C-27), while 24-Epiquadrangularic acid M (24-Epiquadrangularic acid M) having a 24α hydroxyl group ) Showed δ _C 76.1 (C-24), 149.6 (C-25), 110.4 (C-26), and 17.7 (C-27). The ¹³ C-NMR chemical shift from C-24 to C-27 in compounds (4) and (5) is δ _C 76.1 (C-24), 148.8 (C-25), 110.9 (C-26) and 17.5 (C-27); It was 75.6 (C-24), 149.0 (C-25), 110.4 (C-26) and 17.8 (C-27), and the alignment at C-24 was determined to be 24α and 24β, respectively. Based on the above data, the structure of compound (5) is 3β,7β,24β-trihydroxycycloart-25,26-en-28-ic acid (3β,7β,24β-trihydroxycycloart-25,26-ene- 28-oic acid) and castaartancrenoic acid E.

Compound 16 was isolated as a yellow amorphous powder, its molecular formula was _{m / z 781.1799 (C 41 H} 33 O Calcd _{16, 781.1769) HR-ESI-} MS [MH] in ^- by ion C ₄₁ H ₃₄ O _It was decided to be ₁₆ . The ¹ H NMR spectrum of compound (16) was δ _H 6.21 (d, J=16.0 Hz) and 7.30 (d, J=16.0 Hz); At 6.00 (d, J=16.0 Hz) and 7.58 (d, J=16.0 Hz), the signals of the two trans-coumaroyl moieties were shown. Two aromatic proton signals at δ _H 5.98 (s) and 6.17 (s) with 6.71 (d, J=7.2 Hz) and 7.82 (d, J=7.2 Hz) confirmed camphorol aglycone. Additionally, β-binding of the glucosyl moiety was deduced from the coupling constant (J = 7.8 Hz) of the anomeric proton signal at δ _H 5.61 (Fig. 2(c)). The ¹³ C NMR spectrum of compound (16) showed a 41 carbon signal including 16 quaternary, 23 methine, 1 methylene and 1 methyl group (FIG. 2(c)).

NMR data analysis of Compound (16) was performed except for the addition of an acetyl group. Camphorol-3-O-[2",6"-di-OEp-coumaroyl]-β-D-glucopyranoside (Kaempferol -3-O-[2",6"-di-OEp-coumaroyl]-β-D-glucopyranoside). The sugar moiety linked to C-3 is H-1" (δ _H 5.61) and C-3 (δ _C 134.6) was confirmed by the HMBC correlation. H-3" (δ _H 5.12) and the HMBC correlation between the carbonyl group (δ _C 172.2) suggested the position of the acetyl group linked to C-3". Moreover, the bonding of the Ep-coumaroyl group was H-2" (δ _H 5.05) and C It was determined by the HMBC correlation between -γ"' (δ _C 168.1), H-6" (δH 4.15 and 4.27) and C-γ "" (δ _C 168.8) (FIG. 3 ). Therefore, the structure of compound (16) is camphorol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (Kaempferol-3-O -[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside).

virus compound CC50 ^a IC50 ^b TI ^c HRV1B Compound (4) >50 6.32±0.46 >7.91 Compound (5) >50 5.58±0.24 >8.96 Compound (8) >50 19.4±3.63 >2.57 Compound (9) >50 34.1±3.34 >1.46 Compound (12) 28.37 6.14±0.06 4.62 Compound (13) >50 1.22±0.01 >40.9 Compound (14) 28.19 5.89±0.21 4.78 Compound (15) >50 7.46±0.52 >6.70 Compound (16) >50 5.51±0.31 >9.07 Rupintrivir >50 <0.04 ND CVB3 Compound (14) >50 43.6±1.97 >1.14 Rupintrivir >50 7.88±0.94 >6.34 PR8 Compound (13) >50 37.1±3.62 >7.91 Compound (14) >50 28.2±0.69 >8.96 Oseltamivir >50 2.19±0.55 >22.8

* Results are expressed as mean IC ₅₀ values ± SD obtained from independent experiments performed three times.

^a Concentration required to reduce cell growth by 50% (μM).

^b The concentration required to inhibit 50% (μM) virus-induced CPE.

^c Therapeutic index = CC ₅₀ / IC ₅₀ .

To investigate effective antiviral phytochemicals, all isolated compounds were evaluated for antiviral activity by employing the SRB method using cytopathic effect (CPE) in HeLa or Vero cells. Rupintrivir against both HRV1B and CVB3 and Oseltamivir against PR8 were used as positive controls. Among the isolated compounds, most camphorol derivatives showed statistically significant antiviral activity against HRV1B-infected cells (Table 1). Among the compounds tested, camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside is against HRV1B-, CVB3- and PR8-infected cells. It showed the most consistent and effective antiviral activity.

conclusion

The phytochemical study of chestnut (C. crenata) chestnut was conducted with five cycloartane triterpene compounds of castartancrenoic acid AE and camphorol-3-O-[3"-acetyl-2",6"-di- One flavonoid compound of Ep-coumaroyl]-β-D-glucopyranoside and eleven additional compounds were isolated. To confirm antiviral phytochemicals, HRV1B-, CVB3- and PR8-infected Vero Or the isolated compounds for their biological activity against HeLa cells Antiviral assays showed that compounds derived from chestnut ( C. crenata ) were effective to treat viral infections caused by HRV1B, CVB3 and PR8. It has been demonstrated that it may be a viral agent.

The above description of the present invention is for illustrative purposes only, and those of ordinary skill in the art to which the present invention pertains will be able to understand that it can be easily modified into 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 limiting.

Claims (10)

An antiviral pharmaceutical composition containing one or more compounds selected from the following i) and ii) or a pharmaceutically acceptable salt thereof as an active ingredient:
I) Camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OZp -coumaroyl]-β-D-glucopyranoside), astragarin, tiliroside, cis-tiliroside, camphorol-3-O-[2"-OEp-cumaro Yl]-β-D-glucopyranoside (kaempferol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3-O-[3",6"-di- O-Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3- O-[2",6"-di-O-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OEp-coumaroyl]-β -D-glucopyranoside), camperol-3-O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[ 4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) and camphorol-3-O-[3"-acetyl-2",6"-di-Ep-coumaro One or more selected from the group consisting of]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) Flavonoid-based compounds comprising a; And
Ii) A cycloartan-based compound containing castaartancrenoic acid D or castaartancrenoic acid E.
The method of claim 1,
The compound is characterized in that isolated from the chestnut extract, antiviral pharmaceutical composition.
The method of claim 1,
The composition is characterized in that for combating a virus that causes respiratory diseases, antiviral pharmaceutical composition.
The method of claim 3,
The virus is characterized in that the Picornavirus family (Picornaviridae) or Orthomyxoviridae (Orthomyxoviridae), characterized in that, antiviral pharmaceutical composition.
The method of claim 4,
The Picornaviridae includes human rhinovirus 1B (HRV1B) or coxsackie virus B3 (CVB3),
The Orthomyxoviridae (Orthomyxoviridae) is characterized in that it comprises an influenza virus (PR8), antiviral pharmaceutical composition.
Antiviral health functional food containing one or more compounds selected from the following i) and ii) or a pharmaceutically acceptable salt thereof as an active ingredient:
I) Camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OZp -coumaroyl]-β-D-glucopyranoside), astragarin, tiliroside, cis-tiliroside, camphorol-3-O-[2"-OEp-cumaro Yl]-β-D-glucopyranoside (kaempferol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3-O-[3",6"-di- O-Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3- O-[2",6"-di-O-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OEp-coumaroyl]-β -D-glucopyranoside), camperol-3-O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[ 4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) and camphorol-3-O-[3"-acetyl-2",6"-di-Ep-coumaro One or more selected from the group consisting of]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) Flavonoid-based compounds comprising a; And
Ii) A cycloartan-based compound containing castaartancrenoic acid D or castaartancrenoic acid E.
An antiviral skin external preparation containing as an active ingredient at least one compound selected from the following i) and ii) or a pharmaceutically acceptable salt thereof:
I) Camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OZp -coumaroyl]-β-D-glucopyranoside), astragarin, tiliroside, cis-tiliroside, camphorol-3-O-[2"-OEp-cumaro Yl]-β-D-glucopyranoside (kaempferol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3-O-[3",6"-di- O-Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3- O-[2",6"-di-O-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OEp-coumaroyl]-β -D-glucopyranoside), camperol-3-O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[ 4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) and camphorol-3-O-[3"-acetyl-2",6"-di-Ep-coumaro One or more selected from the group consisting of]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) Flavonoid-based compounds comprising a; And
Ii) A cycloartan-based compound containing castaartancrenoic acid D or castaartancrenoic acid E.
An antiviral quasi-drug containing one or more compounds selected from the following i) and ii) or a pharmaceutically acceptable salt thereof as an active ingredient:
I) Camphorol-3-O-[2",6"-di-OZp-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OZp -coumaroyl]-β-D-glucopyranoside), astragarin, tiliroside, cis-tiliroside, camphorol-3-O-[2"-OEp-cumaro Yl]-β-D-glucopyranoside (kaempferol-3-O-[2"-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3-O-[3",6"-di- O-Ep-Coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[3",6"-di-OEp-coumaroyl]-β-D-glucopyranoside), camphorol-3- O-[2",6"-di-O-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[2",6"-di-OEp-coumaroyl]-β -D-glucopyranoside), camperol-3-O-[4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside (kaempferol-3-O-[ 4"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) and camphorol-3-O-[3"-acetyl-2",6"-di-Ep-coumaro One or more selected from the group consisting of]-β-D-glucopyranoside (kaempferol-3-O-[3"-acetyl-2",6"-di-Ep-coumaroyl]-β-D-glucopyranoside) Flavonoid-based compounds comprising a; And
Ii) A cycloartan-based compound containing castaartancrenoic acid D or castaartancrenoic acid E.
New compound represented by the following formula (1):
[Formula 1]

.
The method of claim 9,
The compound is characterized in that isolated from the chestnut extract, a novel compound.

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