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WO2022031151A1 - Composition antivirale comprenant un extrait de fougère aigle orientale ou une fraction de celle-ci - Google Patents

Composition antivirale comprenant un extrait de fougère aigle orientale ou une fraction de celle-ci Download PDF

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Publication number
WO2022031151A1
WO2022031151A1 PCT/KR2021/010527 KR2021010527W WO2022031151A1 WO 2022031151 A1 WO2022031151 A1 WO 2022031151A1 KR 2021010527 W KR2021010527 W KR 2021010527W WO 2022031151 A1 WO2022031151 A1 WO 2022031151A1
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WIPO (PCT)
Prior art keywords
extract
bracken
fraction
organic solvent
composition
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2021/010527
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English (en)
Korean (ko)
Inventor
박길홍
김태우
김명환
파티바다트리베니
헤마선다르알라빌리
순드라라잔나가순다라판디안
홍성수
최윤혁
최춘환
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ghpharm Co Ltd
Gyeonggido Business & Science Accelerator
Korea University Research and Business Foundation
Original Assignee
Ghpharm Co Ltd
Gyeonggido Business & Science Accelerator
Korea University Research and Business Foundation
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Application filed by Ghpharm Co Ltd, Gyeonggido Business & Science Accelerator, Korea University Research and Business Foundation filed Critical Ghpharm Co Ltd
Publication of WO2022031151A1 publication Critical patent/WO2022031151A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/11—Pteridophyta or Filicophyta (ferns)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system

Definitions

  • the present invention relates to an antiviral composition comprising a bracken extract or a fraction thereof, and more particularly, to a composition for preventing or treating RNA virus infection comprising a bracken rhizome extract or a fraction thereof as an active ingredient.
  • Virus means a toxic substance in Latin, and is a group of infectious pathogenic particles that pass through a bacterial filter paper (0.22 ⁇ m). Viruses can be classified into bacteriophages, plant viruses, and animal viruses according to the type of host cell, and can be classified into DNA viruses and RNA viruses according to the types of nucleic acids. Recently, various viral diseases such as swine flu, AI, coronavirus and foot-and-mouth disease have caused great social problems, and accordingly, concerns about effective countermeasures for viral diseases are arousing great social interest.
  • SARS-CoV-2 identified as the cause of COVID-19, spread worldwide and was recently declared an epidemic by the World Health Organization (WHO).
  • WHO World Health Organization
  • the new coronavirus is reported to be highly contagious and pathogenic, causing severe pneumonia symptoms in infected patients.
  • the virus has been found to be a serious threat, especially to the elderly and/or people with underlying diseases such as cardiovascular disease, diabetes, chronic respiratory disease and cancer.
  • an agent capable of effectively preventing and treating a novel virus infection such as SARS-CoV-2 has not yet been completed.
  • the present inventors repeated intensive research to develop an antiviral agent derived from a natural product that can effectively prevent and treat RNA virus infection. It was found that it inhibits and completed the present invention.
  • RNA virus infection comprising an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof as an active ingredient.
  • RNA virus infection consisting of an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof.
  • RNA virus infection essentially consisting of an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof.
  • Another object of the present invention is to provide a food composition for preventing or improving RNA virus infection comprising an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof as an active ingredient.
  • another object of the present invention is to provide a food composition for preventing or improving RNA virus infection consisting of a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • another object of the present invention is to provide a food composition for preventing or improving RNA virus infection, which is essentially composed of a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • Another object of the present invention is to provide an antiviral composition for RNA viruses comprising an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof.
  • Another object of the present invention is to provide an antiviral composition for RNA viruses consisting of a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • Another object of the present invention is to provide an antiviral composition for RNA viruses essentially consisting of an extract or an organic solvent fraction thereof.
  • Another object of the present invention is to provide a use of a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof for preparing a preparation for treating RNA virus infection.
  • Another object of the present invention is to provide a method for treating RNA virus infection comprising administering to an individual in need thereof an effective amount of a composition comprising an extract of Pteridium aquilinum or an organic solvent fraction thereof.
  • Another object of the present invention is to provide a use of a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof for preparing an antiviral agent for RNA virus.
  • Another object of the present invention is to provide an antiviral method against RNA viruses, comprising administering to an individual in need thereof an effective amount of a composition comprising a Pteridium aquilinum extract or an organic solvent fraction thereof.
  • the present invention provides a pharmaceutical composition for preventing or treating RNA virus infection comprising an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof as an active ingredient.
  • the present invention provides a pharmaceutical composition for preventing or treating RNA virus infection consisting of a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • the present invention provides a pharmaceutical composition for preventing or treating RNA virus infection essentially consisting of a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • the present invention provides a food composition for preventing or improving RNA virus infection comprising an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof as an active ingredient.
  • the present invention provides a food composition for preventing or improving RNA virus infection consisting of a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • the present invention provides a food composition for preventing or improving RNA virus infection essentially consisting of an extract or an organic solvent fraction thereof.
  • the present invention provides an antiviral composition for RNA viruses comprising an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof.
  • the present invention provides a composition for antiviral against RNA virus comprising a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • the present invention provides an antiviral composition for RNA viruses, which is essentially composed of a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • the present invention provides a use of a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof for preparing a preparation for treating RNA virus infection.
  • the present invention provides a method of treating an RNA virus infection comprising administering to an individual in need thereof an effective amount of a composition comprising a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof.
  • the present invention provides the use of a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof for preparing an antiviral agent for RNA viruses.
  • the present invention provides an antiviral method for RNA viruses, comprising administering to an individual in need thereof an effective amount of a composition comprising a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof do.
  • the present invention provides a pharmaceutical composition for preventing or treating RNA virus infection, comprising an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof as an active ingredient.
  • the fern Pteridium aquilinum
  • the thick underground stem extends and leaves come out, and the young leaves are curled and covered with hairs like white cotton. Mature leaves are pinnate compound leaves. The young leaves are eaten as a vegetable.
  • the bracken extract is preferably a rhizome extract of bracken.
  • extract refers to an extract obtained by extracting bracken, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a prepared or purified product of the extract, or a mixture thereof, such as the extract itself and It includes extracts of all formulations that can be formed using the extract.
  • the method of extracting the bracken 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, ultrasonic extraction, filtration, reflux extraction, and the like, and these may be performed alone or in combination of two or more methods.
  • the bracken extract of the present invention may include a pretreatment process for the raw material in order to increase the extraction efficiency during the preparation of the extract, for example, may be used after drying the bracken and then chopped.
  • the extract of the present invention may be extracted using one or more selected from the group consisting of water and an organic solvent having 1 to 8 carbon atoms.
  • the extraction solvent may be edible single or multiple types of solvents, but preferably water, alcohol, and alcohols having 1 to 6 carbon atoms [methanol (C1), ethanol (C2), propanol (C3), butanol ( C4), pentanol (C5), hexanol (C6)] may be used at least one selected from the group consisting of. More preferably, in the present invention, the bracken extract may be water extracted with a solvent.
  • the extract of the present invention may be used in a liquid form by filtration and/or concentration, and may be used after being solidified through a conventional drying process such as spray drying or freeze drying.
  • a drying process such as spray drying or freeze drying.
  • dextrin or the like may be mixed and dried before spray drying or freeze drying.
  • fraction refers to a result obtained by performing fractionation in order to separate a specific component or a specific group of components from a mixture including various components.
  • the fractionation method for obtaining the fraction in the present invention is not particularly limited, and may be performed according to a method commonly used in the art.
  • Non-limiting examples of the fractionation method include a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (size, charge, hydrophobicity) or a chromatographic fractionation method for performing separation according to affinity), and combinations thereof.
  • a method of obtaining a fraction from the extract by treating the extract obtained by extracting the bracken rhizome with a predetermined solvent.
  • the type of the fractionation solvent used to obtain the fraction is not particularly limited, and any solvent known in the art may be used.
  • the fractionation solvent include water, a polar solvent such as an alcohol having 1 to 4 carbon atoms; hexane (Hexan), ethyl acetate (Ethyl acetate), chloroform (Chloroform), a non-polar solvent such as dichloromethane (Dichloromethane); or a mixed solvent thereof.
  • the fraction may be an ethyl acetate fraction or a butanol fraction of a bracken hot water extract.
  • extract or fraction may be prepared and used in the form of a dry powder, but is not limited thereto.
  • the virus may be an RNA virus.
  • RNA virus refers to a virus that uses reverse transcriptase to convert its own genetic information made of RNA into DNA and inserts it into the DNA of the host, thereby causing the host to replicate the genetic information of the virus instead.
  • the type of the RNA virus is not particularly limited, and Amalgaviridae, Birnaviridae, Chrysoviridae, Cystoviridae, Endornaviridae ( Endornaviridae), Hypoviridae, Megabirnaviridae, Partitiviridae, Picobirnaviridae, Reoviridae, Totiviridae, Quadriviridae, Arteriviridae, Coronaviridae, Mesoniviridae, Roniviridae, Dicistroviridae, Iflaviridae (Iflaviridae), Marnaviridae, Picornaviridae, Secoviridae, Alphaflexiviridae, Betaflexiviridae, Gammaflexiviridae ), Tymoviridae, Bornaviridae, Filoviridae, Paramyxoviridae, Rhabdoviridae, Nyamiviridae Caliciviridae, Fla
  • the RNA virus is an influenza virus, influenza A virus subtype H1N1, avian influenza virus, rhinovirus, coronavirus. ), parainfluenza virus, respiratory syncytial virus, human immunodeficiency virus (HIV), retrovirus and hepatitis C virus. have.
  • the coronavirus is selected from the group consisting of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome (MERS) coronavirus.
  • SARS-CoV-2 severe acute respiratory syndrome coronavirus 2
  • SARS-CoV severe acute respiratory syndrome coronavirus
  • MERS Middle East respiratory syndrome coronavirus
  • the pharmaceutical composition according to the present invention may be formulated in various ways according to the route of administration by methods known in the art together with a pharmaceutically acceptable carrier.
  • the carrier includes all kinds of solvents, dispersion media, oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads and microsomes.
  • the pharmaceutical composition according to the present invention may be administered to a patient in a pharmaceutically effective amount, that is, an amount sufficient to prevent or alleviate symptoms and treat RNA virus infection.
  • a typical daily dose may be administered in the range of about 0.01 to 1000 mg/kg, preferably, it may be administered in the range of about 1 to 100 mg/kg.
  • the pharmaceutical composition of the present invention may be administered once or divided into several doses within a preferred dosage range.
  • the dosage of the pharmaceutical composition according to the present invention may be appropriately selected by those skilled in the art according to the route of administration, administration, age, sex, body weight, individual differences, and disease state.
  • the route of administration may be oral or parenteral administration.
  • Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, rectal, or pancreatic administration. It may be administered internally, but is not limited thereto, and most preferably, it may be administered orally.
  • composition of the present invention When the pharmaceutical composition of the present invention is orally administered, powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers according to methods known in the art together with a suitable oral administration carrier It can be formulated in the form of, etc.
  • suitable carriers include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, and starches, including corn starch, wheat starch, rice starch and potato starch, cellulose, Cellulose, including methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethylcellulose, and the like, fillers such as gelatin, polyvinylpyrrolidone, and the like may be included. In addition, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant if necessary. Furthermore, the pharmaceutical composition may further include an anti-aggregating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, and a preservative.
  • sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylit
  • composition of the present invention when administered parenterally, may be formulated according to methods known in the art in the form of injections, transdermal administration agents, nasal inhalants, etc. together with suitable parenteral carriers.
  • the pharmaceutical composition may be administered by any device capable of transporting the active agent to a target cell.
  • Preferred administration methods and formulations are intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, or drip injections.
  • Injections include aqueous solvents such as physiological saline or Ringer's solution, vegetable oil, higher fatty acid esters (eg, ethyl oleate), and non-aqueous solvents such as alcohols (eg, ethanol, benzyl alcohol, propylene glycol or glycerin).
  • It can be prepared using, stabilizers for preventing deterioration (eg, ascorbic acid, sodium hydrogen sulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for pH adjustment, to inhibit the growth of microorganisms and a pharmaceutical carrier such as a preservative (eg, phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).
  • stabilizers for preventing deterioration eg, ascorbic acid, sodium hydrogen sulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.
  • emulsifiers eg., buffers for pH adjustment, to inhibit the growth of microorganisms and a pharmaceutical carrier such as a preservative (eg, phenylmercuric nitrate, thimerosal, benz
  • suitable carriers include, but are not limited to, water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof, and/or a solvent or dispersion medium containing vegetable oil.
  • suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with triethanolamine or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. etc. can be used.
  • the injection may further include various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
  • various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
  • the injection may further contain an isotonic agent such as sugar or sodium chloride.
  • the pharmaceutical composition according to the present invention may contain one or more buffers (eg saline or PBS), carbohydrates (eg glucose, mannose, sucrose or dextran), antioxidants, bacteriostats, chelating agents (eg EDTA or glutathione), adjuvants (eg, aluminum hydroxide), suspending agents, thickening agents and/or preservatives.
  • buffers eg saline or PBS
  • carbohydrates eg glucose, mannose, sucrose or dextran
  • antioxidants eg glucose, mannose, sucrose or dextran
  • bacteriostats eg EDTA or glutathione
  • adjuvants eg, aluminum hydroxide
  • suspending agents eg, thickening agents and/or preservatives.
  • compositions of the present invention may be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.
  • composition of the present invention may be administered alone or in combination with a known compound having a therapeutic effect on viral infection.
  • the present invention also provides a food composition for preventing or improving RNA virus infection, comprising an extract of bracken ( Pteridium aquilinum ) or an organic solvent fraction thereof as an active ingredient.
  • the food composition of the present invention includes all types of functional foods, nutritional supplements, health foods, and food additives.
  • Food compositions of this type can be prepared in various forms according to conventional methods known in the art.
  • the lactic acid bacteria fermented product of bracken extract can be prepared in the form of tea, juice, and drink and ingested by liquefaction, granulation, encapsulation and powdering so that it can be consumed.
  • it can be prepared in the form of a composition by mixing the bracken extract and a known active ingredient known to be effective against RNA virus infection.
  • functional foods include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods thereof (eg, canned fruit, bottled, jam, marmalade, etc.), fish, meat, and processed foods (eg, ham, sausages) corn beef, etc.), breads and noodles (eg udon noodles, soba noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, syrup, dairy products (eg butter, cheese, etc.), edible vegetable oils and fats, margarine, vegetable It can be prepared by adding the fermented product of bracken extract to protein, retort food, frozen food, various seasonings (eg, soybean paste, soy sauce, sauce, etc.). In addition, in order to use the bracken extract in the form of an additive, it may be prepared and used in the form of a powder or a concentrate.
  • beverages including alcoholic beverages
  • fruits and processed foods thereof eg, canned fruit, bottled, jam, marmalade, etc.
  • the preferred content of the bracken extract in the food composition of the present invention is not limited thereto, but is preferably 0.1 to 90% by weight of the final prepared food. More preferably, the food composition containing the bracken extract of the present invention as an active ingredient may be prepared in the form of a health functional food or dietary supplement by mixing with an active ingredient known to have a therapeutic effect on RNA virus infection. .
  • the present invention also provides a composition for antiviral against RNA virus comprising a bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof.
  • the antiviral composition includes any composition except for the pharmaceutical composition and food composition described above, and non-limiting examples thereof include an antiviral cosmetic composition, an antiviral external skin composition, and an antiviral quasi-drug composition. etc. may be included.
  • suitable formulations include, for example, a solution, a gel, a solid or kneaded anhydrous product, an emulsion obtained by dispersing an oil phase in an aqueous phase, a suspension, a microemulsion, microcapsules, microgranules, or It may be provided in the form of an ionic (liposome), non-ionic vesicular dispersion, cream, skin, lotion, powder, ointment, or spray. In addition, it may be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.
  • the cosmetic composition or composition for external application to the skin may include, in addition to the compound, a fatty substance, an organic solvent, a solubilizer, 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 nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or external preparations for the skin. It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.
  • the composition is a quasi-drug composition
  • the bracken ( Pteridium aquilinum ) extract or an organic solvent fraction thereof may be added as it is, or it may be used together with other quasi-drugs or quasi-drug components, and may be appropriately mixed according to a conventional method.
  • the mixing amount of the active ingredient may be suitably determined according to the purpose of use.
  • the type of the quasi-drug is not particularly limited, but, for example, it may be a disinfectant cleaner, shower foam, gargrin, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment, patch, or filter filler.
  • the antiviral composition provided by the present invention can be utilized in daily life to prepare for RNA virus infection.
  • the present invention provides a use of a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof for preparing a preparation for treating RNA virus infection.
  • the present invention provides a method for treating an RNA virus infection comprising administering to an individual in need thereof an effective amount of a composition comprising a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof.
  • the present invention provides a use of a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof for preparing an antiviral agent for RNA virus.
  • the present invention provides an antiviral method against RNA viruses, comprising administering to an individual in need thereof an effective amount of a composition comprising a bracken (Pteridium aquilinum) extract or an organic solvent fraction thereof.
  • the 'effective amount' of the present invention refers to an amount that exhibits the effect of improving, treating, preventing, detecting, diagnosing, or inhibiting or reducing RNA virus infection when administered to an individual
  • the "individual" refers to an animal, preferably Preferably, it may be an animal, including a mammal, particularly a human, and may be an animal-derived cell, tissue, organ, or the like. The subject may be a patient in need of the effect.
  • the 'treatment' of the present invention refers generically to alleviating the symptoms of RNA virus infection or RNA virus infection, which may include curing, substantially preventing, or ameliorating the condition of the disease. including, but not limited to, alleviating, curing or preventing one or most symptoms resulting from the disease.
  • the term “comprising” is used synonymously with “including” or “characterized by”, and in a composition or method according to the present invention, specifically Additional components or method steps that have not been excluded are not excluded. Also, the term “consisting of” means excluding additional elements, steps, or components not specifically described. The term “essentially consisting of” means that, in the scope of a composition or method, it may include substances or steps that do not substantially affect its basic properties in addition to the substances or steps described.
  • the bracken extract provided by the present invention has very excellent activity of inhibiting reverse transcriptase and inhibiting the proliferation of RNA virus, and thus can be very usefully utilized for preventing or developing a therapeutic agent for RNA virus infection.
  • MMLV Moloney murine leukemia virus reverse transcriptase inhibitory activity of bracken rhizome extracts or fractions (EA: ethyl acetate, Bu: butanol).
  • AMV Alfa myeloblastosis virus
  • EA ethyl acetate
  • Bu butanol
  • FIG. 3 is a result of evaluating the antiviral activity of a bracken rhizome hot water extract (A), an ethyl acetate fraction thereof (B), and a fraction (C) excluding the ethyl acetate fraction (C) in SARS-CoV-2 infected vero cells.
  • bracken hot water extract After boiling the bracken rhizome in water at 100° C. for 48 hours, the solid component was removed to obtain a bracken hot water extract.
  • the bracken hot water extract was mixed with the same volume of ethyl acetate, left for 1 hour, and then centrifuged to separate the supernatant hot water layer and ethyl acetate layer, and each was used as a sample. Ethyl acetate was evaporated from the obtained bracken ethyl acetate extract using a rotary evaporator. The ethyl acetate fraction of the bracken hot water extract thus obtained was dissolved in a minimum amount of dimethylsulfoxide (DMSO) and completely dissolved with phosphate-buffered saline (PBS) to be used as the bracken ethyl acetate fraction.
  • DMSO dimethylsulfoxide
  • PBS phosphate-buffered saline
  • bracken extract 5 kg of bracken rhizomes purchased in Namwon-si, Jeollanam-do were washed clean, and 30L of water was added, boiled for 48 hours, and then stored in a refrigerator was used as a hot water extract.
  • the EA fraction was dissolved in a minimum amount of DMSO and then diluted with PBS. At this time, the dilution factor was diluted to 70% of the original volume of the hot-water extract in anticipation of a yield of approximately 70%.
  • the hot water extract was added with the same volume of butanol (Bu), mixed well, centrifuged to separate and collected the hot water layer and the butanol layer of the supernatant, and then the butanol layer was dried with a rotary evaporator to form a Bu fraction.
  • Bu fraction was dissolved in a minimum amount of DMSO and diluted with water. At this time, the dilution factor was diluted to 70% of the original volume of the hot-water extract, expecting a yield of approximately 70%.
  • C6 rat glial cell
  • NIH3T3 mouse embryo fibroblast cell-line
  • LD 50 Lethal Dose 50
  • MMLV Moloney murine leukemia virus
  • AMV Avian myeloblastosis virus
  • RNA virus TaKaRa, Japan
  • reverse transcription reaction solution 5X reverse transcription buffer, RNase inhibitor, MMLV or AMV reverse transcriptase, DEPC-treated water, and reverse transcriptase inhibitory material were mixed well in a total volume of 10 ⁇ L to prepare a reverse transcription reaction solution.
  • candidates for reverse transcriptase inhibition bracken hot water extract, EA fraction, EA supernatant hot water layer, Bu fraction, and Bu supernatant hot water layer were used.
  • the reverse transcription reaction solution was added to the priming reaction solution and mixed well.
  • the reverse transcription reaction was carried out at 45° C. for 30 minutes. After terminating the reaction by treatment at 85 °C for 5 minutes, cDNA was precipitated with ice-cold ethanol, and cDNA was obtained by centrifugation and stored at -20 ° C until Taq PCR reaction.
  • the cDNA synthesized in this way was amplified by Taq PCR (Elpisbio) to confirm cDNA synthesis ( FIGS. 1 and 2 ).
  • PCR grade distilled water, Taq polymerase, buffer, primer, and cDNA were mixed so that the volume of the total reaction solution was 20 ⁇ L.
  • the reaction solution was subjected to 35 cycles of initial denature 95 °C 3 min, denature 95 °C 10 sec, anneal Tm-4 °C 10 sec, extend 72 °C 10 sec/kbp.
  • the amplified DNA was separated by agarose gel electrophoresis and visually confirmed.
  • Vero cells were cultured in DMEM (Dulbecco's modified Eagle's medium; Gibco Lab Inc., Grand Island, NY) containing 10% FBS at 37°C and 5% CO 2 .
  • Vero cells cultured in 96 wells cell plate (Greiner, Nurtingen, Germany) were dispensed at 1 x 10 4 cells per well and cultured for 24 hours.
  • Betacov/Korea/KCDC03/2020 virus diluted to 100TCID 50 /50 ⁇ L concentration was infected in Vero cells cultured in 96 wells cell plate. Incubated for 1 hour and 30 minutes at 37° C. and a concentration of 5% CO 2 .
  • the prepared bracken hot water extract (A), EA fraction (B), and fraction (C) excluding the EA fraction (C) were diluted 2-fold and 10-fold stepwise, and No FBS DMEM containing TPCK treated trypsin at a concentration of 0.2 ⁇ g/mL replaced with Incubated for 72 hours at a temperature of 37° C. and a concentration of 5% CO 2 .
  • SARS-CoV-2 virus inhibited by bracken extract was identified for CPE and stained with 10% Crystal Violet (Sigma-aldrich, St. Louis, MO) to determine the minimum antiviral inhibitory concentration. Remdesivir (RDV) was used as a positive control.
  • the bracken extract provided by the present invention has very excellent activity of inhibiting reverse transcriptase and inhibiting the proliferation of RNA virus, and thus can be very usefully utilized for preventing or developing a therapeutic agent for RNA virus infection.

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Abstract

La présente invention concerne une composition antivirale comprenant un extrait de fougère aigle orientale ou une fraction de celle-ci et, plus particulièrement, une composition comprenant un extrait de racine de fougère aigle orientale ou une fraction de celle-ci en tant que principe actif pour la prévention ou le traitement de symptômes d'infection virale à ARN. L'extrait de fougère aigle orientale selon la présente invention a une excellente activité inhibitrice vis-à-vis de la transcriptase inverse et de la prolifération virale à ARN et en tant que tel, peut être très avantageusement utilisé pour développer un agent prophylactique ou thérapeutique contre des symptômes d'infection virale à ARN.
PCT/KR2021/010527 2020-08-07 2021-08-09 Composition antivirale comprenant un extrait de fougère aigle orientale ou une fraction de celle-ci Ceased WO2022031151A1 (fr)

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KR102532946B1 (ko) 2022-04-20 2023-05-17 한국 한의학 연구원 복합한약재 추출물을 유효성분으로 포함하는 코로나바이러스 감염증의 예방, 개선 또는 치료용 조성물

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Publication number Priority date Publication date Assignee Title
WO2023212353A1 (fr) * 2022-04-29 2023-11-02 Emory University Inhibiteurs de l'entrée de virus dérivés de plantes

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