WO2012144754A2 - Composition contenant un extrait de galla rhois présentant des effets inhibiteur de virus, ou un composé isolé à partir de celui-ci, comme ingrédient actif et ses utilisations - Google Patents

Composition contenant un extrait de galla rhois présentant des effets inhibiteur de virus, ou un composé isolé à partir de celui-ci, comme ingrédient actif et ses utilisations Download PDF

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WO2012144754A2
WO2012144754A2 PCT/KR2012/002450 KR2012002450W WO2012144754A2 WO 2012144754 A2 WO2012144754 A2 WO 2012144754A2 KR 2012002450 W KR2012002450 W KR 2012002450W WO 2012144754 A2 WO2012144754 A2 WO 2012144754A2
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ethyl
virus
gallate
extract
influenza virus
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PCT/KR2012/002450
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WO2012144754A3 (fr
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박현
김경민
송현옥
김성연
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원광대학교 산학협력단
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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
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/22—Anacardiaceae (Sumac family), e.g. smoketree, sumac or poison oak
    • 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
    • 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
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses

Definitions

  • the present invention relates to a composition and its use containing a gall extract or a compound isolated therefrom as an active ingredient having a virus inhibitory effect.
  • Influenza virus is an RNA virus belonging to the family Orthomyxoviridae, and serotypes are classified into three types, type A, type B and type C. Among them, hepatitis B and C have been identified only in humans, while hepatitis A has been identified in humans, horses, pigs, other mammals and various types of poultry and wild birds (Selmons et al., Avian Dis. , 18 (1), pp 119-124, 1974; Webster RG et al., Microbiol Rev., 56 (1), pp 152-179, 1992).
  • the serotypes of influenza A virus are classified according to the two types of proteins on the surface of the virus: hemagglutinin (HA) and neuraminidase (NA), and 144 types (HA protein) 16 species and 9 NA proteins).
  • HA acts as a virus to attach to somatic cells
  • NA allows the virus to germinate from infected cells and penetrate into other cells (Alexander DJ, Vet. Microbiol., 74 (1-2), pp3-13, 2000 ).
  • influenza A virus The normal natural host of influenza A virus is known as wild waterfowl such as ducks, seagulls, etc.As a result of epidemiological investigations of influenza virus infection in wild birds around the world, all 16 existing HA and 9 NA types Influenza viruses have been confirmed to infect wild birds (Selmons et al., Avian Dis., 18 (1), pp119-124, 1974).
  • Avian influenza virus, classified as type A is a common infectious disease virus, a non-pathogenic avian influenza virus that causes mild respiratory symptoms according to pathogenicity when infected with chickens, and causes mortality and spawning degradation of about 1-30%.
  • LPAI Low pathogenic avian influenza
  • HPAI high pathogenic avian influenza
  • OIE International Water Services Bureau
  • the first record of human influenza dates back to 412 B.C.E., but the first pandemic influenza record of civilization is estimated to have occurred throughout Europe from 1173 to 1174 (Grmek MD, Les Maladies a L '). aube de la Civilization Accidentale, Payot, Paris, 1893). Since the 20th century, the record of influenza has been more scientifically treated and based on data left, there have been three pandemic influenza virus outbreaks in humans since the 20th century. The first pandemic of the influenza pandemic (aka Spanish flu) since the 20th century, which was in pandemic between 1918 and 1920, has been recorded as the greatest damage to civilization, and between 20 and 50 million people died during that period. (Walter JH, Bull. NY Acad.
  • the Spanish influenza virus is still a major epidemic of pandemic flu, which is still spreading worldwide every year, and has been identified as A / H1N1, a serotype very similar to the swine flu virus, but the RNA polymerase (PAymerase) PA, PB1 of Spanish influenza virus.
  • the PB2 subunit is derived from algae and has been reported to be infected by humans during the adaptation of the virus (Taubenberger JK et al., Nature, 437, pp889-893, 2005). ).
  • the second pandemic of the influenza virus since the 20th century, which occurred between 1957 and 1958, began in China and rapidly spread along the coast to nearby Hong Kong, Singapore, Japan and Taiwan in six to seven months.
  • the causative virus is serotype A / H3N2, which differs from the previous serotype H2N2 virus and HA, which has been analyzed to be transmitted from birds, and is still the main epidemic of pandemic flu, which is still prevalent worldwide every year. (Oxford JS, Rev. Med. Virol., 10 (2), pp 119-133, 2000).
  • serotypes of avian influenza virus are infected by humans, causing flu and causing death.
  • avian-induced avian influenza viruses including serotype A / H5N1, which has been called “Hong Kong bird flu" to date, and serotype A / H7N7, and serotype A / H9N2, can infect humans. It is estimated that there is a possibility. Therefore, the present study on the mutant viruses derived from these three serotypes is being conducted worldwide (Suarez DL et al., J. Virol., 72 (8), pp6678-6688, 1998).
  • the mutant virus is an avian influenza virus belonging to serotype A / H5N1, which is also avian influenza virus that has been prevalent only in birds for the first time among humans.
  • Influenza viruses can infect the respiratory tract, cause systemic symptoms, change their appearance periodically, and move to other hosts without killing them before they die. Although it is the virus that causes the greatest economic loss to human beings and preventive vaccines have been developed, they have not caught up with the mutation of the virus, and there is no fundamental virus treatment yet.
  • Amantadine and rimantadine are substances that inhibit the function of M2 ion channel proteins of influenza viruses and are representative antiviral agents that inhibit the growth of influenza viruses in vivo. However, these two antiviral agents were found to be effective only against serotype A influenza virus and not against serotype B influenza virus without M2 protein. In addition, amantadine and rimantadine have been found to have the disadvantage that the emergence of a mutant virus that does not affect the ion channel function of influenza virus M2 protein when used.
  • influenza virus Since the influenza virus was resistant to amantadine and rimantadine during the flu season of 2005-2006, CDC recommended discontinuing amantadine and rimantadine at this time (MMWR Morb Mortal Wkly Rep., 55, pp 44-46). , 2006).
  • Zanamivir and oseltamivir which are developed to compensate for these drawbacks, inhibit the function of the neuraminidase protein of influenza viruses and are representative of the suppression of influenza virus proliferation in vivo.
  • Antiviral agents These two antiviral agents are known to be effective against all 16 serotype A influenza virus viruses and serotype B influenza virus viruses.
  • zanamivir has the disadvantage of inhalation and intravenous administration, while oseltamivir can be administered orally, but it has been pointed out as a disadvantage due to side effects such as recent reports of the emergence of resistant virus and vomiting and dizziness upon oral administration (Ward P et. al., J. antimicrob. Chemother., 55 (supp1), pp5-21, 2005). Therefore, the need for the development of new anti-influenza preparations continues to emerge.
  • Coronaviruses are single-stranded RNA viruses of 100-160 nm in diameter in various forms. Its name derives from what appears to be a crown because of the club-shaped spine that is expressed from the viral envelope. Coronaviruses are divided into three antigen groups.
  • the representative animal-type coronaviruses include MHV (serum group II), a species that infects the gastrointestinal, liver and nervous systems in rodents such as rats and causes pathologies.
  • MHV-JHM and MHV-A59 strains were studied the most because the infection of the nervous system causes encephalomyelitis and cerebral infarction. This pathology is very similar to human pathology.
  • HCoV-229E and HCoV-OC43 coronavirus strains which cause colds in humans, are human isolates belonging to serogroups I and II, respectively, and the nasopharyngeal cilia through the aminopeptidase N receptor or sialic acid receptor. Infect epithelial cells. Recently, coronaviruses HCoV-NL63 (Group I) and HCoV-HKUI (Group II) were further isolated from patients admitted for acute respiratory disease (van der Hoek et al., 2004 Nature Med., 10, pp68-373, 2004; Woo et al., J.
  • SARS coronavirus-related diseases known as SARS between 2002 and 2003. The epidemic, which began in southern China, spread to 28 countries in Asia, Europe and the Americas, with about 8,000 patients. These natural pathogens of SARS-CoV are presumed to be Horseshoe bats, and are believed to have begun when humans come into contact with infected animals such as the Palm civet. SARS coronavirus is a virus that can cause a common infectious disease.
  • SARS coronavirus infects the human respiratory tract through the angiotensin convertase 2 receptor, the propagation mechanism of which is not fully understood and there is no proven treatment.
  • Ribavirin has been frequently used but has little in vitro effect on SARS-CoV and no effect on disease progression (Fauci et al., Harrison's Principle of Internal Medicine, 17th Ed., Vol 1, pp1362-1364, 2009; Knipe DM and Howley PM, Fields Virology, 5th Ed., Vol 1, pp 1307-1323, 2007).
  • anti-coronavirus preparations for human-infected coronaviruses are required as well as anti-influenza preparations.
  • the present inventors considered natural ingredients as a novel substance which is completely different from existing compounds, is less toxic, and can be a new target in the current situation in which the necessity of the development of new anti-influenza and anti-coronavirus preparations is continuously required.
  • the development using natural ingredients is more likely to succeed at a lower cost than the development of organic compound-derived formulations using chemical compound discovery. It is economical.
  • natural products are preferred for the prevention of early diseases and infectious diseases, the market is growing rapidly and is expected to replace side effects and resistance problems caused by long-term administration of existing antiviral agents.
  • the gall bladder is a worm house in which aphid aphid ( Aphis chinensis J. Bell) is stabbed on the leaves of Rhus javanica L. belonging to the family Anacardiaceae.
  • the outer surface is grayish brown with light hairs, 3 ⁇ 7cm long, 2 ⁇ 5cm wide and 2mm thick, and hard and easily broken.
  • the inside is usually empty or grayish white with dead bugs and secretions, sometimes with a disgusting smell.
  • the harvested gall are used by drying, boiling or steaming in the sun. Its main ingredient is 50 ⁇ 70% of pyrogallol tannin, and it contains malic acid and gallic acid. It is used as astringent, branch, and hemostatic agent for diarrhea, sputum, diabetes, bleeding, and anemia (Information, et al.
  • the gall bladder extract or the compound isolated therefrom exhibited anti-influenza effect and the mechanism of action thereof.
  • the anti-germ extract and the compound isolated therefrom were subjected to antiviral experiments against influenza virus, avian influenza virus, and animal-infected coronavirus, confirming their excellent antiviral effect to complete the present invention.
  • the present invention is a new influenza virus, algae containing as an active ingredient Galla Rhois extract (TCM-33) or a compound isolated therefrom having low toxicity and resistance to living organisms and exhibiting antimicrobial activity.
  • TCM-33 Galla Rhois extract
  • the present invention is an RNA polymerase of a new influenza virus, avian influenza virus and human infectious or acquired common infectious coronavirus containing Galla Rhois extract (TCM-33) or a compound isolated therefrom as an active ingredient. (polymerase) and neuraminidase inhibitors are provided.
  • Seasonal flu or swine flu viruses as defined herein are A / H1N1, A / H2N2, A / H3N1, A / H3N2, A / H3N8 and the like and avian influenza viruses are A / H9N2 (A / Chicken / Korea / MS96 / 1996) , A / H1N1, A / H5N1, A / H7N7, A / H7N2, A / H7N3, etc.
  • the human infection-type coronavirus is HCoV-229E (Group I), HCoV-NL63 (Group II), HCoV-OC43 (Group II), HCoV-HKUI (Group II) And SARS-CoV (Group II).
  • Other animal-infected coronaviruses include MHV-A59, MHV-JHM, and stomatitis virus includes VSV.
  • influenza virus and human infectious coronavirus Diseases caused by influenza virus and human infectious coronavirus as defined herein include seasonal flu, swine flu, avian influenza, common cold or acute respiratory syndrome (SARS).
  • SARS common cold or acute respiratory syndrome
  • the disease is characterized by occurring in mammals including humans.
  • composition containing the Galla Rhois extract (TCM-33) or a compound separated therefrom of the present invention as an active ingredient comprises 0.1 to 50% by weight of the extract or a compound separated therefrom based on the total weight of the composition. .
  • gallic acid (1) paradigal acid and metadigal acid represented by the following structural formulas (1) to (5) A mixture of methyl p -digallic acid and methyl m -digallic acid (2), penta- O -galloyl glucose (3), ethyl gallate (4) or ethyl para A mixture of ethyl p -digallate and ethyl m -digallate; 5, preferably a mixture of p -digallic acid and m -digallic acid) or ethyl p-di-gallate and ethyl methacrylate complex of di-gallate (a mixture of ethyl p -digallate and ethyl -digallate m), more preferably those of 1 to 2: 1 by weight of the composite, and most preferably Preferably, a new influenza containing a 1:
  • the extract is a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water, ethanol or a mixed solvent thereof, more preferably a 50 to 90% ethanol mixed solvent, even more preferred.
  • a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water, ethanol or a mixed solvent thereof, more preferably a 50 to 90% ethanol mixed solvent, even more preferred.
  • the gall bladder extract of the present invention may be dried by cutting dried gall bladder into about 1 to 30 times water, preferably about 10 to 20 times dry weight, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably water. , Ethanol or a mixed solvent thereof, more preferably a 50 to 90% ethanol mixed solvent, even more preferably a 70 to 80% ethanol mixed solvent, at an extraction temperature of 20 to 100 ° C., preferably 80 to 100 ° C.
  • extraction method such as cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, preferably 1 to 5 times obtained using a reflux cooling extraction method, Preferably repeated extraction 2 to 3 times and then concentrated under reduced pressure to obtain the five gall extract of the present invention.
  • the present invention is obtained from the above production method and the above production method, gallic acid (1) as described in Fig. 1 (HPLC) herein, a complex of paradigal acid and metadigal acid (2), ethyl gallate (4) and a quinine 70% ethanol extract (hereinafter referred to as GR70E) consisting of a content ratio (4.3: 1: 7.2: 16.3) of the complex of ethyl paradigalate and ethyl metadigallate (5).
  • GR70E quinine 70% ethanol extract
  • the first step to obtain the gallastula extract A second step of dissolving the crude extract in distilled water and then eluting with distilled water, 50% aqueous ethanol and ethanol on an Amberlite XAD-2 resin column; A third step of subjecting the 50% aqueous ethanol eluting fraction obtained in the second step to chromatography and dividing into seven fractions; A fourth step of dividing the first fraction from the third fraction into three fractions by chromatography; A fifth step of separating the first fraction from the fourth step fraction by silica gel column chromatography to obtain gallic acid; A sixth step of subjecting the second fraction of the fourth step fraction to column chromatography to obtain a complex of paradigal acid and metadigal acid; A seventh step of subjecting the third fraction of the fourth fraction to column chromatography to obtain pentao-galloyl glucose; An eighth step of column chromatography of the fourth fraction of the fractions of the fourth step to obtain ethyl gallate; The sixth fraction of the fraction of the fraction of the fractions
  • the present invention relates to a novel influenza virus, avian influenza virus, and human infectious or acquired common infection-type coronavirus containing a Galla Rhois extract or a compound isolated therefrom as an active ingredient.
  • the gall bladder extract of the present invention and the compounds isolated therefrom have low toxicity and resistance, and also conducted antiviral experiments against swine flu virus, avian influenza virus, and animal infected corona virus, and confirmed their excellent antiviral effect. It was. Molecular modeling studies have also shown that compounds isolated from gall bladder extracts may exhibit intense inhibitory activity by acting as target molecules on RNA polymerase and neuraminidase of the H1N1 influenza virus, H1N1, avian influenza virus and human infectious agents. In addition, it has been found to be useful for the treatment and prevention of seasonal influenza, swine flu, avian influenza, common cold or severe respiratory syndrome (SARS) caused by acquired common infectious coronavirus.
  • SARS severe respiratory syndrome
  • the pharmaceutical composition of the present invention comprises 0.1 to 50% by weight of the extract or compound based on the total weight of the composition.
  • compositions containing extracts or compounds of the invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions containing extracts or compounds of the invention.
  • compositions or compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.
  • compositions containing extracts or compounds according to the invention are in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used.
  • Carriers, excipients and diluents which may be contained in the composition containing the compound include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
  • diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. ) Or lactose, gelatin and the like are mixed.
  • lubricants such as magnesium stearate and talc are also used.
  • Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin.
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
  • the non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.
  • As the base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
  • Preferred dosages of the extracts or compounds of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art.
  • the extract or compound of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 10 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
  • the extracts or compounds of the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
  • the present invention is for the prevention and improvement of diseases caused by the new influenza virus, avian influenza virus and human infectious or acquired common infection type coronavirus containing Galla Rhois extract or a compound isolated therefrom as an active ingredient.
  • Examples of the food to which the extract or compound of the present invention can be added include various foods, beverages, gums, teas, vitamin complexes, and health functional foods.
  • the amount of the extract or compound in the food or beverage may be added at 0.01 to 15% by weight of the total food weight, the health beverage composition is 0.02 to 5 g, preferably 0.3 to 1g based on 100 ml Can be added.
  • Health functional food of the present invention includes the form of tablets, capsules, pills, liquids and the like.
  • the health functional beverage composition of the present invention is not particularly limited to other ingredients except for containing the extract or compound as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks.
  • natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • natural flavoring agents such as, tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.
  • the proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.
  • the extracts or compounds of the present invention are various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like.
  • the extracts or compounds of the present invention may contain fruit flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the extract or compound of the present invention.
  • the present invention includes an animal feed additive for preventing diseases caused by a new influenza virus, avian influenza virus and human infectious or acquired common infectious coronavirus containing a Galla Rhois extract or a compound isolated therefrom and the same Provide feed.
  • the animal feed additive composition may be 20 to 90% high concentrate or powder or granule form.
  • composition for animal feed additives of the present invention includes organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate and polyphosphate (polymeric phosphate), polyphenols, and catechins (catechin) ), Alpha-tocopherol, rosemary extract (rosemary extract), vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid and the like may further include any one or more than one.
  • organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate and polyphosphate (polymeric phosphate), polyphenols, and catechins (catechin)
  • Alpha-tocopherol rosemary extract (rosemary extract), vitamin C, green tea extract, licorice extract, chitosan,
  • Animal feed additives containing the composition of the present invention and feed comprising the same as a secondary component various supplements such as amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal enzymes, live microbial preparations, grains, for example Milled or crushed wheat, oats, barley, corn and rice; Vegetable protein feed, such as rape, soybeans and sunflower; Animal protein feeds such as blood meal, meat meal, bone meal and fish meal; Sugars and dairy products, for example, dry powders consisting of various powdered milks and whey powders, and drying additives, and then liquid components and components which become liquids after heating, that is, lipids, for example, animals liquefied by heating, for example.
  • various supplements such as amino acids, inorganic salts, vitamins, antibiotics, antibacterial substances, antioxidants, antifungal enzymes, live microbial preparations, grains, for example Milled or crushed wheat, oats, barley, corn and rice; Vegetable protein feed
  • the animal feed additive composition may be administered to an animal in combination with other feed additives in an edible carrier alone.
  • composition for animal feed additives can be easily administered as a top dressing or directly mixed with the animal feed or separately from the feed, in a separate oral formulation, by injection or transdermal or in combination with other ingredients.
  • a single daily dose or divided daily doses can be used, as is well known in the art.
  • the dosage form of the composition may be prepared in an immediate release or sustained release formulation in combination with these composition-toxic pharmaceutically acceptable edible carriers.
  • edible carriers can be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol.
  • the dosage form of the extract may be tablets, capsules, powders, torokies or sugar-containing tablets or top dressings in microdisperse form.
  • a liquid carrier it may be in the form of soft gelatin capsules or syrups or liquid suspensions, emulsions or solutions.
  • the dosage form may contain auxiliaries such as preservatives, stabilizers, wetting or emulsifying agents, solution promoters and the like.
  • animal feed in which the extract / compound of the present invention is included as an animal feed additive may be any protein-containing organic cereal meal commonly used to meet the dietary needs of animals.
  • protein-containing flours typically consist mainly of corn, soy flour, or corn / bean flour mixtures.
  • the animal feed additive may be used by adding to the animal feed by dipping, spraying or mixing.
  • the invention is applicable to a number of animal diets, including mammals, poultry and fish. More specifically, the diet is commercially important mammals such as pigs, cows, sheep, goats, laboratory rodents (rats, mice, hamsters and gerbils), furry animals (eg mink and fox), and Zoo animals (eg monkeys and tailless monkeys), as well as livestock (eg cats and dogs).
  • Commercially important poultry typically include chickens, turkeys, ducks, geese, pheasants and quails.
  • Commercially raised fish, such as trout may also be included.
  • the composition is incorporated into the animal feed in an amount of about 1 g to 100 g per kg of feed on a dry weight basis.
  • the feed mixture is thoroughly mixed and then a viscous granulated or granular material is obtained depending on the degree of grinding of the components. It is fed as a mash or formed into the desired discrete shape for further processing and packaging. At this time, it is preferable to add water to the animal feed and then go through the usual pelletization, expansion, or extrusion process to prevent separation during storage. Excess water may be removed to dryness.
  • the gall bladder extract of the present invention and the compounds isolated therefrom exhibit excellent antiviral effects against diseases caused by the new influenza virus, avian influenza virus and human infectious or common infectious coronavirus. It can be provided as an additive or feed.
  • FIG. 1 is a diagram showing the anti-influenza virus effect of the five constituent compounds isolated from the gall bladder extract through RT-PCR and plaque assay
  • FIG. 2 is a diagram showing the results of inhibition of RNA synthesis and protein N and M of coronavirus
  • 3 to 5 is a diagram showing the anti-coronavirus effect of OB-34, OB-36, OB-3131 as seen through the Plaque assay,
  • 6 to 8 are antiviral synergistic effects of OB-36 and OB-3131 on coronaviruses MHV-A59 and MHV-JHM, stomatitis VSV,
  • FIG. 9 is a diagram showing the anti-coronavirus action mechanism of OB-34, OB-36, OB-3131.
  • the extract or compound of the present invention may be administered in the following formulations, and the following formulation examples are merely illustrative of the present invention, thereby not limiting the content of the present invention.
  • the above ingredients are mixed and filled in an airtight cloth to prepare a powder.
  • tablets are prepared by tableting according to a conventional method for preparing tablets.
  • the above ingredients are mixed and filled into gelatin capsules to prepare capsules.
  • the amount of the above ingredient is prepared per ampoule (2 ml).
  • each component is added and dissolved in purified water, lemon flavor is added, the above ingredients are mixed, and then, purified water is added to adjust the total amount to 100 ml, and then sterilized by filling into a brown bottle.
  • purified water is added to adjust the total amount to 100 ml, and then sterilized by filling into a brown bottle.
  • Vitamin B6 0.5 mg
  • composition ratio of the vitamin and mineral mixture is a composition suitable for a relatively healthy food in a preferred embodiment
  • the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method.
  • the granules may be prepared and used for preparing a health food composition according to a conventional method.
  • Example 2-1 Take 2.5 g of the 50% ethanol elution fraction obtained in Example 2-1, place it on a reversed phase YMC column and chromatograph using methanol: distilled water (3: 7 ⁇ 5: 5 ⁇ 10: 0, v / v) as the eluent.
  • the chromatography was combined, concentrated and divided into seven fractions (AG).
  • the first fraction, A (0.9 g), was placed on a Sephadex LH-20 column, followed by chloroform and methanol (6). Chromatography was carried out using: 1 ⁇ 3: 1 ⁇ 1: 1, v / v) as the elution solvent, and small fractions (A1 to A3) were separated into three secondary fractions.
  • the first fraction of the second fraction, A1 (30 mg), was subjected to silica gel column chromatography using chloroform: methanol: distilled water (6: 1: 0.1, v / v) using an eluting solvent to give compound 1 (7 mg). It was confirmed that the result of the instrumental analysis was gallic acid (gallic acid) having the following physical properties (Cha et al., Kor. J. Pharmacogn., 31, pp185-189, 2000).
  • A3 (460 mg), the third fraction of the second fraction obtained in Example 2-2, was placed on a Sephadex LH-20 column, and chromatographed using methanol as an elution solvent to give one fraction (A31). Purified. Chromatography using A31 on a reversed-phase YMC column with methanol: distilled water (4: 6, v / v) as the eluent, combining and concentrating those with the same behavior in thin layer chromatography (A311-A314). The third fraction A313 (146 mg) was subjected to Sephadex LH-20 column chromatography using methanol: distilled water (8: 2 ⁇ 9: 1 ⁇ 10: 0, v / v).
  • the gall bladder extract has a killing effect to the concentration of 6.25 mg / ml.
  • influenza virus A / human / korea / KUMC-33 / 2006 / H1N1, distribution institution: Pathogenic Viral Bank [Host, Korea University]
  • the virus was released for a certain time to collect polyethylene glycol (PEG; The virus was precipitated with Polyethylene glycol 8000, Usb corporation) and the viral RNA was extracted.
  • RT-PCR was performed by preparing primers specific to the NS segment, and the virus RNA of the NS segment was detected in the cells treated with only DMSO but untreated group (OB-34, OB-36).
  • OB-311, OB-312, and OB-3131 did not detect the viral RNA of the NS segment, demonstrating its antiviral efficacy against influenza A virus (H1N1) (see FIG. 1). .
  • Influenza virus (A / human / korea / KUMC-33 / 2006 / H1N1, Institution: Pathogenic Virus Bank [Host, Korea University]) after infecting cells with a compound isolated from the blastocyst ethanol extract (OB-34, OB -36, OB-311, OB-312, and OB-3131) were processed together. Cells infected with the virus were further cultured in the incubator for a certain period of time, and then the plaque assay was used to determine whether virus release was reduced by the drug.
  • OB-34, OB -36, OB-311, OB-312, and OB-3131 a compound isolated from the blastocyst ethanol extract
  • influenza A virus H1N1
  • DMSO dimethyl methyl sulfoxide
  • Northern blotting RNA is extracted from virus-infected cells and denatured, followed by electrophoresis on an agarose gel. Thereafter, RNA was detected using a 32 P-labeled random-primed probe specific to the MHV coronavirus strain.
  • Western blotting virus-infected cells are lysed to separate proteins and electrophoresed by SDS-PAGE. Transfer to the membrane to determine the presence or absence of the protein as an antibody to the S, M, N protein of the MHV coronavirus strain.
  • MHV-A59 and MHV-JHM coronavirus strains available from Dr. Shinji Makino, U of Texas Medical Branch, Galveston, TX, USA
  • VSV stomatitis virus strains available from Dr. Shinji Makino, U of Texas Medical Branch, Galveston, TX, USA
  • the compound OB-34, OB-36, OB-311, OB-312, OB-3131
  • Cells infected with the virus were further cultured in the incubator for a certain period of time, and then the plaque assay was used to determine whether virus release was reduced by the drug.
  • OB-34 Three compounds, OB-34, OB-36, and OB-3131, showed up to 1,000 times higher antiviral efficacy against coronavirus compared to the control ribavirin.
  • rhabodoviridae a stranded RNA virus
  • OB-3131 also inhibits the proliferation of VSV (negative strand RNA virus rhabodoviridae) stomatitis virus in addition to coronavirus, confirming its potential as a broad antiviral agent.
  • VSV negative strand RNA virus rhabodoviridae
  • the cells are infected with MHV-A59 and MHV-JHM coronavirus strains and VSV stomatitis virus strains.
  • OB-36 and OB-3131 alone or in half the concentration of the compound separated from the gall bladder extract and gall bladder extract was treated together.
  • Cells infected with the virus were further cultured in the incubator for a certain period of time, and then the plaque assay was used to determine whether virus release was reduced by the drug.
  • Northern blotting RNA is extracted from virus-infected cells and denatured, followed by electrophoresis on an agarose gel. Thereafter, RNA was detected using a 32 P-labeled random-primed probe specific to the MHV coronavirus strain.
  • Western blotting virus-infected cells are lysed to separate proteins and electrophoresed by SDS-PAGE. Transfer to the membrane to determine the presence or absence of the protein as an antibody to the S, M, N protein of the MHV coronavirus strain.
  • OB-34, OB-36, and OB-3131 which are compounds isolated from the gall bladder extract, inhibited the adsorption of the virus to the cells
  • the experimental and untreated controls treated with the compound when the virus was attached to the cells.
  • the degree of adhesion was observed.
  • the degree of adhesion was examined by Northern blotting after isolation of viral RNA in the cell. If the compound inhibits the adsorption of the virus to the cell, the amount of viral RNA will be reduced compared to the control without the compound.
  • OB-34 and OB-36 do not inhibit the RNA synthesis of coronavirus and the synthesis of N protein, a structural protein, and it is highly likely to inhibit the assembly or release of coronavirus, and OB-3131 is a coronavirus. Inhibition of the synthesis of RNA and the synthesis of N protein was likely to inhibit the early stages of coronavirus proliferation (entry, RNA synthesis or processing of nonstructural proteins) (see Fig. 9).
  • the gall bladder extract of the present invention and the compounds isolated therefrom exhibit excellent antiviral effects against diseases caused by the new influenza virus, avian influenza virus and human infectious or common infectious coronavirus. It can be used as an additive or feed.

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Abstract

L'invention concerne un extrait de Galla Rhois présentant des effets inhibiteurs, ou un composé isolé à partir de celui-ci, qui présente des effets antiviraux supérieurs contre des maladies provoquées par de nouveaux virus de la grippe, le virus de la grippe aviaire et le coronavirus qui est infectieux pour l'organisme humain ou habituellement infectieux pour les humains et les animaux, et qui peut donc être utilisé pour des compositions pharmaceutiques, des aliments fonctionnels bons pour la santé, des additifs d'aliments pour animaux ou des aliments pour animaux.
PCT/KR2012/002450 2011-04-19 2012-04-02 Composition contenant un extrait de galla rhois présentant des effets inhibiteur de virus, ou un composé isolé à partir de celui-ci, comme ingrédient actif et ses utilisations WO2012144754A2 (fr)

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CN111658631A (zh) * 2020-06-11 2020-09-15 广东盛普生命科技有限公司 没食子酸及其衍生物和结构类似物在制备抗冠状病毒药物方面的应用
WO2021155864A1 (fr) * 2020-02-08 2021-08-12 Syneurx International (Taiwan) Corp. Composés et leurs utilisations pharmaceutiques
WO2021207399A1 (fr) * 2020-04-07 2021-10-14 Sytheon Limited Tanins hydrolysables destinés à être utilisés dans l'atténuation du syndrome de détresse respiratoire aiguë
TWI803858B (zh) * 2020-04-23 2023-06-01 心悅生醫股份有限公司 化合物及其醫藥用途

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WO2023038480A1 (fr) * 2021-09-08 2023-03-16 에이피알지 주식회사 Composition pour la prévention, le traitement ou le soulagement d'une infection par le virus de la grippe, comprenant un mélange d'extrait d'agrimonia pilosa et d'extrait de galla rhois en tant que principes actifs
KR102420201B1 (ko) 2022-01-17 2022-07-13 주식회사 유이케미칼 인지질 외피 보유 바이러스 사멸기능을 가지는 친환경 방역 조성물
KR102679048B1 (ko) * 2022-04-18 2024-07-01 (주)유제이 유해 조류 기피제 조성물
KR20240045546A (ko) 2022-09-30 2024-04-08 (주)예스킨 항 인플루엔자 바이러스제

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WO2021155864A1 (fr) * 2020-02-08 2021-08-12 Syneurx International (Taiwan) Corp. Composés et leurs utilisations pharmaceutiques
US11154531B2 (en) 2020-02-08 2021-10-26 Syneurx International (Taiwan) Corp. Compounds and pharmaceutical uses thereof
US11382924B2 (en) 2020-02-08 2022-07-12 Syneurx International (Taiwan) Corp. Compounds and pharmaceutical uses thereof
US11779561B2 (en) 2020-02-08 2023-10-10 Syneurx International (Taiwan) Corp. Compounds and pharmaceutical uses thereof
EP4103539A4 (fr) * 2020-02-08 2024-05-29 Syneurx International (Taiwan) Corp. Composés et leurs utilisations pharmaceutiques
WO2021207399A1 (fr) * 2020-04-07 2021-10-14 Sytheon Limited Tanins hydrolysables destinés à être utilisés dans l'atténuation du syndrome de détresse respiratoire aiguë
WO2021207325A1 (fr) * 2020-04-07 2021-10-14 Sytheon Limited Extraits naturels et leurs composants destinés à être utilisés dans l'atténuation du syndrome de détresse respiratoire aiguë
TWI803858B (zh) * 2020-04-23 2023-06-01 心悅生醫股份有限公司 化合物及其醫藥用途
US11793823B2 (en) 2020-04-23 2023-10-24 Syneurx International (Taiwan) Corp. Compounds and pharmaceutical uses thereof
CN111574571A (zh) * 2020-06-11 2020-08-25 石河子大学 一种五倍子有效部位的分离方法及其用途
CN111658631A (zh) * 2020-06-11 2020-09-15 广东盛普生命科技有限公司 没食子酸及其衍生物和结构类似物在制备抗冠状病毒药物方面的应用
CN112336709A (zh) * 2020-06-11 2021-02-09 广东盛普生命科技有限公司 没食子酸及其衍生物和结构类似物在制备抗冠状病毒药物方面的应用

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