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WO2007099915A1 - Agent antiviral - Google Patents

Agent antiviral Download PDF

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Publication number
WO2007099915A1
WO2007099915A1 PCT/JP2007/053538 JP2007053538W WO2007099915A1 WO 2007099915 A1 WO2007099915 A1 WO 2007099915A1 JP 2007053538 W JP2007053538 W JP 2007053538W WO 2007099915 A1 WO2007099915 A1 WO 2007099915A1
Authority
WO
WIPO (PCT)
Prior art keywords
hop
antiviral agent
cold water
water extract
influenza virus
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/JP2007/053538
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English (en)
Japanese (ja)
Inventor
Syuichi Segawa
Hisako Yasui
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.)
Sapporo Breweries Ltd
Shinshu University NUC
Original Assignee
Sapporo Breweries Ltd
Shinshu University NUC
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Sapporo Breweries Ltd, Shinshu University NUC filed Critical Sapporo Breweries Ltd
Priority to JP2008502775A priority Critical patent/JP5171614B2/ja
Publication of WO2007099915A1 publication Critical patent/WO2007099915A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • 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)
    • A61K36/348—Cannabaceae
    • A61K36/3486—Humulus
    • 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)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • 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
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Preparation or treatment thereof
    • A23L2/52—Adding ingredients
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • 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
    • 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

Definitions

  • the present invention relates to an antiviral agent.
  • EGCG green tea component epicarocatechin gallate
  • HTFDG tea component theaflavin galley HTFDG
  • Hops include antioxidant bituminous glycosides such as humulone and lubron (Patent Literatures 1 to 3), and flavonoid glycosides such as rutin and isoquercitrin. It has been reported that it can be used as an antioxidant for foods and beverages and cosmetics (Patent Document 4).
  • Patent Document 1 Japanese Patent Laid-Open No. 04-202138
  • Patent Document 2 Japanese Patent Laid-Open No. 06-025081
  • Patent Document 3 Japanese Patent Laid-Open No. 06-312924
  • Patent Document 4 Japanese Patent Application Laid-Open No. 09-2917
  • Non-Patent Document 1 Nakayama et al., 1996, Journal of Infectious Diseases, 70 ⁇ , No. 11, 1190-1192
  • Non-Patent Document 2 Nakayama et al., 1996, Journal of Infectious Diseases, 68 ⁇ , No. 7, p824-828 Disclosure of the invention
  • hop pellets used for brewing effervescent alcoholic beverages such as beer (referred to dried hop blossoms, sieved after pulverization, and solidified after passing through a sieve)
  • hop extract extract extracted from hop blossoms, extracted from organic solvent or supercritical fluid
  • brewing sparkling alcoholic beverages such as beer Tissue or hop residues
  • secondary use of unused hops or hop residues that are discarded is not profitable in terms of cost, and there is a demand for higher usage and usage!
  • an object of the present invention is to provide a highly safe antiviral agent derived from a plant. Another object of the present invention is to provide foods and drinks, feeds and feed additives containing the above-mentioned antiviral agents, and pharmaceuticals containing the above antiviral agents as active ingredients. Means for solving the problem
  • the present invention provides an antiviral agent characterized by comprising a cold water extract of hop tissue as an active ingredient.
  • the extract of hop tissue has a cold water extract strength, particularly effectively suppressing chicken hemagglutination caused by influenza virus and suppressing viral infection of animal cells.
  • Hops are mainly used for brewing beer and have long been used for various purposes other than brewing. Therefore, anti-viral agents derived from hops are considered to have excellent safety to the human body.
  • the antiviral agent of the present invention is obtained by extracting hop tissue with cold water, it is not affected by heat and can stably maintain the activity of the antiviral agent.
  • the hop structure is a pulverized product of dried hop camellia, and is also a pulverized product of dried hop camellia, which is at least part of a pulverized product having a size equal to or less than the size of lupulin. It is preferable that is removed.
  • the hops used in brewing effervescent alcoholic beverages such as beer are the power to dry coconuts excluding stems and leaves, sieve them after pulverization, solidify those that have passed through the sieves, and use them as hop pellets The crushed material that does not pass through the sieve is discarded.
  • This discarded pulverized product is obtained by removing at least a part of the pulverized product having a size smaller than that of Lubrin, and most of it is hop cake. Contributes to the reduction of industrial waste and makes effective use of hop dredging.
  • the hop tissue may be a hop residue obtained by removing at least a part of a substance extracted from an organic solvent or a supercritical fluid from a dried hop spikelet.
  • hop spikelets used in beer brewing are used for hop extract extraction, but the hop residue remaining after hop extract extraction is discarded.
  • This discarded hop residue is obtained by removing at least a part of the substance extracted by organic solvent extraction or supercritical fluid, so if this is used as a raw material for extraction of antiviral agents, industrial waste will be reduced. Can contribute.
  • the dried hop spikelet pulverized product is preferably a pulverized product of dried hop spikelet frozen product.
  • the efficiency of the pulverization increases and the effect of heat generated during the pulverization is reduced, so that the activity of the antiviral agent can be stably maintained.
  • the pulverized product having a size smaller than the size of rubulin easily passes through the sieve, the purity of the pulverized product exceeding the size of rubulin is increased, and the purity of the component that affects the antiviral activity is also increased. it can.
  • the antiviral agent of the present invention includes astragalin, astragalin malonyl darcoside, isocercitrin, isoquercitrin malo rudarcoside, quercetin malo rudarcoside, kaempferol rutinoside, kenferrol malo rudarcoside, rutin And at least one, preferably at least two, more preferably all flavonoid glycosides selected from the group consisting of fluorosilenone glycosides.
  • the antiviral agent of the present invention preferably contains 0.001% to 5% by weight of each of these flavonoid glycosides.
  • the fluorosilphenone glycoside includes fluoroisobutyral phenone dalcoside, fluoro-2-methylbutyrophenone dalcoside, and fluoroisovale phenone dalcoside.
  • Influenza virus growth begins with the binding of hemagglutinin (hemadaltun protein) to the surface receptors of host cells.
  • hemagglutinin hemadaltun protein
  • Glycoproteins or glycolipids with N-acetylneuraminic acid serve as receptors.
  • the antiviral agent of the present invention contains at least one of the above flavonoid glycosides, it can be expected to be adsorbed on the hemagglutinin protein of influenza virus, and for this reason, it can be adsorbed to the cell surface receptor of influenza virus. It is thought that the binding is inhibited and the entry of influenza virus into the cell is prevented.
  • the antiviral agent is preferably an anti-influenza virus agent.
  • the antiviral agent Since the antiviral agent has an activity of suppressing the influenza hemagglutination reaction of influenza virus and inhibiting the infection of animal cells, it prevents infection of influenza virus in humans and animals, especially poultry, particularly chickens. It is suitable for preventing.
  • the antiviral agent of the present invention described above is excellent in antiviral activity and safety, it can be used as an active ingredient by being contained in foods, drinks, feeds, feed additives or pharmaceuticals.
  • the invention's effect is excellent in antiviral activity and safety, it can be used as an active ingredient by being contained in foods, drinks, feeds, feed additives or pharmaceuticals. The invention's effect
  • an antiviral agent which has an activity of suppressing viral chicken platelet aggregation reaction and can prevent infection of humans and animals with pathogenic viruses. Since the antiviral agent of the present invention is derived from a natural plant, it is excellent in safety, and each person can prevent infection with pathogenic virus in the home and lead to early healing after infection.
  • the antiviral agent of the present invention is produced as a by-product in the brewing of effervescent alcoholic beverages such as beer, and can use hops discarded as raw materials, contributing to the reduction of industrial waste, Can be used effectively by increasing the added value.
  • FIG. 1 is a simplified diagram showing the procedure of a chicken erythrocyte aggregation titer measurement test.
  • FIG. 2 is a graph showing the inhibitory action of a hop pellet cold water extract and a bent hop cold water extract on the hemagglutination action of influenza virus.
  • FIG. 3 is a diagram schematically showing the procedure of an infection inhibition test for animal cells of influenza virus.
  • FIG. 4 is a graph showing the inhibitory action of hop pellet cold water extract and Svent hop cold water extract on infection of influenza virus to MDCK cells.
  • FIG. 6 is a graph showing the infection rate of influenza virus in mice of each group.
  • FIG. 7 shows the survival rate of mice in each group.
  • the hop used in the present invention may be any hop varieties, but beer brewing hop varieties such as Czech Zapp, German Haratau tradition and domestic Furano No. 18 are preferred. In order to obtain a hop extract with a high antiviral activity, it is more preferable than the Czech zazat.
  • the hop organization in the present invention means any hop organization or a part thereof.
  • the hop tissue used for cold water extraction is more preferably a hop cocoon that prefers a cocoon that uses any of leaves, stems, and spikelets.
  • the hop bud is a cocoon leaf constituting the cocoon flower, and can be obtained by removing at least a part of the rubulin portion (yellow granule) from the cocoon flower.
  • the hop tissue used for cold water extraction of the present invention may be a hop koji that is discarded without being pulverized to a specified size when processing hop pellets used for brewing foaming alcoholic beverages such as beer. It may be a hop residue remaining after the hop blossoms described later are extracted with a supercritical fluid or an organic solvent.
  • the antiviral agent of the present invention contains a cold water extract of hop tissue as an active ingredient, and this active ingredient is obtained by a production method comprising a step of extracting a hop tissue with cold water.
  • the hop tissue is extracted with cold water
  • “cold water” means water at room temperature or lower, and usually refers to water at temperatures above 0 ° C and below 30 ° C.
  • the temperature of chilled water is preferably 5 ⁇ 3 ° C (more preferably 5 ⁇ 2 ° C), more preferably over 0 ° C and below 10 ° C.
  • extraction efficiency can be increased by adding a small amount of alcohol, preferably ethanol, to 10% by weight or less to water.
  • Hop tissue strength As a method of extracting an antiviral agent, natural products are extracted from plants with water. For example, there may be mentioned a method in which a hop structure and a certain amount of cold water are put in a container, allowed to stand for a predetermined time with appropriate stirring, and the extract is filtered to remove the residue. In order to completely remove impurities and impurities contained in the mixture, the supernatant obtained by further centrifuging the filtered extract (hereinafter, centrifuged supernatant) can be used as an antiviral agent. In addition, the obtained antiviral agent can be concentrated and dried for use.
  • the antiviral agent which is a cold water extract of hop tissue
  • a synthetic adsorbent examples include Amberlit e XAD-4, 7 and 16 (organo), activated carbon, and polybulupolypyrrolidone (PVPP; polyphenol adsorbent).
  • Amberlite XAD-4 is preferably used.
  • the water extract of hop tissue is passed through a column packed with a synthetic adsorbent, and the adsorbed component is eluted with, for example, a mixed solvent of water and methanol, and the eluted fraction is used as an antiviral agent. Can be used.
  • the antiviral agent of the present invention comprises a cold water extract of dried hop koji crushed material as an active ingredient. It is preferable to use a cold water extract from which at least a part of the pulverized product is removed as an active ingredient.
  • the pulverized product of dried hop spikelets used for cold water extraction includes, for example, a drying step of drying hop spikelets to obtain dried hop spikelets, and a pulverization step of pulverizing dried hop spikelets to obtain a pulverized product. And a sorting step for removing pulverized material having a size equal to or smaller than the size of lupulin.
  • the hop spikelets are dried at a temperature of 100 ° C or lower and the water content can be removed to such an extent that the hop spikelets can be stored. It is preferred to dry to ⁇ 9%.
  • the hop spikelets can be efficiently pulverized into a fine powder.
  • a pulverizer such as a pin mill, a hammer mill, or a ball mill may be used.
  • the sorting step the pulverized product is sieved, and for example, a pulverized product having a major axis of 0.1 mm or more can be selected as “larger than Lubrin”.
  • the major axis not to pass through the sieve to a major axis of 0.3 mm or more, more preferably a major axis of 0.5 mm or more.
  • a major axis of 0.5 mm or more it is more preferable to set the major axis not to pass through the sieve to a major axis of 0.3 mm or more, more preferably a major axis of 0.5 mm or more.
  • the cold water extract of the dried hop camellia powder obtained by removing at least a part of the pulverized product having a size smaller than the size of Lubrin is extracted with the above-mentioned cold water. Extract it using the method described in the step.
  • the dried hop spikelet pulverized product used for preparing the antiviral agent of the present invention is preferably a dried hop spikelet frozen product pulverized product.
  • the method for freezing dried hop spikelets is not particularly limited, but is preferably 10 ° C or lower, more preferably 35 ° C or lower.
  • the antiviral agent of the present invention is a hop residue cold water extract obtained by removing at least part of a substance extracted from an organic solvent or a supercritical fluid from dried hop spikelets. It can be.
  • the organic solvent used for the organic solvent extraction include alcohol and hexane, and ethanol having lower alcohol having 1 to 4 carbon atoms is more preferable.
  • the supercritical fluid used for the supercritical fluid extraction include carbon dioxide, water, methane, ethane, ethylene, propane, pentane, methanol, and ethanol, with carbon dioxide and carbon being preferred.
  • the antiviral agent of the present invention has an effect of preventing infection of various pathogenic viruses, and is suitable for preventing infection of airborne viruses.
  • airborne viruses include rhinovirus, coronavirus, adenovirus, enterovirus, influenza virus, parainfluenza virus, SARS coronavirus, RS virus, echovirus, simple herpes virus, EB virus, Examples include measles virus and cytomegalovirus, which are more suitable for preventing influenza virus infection.
  • the antiviral agent of the present invention can be added to foods and drinks, feeds or feed additives for the purpose of preventing infection with pathogenic viruses and alleviating symptoms, or preventing infection with pathogenic viruses or It can be used as an active ingredient of a pharmaceutical agent as a therapeutic agent.
  • food and drink refers to food and drink consumed by humans
  • feed refers to food and drink consumed by animals other than humans.
  • feed additive refers to what is added to such “feed”.
  • the above-mentioned food and drink, feed, feed additive and medicine have a hop tissue cold water extract.
  • a cold-hop extract of hop tissue or a freeze-dried product of this cold-water extract can be used as it is as the food, feed, feed additive or pharmaceutical product.
  • the food / beverage product, feed, and feed additive of the present invention contain the antiviral agent described above, and may contain an additive that is usually used in the field.
  • the additives include apple fiber, soybean fiber, meat extract, black vinegar extract, gelatin, corn starch, honey, animal and vegetable oils and fats, monosaccharides such as glucose, disaccharides such as sucrose, fructose and mannitol, dextrose and Examples include polysaccharides such as starch, sugar alcohols such as erythritol, xylitol and sorbitol, and vitamins such as vitamin C. These additives may be used alone or in combination.
  • the pharmaceutical agent of the present invention contains the above-described antiviral agent as an active ingredient, and may further contain a pharmaceutically acceptable additive.
  • the additives include monosaccharides such as glucose, disaccharides such as sucrose, fructose and mannitol, polysaccharides such as dextrose and starch, sugar alcohols such as erythritol, xylitol and sorbitol, vitamin C and the like.
  • Vitamins may be a single species or a plurality of species.
  • rub pellets rich in rubulin produced from dried hop spikelets
  • Svent hop force which is a fraction that has been partially removed, prepared cold water extracts (hereinafter referred to as “hop pellet cold water extract” and “svent hop cold water extract”, respectively).
  • the hops are dried at 50 ° C until the water content becomes 8%, pulverized with a dedicated pulverizer, and the hop structure that has passed through a sieve with a mesh opening of 0.3 mm is made into hop pellets, and this sieve is passed through. None, 0.3 mm or more yarn and weave was used as a vent hop. That is, the hop pellet is a fraction containing fine powder of hop tissue and rubulin, and the hop cocoon is a pulverized product of 0.3 mm or more of the cocoon leaf part constituting the coconut flower, and the pudding is removed from the pudding. Minutes.
  • the hop pellets or the bent hops thus obtained were suspended in cold water so as to be 5%, and left in a low-temperature room at 4 ° C for 1 hour to extract cold water. Thereafter, each cold water extract was filtered (ADVANTEC No. 5A) to remove the residue and freeze-dried.
  • Hop pellet cold water extract and Svent hop cold water extract are quantified for polyphenol concentration according to the foreign thiocult method shown below, and based on this concentration, chicken hemagglutination assay described later and infection of animal cells with influenza virus An inhibition test was conducted.
  • each cold water extract was dissolved in a phosphate buffer solution (hereinafter, 0.1% BSA-PBS) containing 0.1% ushi serum albumin so as to have a concentration of 0.05 mgZmL.
  • the phenol reagent was removed and allowed to react for 3 minutes.
  • an equivalent amount of 10% aqueous sodium carbonate solution was added to the added 1N phenol, and the mixture was further reacted at room temperature for 1 hour, and the absorbance at 700 nm was measured.
  • a calibration curve was prepared using a standard solution of gallic acid whose concentration was previously divided, and the concentration of polyphenol in each cold water extract was calculated as the gallic acid equivalent from this calibration curve.
  • Example 1 Inhibition of erythrocyte aggregation by cold water extract of hop tissue:
  • Influenza virus has hemagglutinin (HA), and has the effect of agglutinating erythrocytes in birds. Therefore, the presence or absence of influenza virus in the sample can be determined by examining the presence or absence of chicken erythrocyte agglutination.
  • the infection and propagation of influenza virus in humans and animals begins with the binding of viral surface HA to receptors on the host cell surface. Host details Influenza virus adsorbed on the cell surface is taken up into the cell by endocytosis, grows in the cell using the translation system of the host cell, and repeatedly infects other cells. For this reason, it is thought that if the binding of influenza virus to animal cells via HA is suppressed, infection of humans and animals with influenza virus can be prevented. So, hop yarn and
  • the following chicken erythrocyte agglutination titration measurement test was conducted to determine whether or not the woven cold water extract had an inhibitory effect on chicken erythrocyte aggregation.
  • Hop pellet cold water extract and Svent hop cold water extract are each thread and weave cold water extraction as described above, 0.1% BS so that the polyphenol concentration becomes a predetermined concentration after lyophilization.
  • the erythrocyte suspension was prepared by suspending chicken stock blood in 0.1% BSA'PBS, washing the erythrocytes by repeating the centrifugation at 3000rpm for 5 minutes three times, and then adding the precipitated erythrocyte fraction to the sedimented erythrocyte fraction. for
  • a new 0.1% BSA'PBS was prepared so as to be 0.5% (vZv).
  • the AZPuerto RicoZ8Z34 (PR8: HlNl) strain was used as the influenza virus strain.
  • Influenza virus solution is diluted with 0.1% BSA'PBS so that the TCID value is 0 5 8
  • TCID value causes virus infection in 50% of cultured cells
  • Fig. 1 is a simplified diagram showing the procedure of the chicken hemagglutination titer measurement test.
  • First add 50 ⁇ L of influenza virus solution and 50 ⁇ L of hop pellet cold water extract, subvent hop cold water extract, or 0.1% BSA'PBS to each well of a 96-well microplate, The reaction was allowed to proceed for 1 hour at room temperature. Thereafter, 2-fold serial dilution series (2 1-fold dilution - 2 15-fold dilution) made for each group by micropipette, these 50 L of 0. 5% (VZV) erythrocyte suspension at room temperature was added respectively The sample was allowed to stand for 1 hour, and the presence or absence of an agglutination image was observed.
  • VZV 0. 5%
  • the chicken hemagglutination reaction was determined to be positive when an hemagglutination image was observed, and the maximum dilution ratio at which the hemagglutination reaction was positive was determined as the hemagglutination titer (HA titer). Red
  • the presence or absence of hemagglutination-inhibiting activity is determined by comparing the HA titer (control group) of the dilution series obtained by reacting the erythrocyte suspension with a mixture of influenza virus solution and 0.1% BSA'PBS and the HA titer of each test extract. It investigated by comparing.
  • the values in the graph are the mean standard error of each experiment performed three times, and the Mann-Whitney test was used to test the significant difference between the HA value of the control group and the HA value of the test group.
  • FIG. 2 is a graph showing the hemagglutination-inhibiting action of the hop pellet cold water extract and the bent hop cold water extract on the HA titer of influenza virus.
  • the vertical axis represents the HA value, and the concentration of each test extract is represented by the polyphenol content.
  • the hop pellet cold water extract and the scavenged hop cold water extract significantly suppressed the HA titer of influenza virus at a concentration of 0.6 mg / mL. From this result, it was found that the cold tissue extract of hop tissue contains a substance that suppresses the hemagglutination reaction of influenza virus, suggesting that it has antiviral activity.
  • MDCK cells Melat-Darby canine kidney cells
  • MDCK cells are suitable for various virus infection experiments and are generally used for evaluating the infection inhibitory action of influenza virus.
  • the cells were cultured in MEM medium containing serum (FBS) and maintained by subculture twice a week.
  • FBS serum
  • a cell suspension of MDCK cells prepared in 3 ⁇ 10 5 cells ZmL was added to a 96-well plate at 200 ⁇ L, and cultured for 3 or 4 days.
  • Fig. 3 is a simplified diagram showing the procedure for an infection inhibition test for animal cells of influenza virus.
  • a 25 L influenza virus dilution (the above influenza The virus solution was diluted 5 ⁇ 10 4 times) and each concentration of hop pellet cold water extract or spent hop cold water extract was mixed in a 96-well plate and allowed to react at room temperature for 1 hour (hereinafter referred to as “this”).
  • the reaction solution is called a virus reaction solution;).
  • the MDCK cell force MEM medium previously cultured for 3 days or 4 days in a 96-well plate is removed by aspiration, and 25 ⁇ L of the virus reaction solution is added to the medium, and it is added in a 37 ° C CO incubator. The reaction was further continued for 1 hour.
  • maintenance medium (MEM medium containing 0.2% albumin and 5 gZmL acetyltilpsin) is added to each tool and cultured for 3 days.
  • the culture supernatant has the ability to agglutinate chicken erythrocytes. I investigated whether or not.
  • FIG. 4 is a graph showing the inhibitory action of the hop pellet cold water extract and the bent hop cold water extract on the infection of MDCK cells with influenza virus.
  • the vertical axis represents chicken hemagglutination inhibition rate (%), and the concentration of each test extract is represented by the polyphenol content.
  • the hop pellet cold water extract and the bent hop cold water extract inhibit infection and proliferation of MDCK cells by reacting with influenza virus for 1 hour at room temperature.
  • the benthic hop cold water extract has a concentration of 50 / z gZmL Inhibits more than 80% of infections and is stronger than hop pellet cold water extract at the same concentration.
  • the TCID of the subvent hop cold water extract was calculated to be 28.6 ⁇ gZmL.
  • Fig. 5 shows the results of LC MS analysis of flavonoids contained in the bent-hop chilled water extract.
  • astragalin, isoquercitrin, quercetin malonyl darcoside, kaempferol rutinoside, kenferrol malo-aldarcoside and rutin and the fluoroacylphenone glycoside, fluoroisobutyrophenone darcoside, fluoro-2-methylbutyrate. It was found to contain oral phenone dalcoside and fluoroisovalerophenone darcoside.
  • HPLC analysis conditions are as follows.
  • Table 2 shows the results of comparing the content ratios of quercetin and kaempferol before and after hydrolysis of the subvent hop cold water extract (%; weight ratio of each flavonoid aglycone to the dry weight of the hop cold water extract). Is.
  • the flavonoid aglycone containing quercetin and kaempferol was not detected before hydrolysis. After hydrolysis, it contained 1.02% quercetin and 1.21% kaempferol. There was found. Since the flavonoid aglycone was not detected in the Sventhop cold water extract before hydrolysis, the flavonoids contained in the Sventhop cold water extract exist as glycosides, contributing to antiviral activity! / It was suggested to speak.
  • the hop pellet cold water extract administration group contains a 0.1% BSA'PBS solution (hop pellet cold water extract administration solution) containing a hop pellet cold water extract and influenza virus, 0.1% BSA'PBS solution (svent hop cold water extract administration solution) containing Sventhop cold water extract and influenza virus, and the control group containing only influenza virus 0.1% BSA'PBS solution ( A control influenza virus administration solution) was administered to the nostrils of each mouse.
  • Influenza virus solution prepared by diluting 10 times with 1% BSAZPBS was used for the experiment.
  • the hop pellet cold water extract and the scavent hop cold water extract were prepared as described above, and dissolved in 0.1% BSAZPBS so that the polyphenol concentration was 50 mgZmL.
  • the hop pellet cold water extract administration solution and the subvent hop cold water extract administration solution used for infection of mice are 1. 98 mL of the hop pellet cold water extract and the spent hop cold water extract, respectively, and the above influenza virus solution. Were added at 0.02 mL each and reacted at 37 ° C for 3 minutes. A control solution for influenza virus! The above influenza virus solution was added to 0.1% BSAZPBS containing no hop tissue cold water extract and reacted at 37 ° C for 3 minutes.
  • mice were performed according to the following procedure. First, Nembutal was administered intraperitoneally to anesthetize the mice, and a hop pellet cold water extract administration solution and a spout were placed in both nostrils of each group of mice. 10 ⁇ L each (20 ⁇ LZ mouse) of the administration solution of the top hop cold water extract or the control influenza virus was administered. Every day from the start of administration, the changes in body weight of mice were recorded and observed for irregularities in hair and decreased motility, and the presence or absence of influenza virus infection and the number of survivors were examined. The presence or absence of influenza virus infection was determined to be established when two or more items were met, based on changes in mouse body weight, irregularities in hair, and decreased mobility.
  • FIG. 6 shows the infection rate of influenza virus in mice in each group
  • FIG. 7 shows the survival rate of mice in each group. The lower the infection rate and the higher the survival rate, the more the influenza virus infection was suppressed.
  • the hop pellet cold water extract administration group no infection with influenza virus was observed over the 2 weeks of the study period, and the survival rate was 100%.
  • the survival rate of the hop pellet cold water extract group was significantly higher than that of the control group, and a statistically significant difference ( ⁇ ⁇ 0.01) was observed between the two groups.
  • mice were observed on the third day of administration, and all mice were infected with influenza virus on the eighth day of administration. It was confirmed that the death of all mice did not die as in the control group, and 3 mice were alive even after 2 weeks.
  • the group treated with the cold pellet extract showed a higher survival rate compared to the control group, and a statistically significant difference was observed between the two groups ( ⁇ 0. 05).
  • the hop pellet cold water extract and the bent hop cold water extract effectively inhibit influenza virus infection.
  • Industrial applicability ADVANTAGE OF THE INVENTION According to this invention, it has the activity which suppresses the chicken platelet aggregation reaction of a virus, and the antiviral agent which can prevent the infection to the human and animal of pathogenic virus is provided. Since the antiviral agent of the present invention is derived from a natural plant, it is excellent in safety, and since it is produced as a by-product when brewing foaming alcoholic beverages such as beer and discarded, hops are used as raw materials. It contributes to the reduction of industrial waste and can be used effectively with increased added value of hops.

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Abstract

La présente invention concerne un agent antiviral doté d'une grande innocuité, provenant d'une plante, d'un aliment et d'une boisson contenant cet agent antiviral, ainsi qu'un médicament contenant ledit agent en tant que principe actif. La présente invention a trait à un agent antiviral caractérisé en ce qu'il contient, en tant que principe actif, un extrait à l'eau froide d'un tissu de houblon; l'invention a également trait à des aliments, des boissons, des aliments pour animaux, des additifs pour aliments pour animaux ou un médicament caractérisé en ce qu'ils contiennent ledit agent antiviral.
PCT/JP2007/053538 2006-03-01 2007-02-26 Agent antiviral Ceased WO2007099915A1 (fr)

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009093533A1 (fr) * 2008-01-24 2009-07-30 Sapporo Breweries Limited INHIBITEUR DE LA PRODUCTION D'ANTICORPS IgE
JP2010094064A (ja) * 2008-10-15 2010-04-30 Sapporo Breweries Ltd 生地用添加剤及びこれを使用した方法
JP2010173942A (ja) * 2009-01-27 2010-08-12 Sapporo Breweries Ltd 脂肪細胞分化抑制剤
JP2011083266A (ja) * 2009-10-19 2011-04-28 Taiyu Shinko Kosha:Kk ホップ組成物及びその製造方法
CN103145783A (zh) * 2013-03-26 2013-06-12 靖宇县金翔农林生物科技有限公司 用返魂草提取异槲皮苷、返魂草甙、返魂草多糖的方法
WO2014103011A1 (fr) * 2012-12-28 2014-07-03 サントリーホールディングス株式会社 Boisson au goût de bière non alcoolisée ayant un goût acidulé
DE102015115876A1 (de) * 2015-09-21 2017-03-23 Eberhard Karls Universität Tübingen Medizinische Fakultät Substanz zur Prophylaxe und Behandlung von Infektionen durch Influenzaviren
WO2023276811A1 (fr) 2021-07-02 2023-01-05 東洋精糖株式会社 Agent inhibiteur de l'invasion cellulaire par un virus

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03101623A (ja) * 1989-09-14 1991-04-26 Mitsui Norin Kk インフルエンザウィルス感染予防剤
JPH0698738A (ja) * 1992-01-20 1994-04-12 Asama Kasei Kk 食品用保存剤
JPH092917A (ja) * 1995-06-19 1997-01-07 Asahi Breweries Ltd ホップより得られるポリフェノール製剤とその製造法
JP2002505267A (ja) * 1998-03-05 2002-02-19 メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフトング 抗ウイルス性効果をもつ製剤

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03101623A (ja) * 1989-09-14 1991-04-26 Mitsui Norin Kk インフルエンザウィルス感染予防剤
JPH0698738A (ja) * 1992-01-20 1994-04-12 Asama Kasei Kk 食品用保存剤
JPH092917A (ja) * 1995-06-19 1997-01-07 Asahi Breweries Ltd ホップより得られるポリフェノール製剤とその製造法
JP2002505267A (ja) * 1998-03-05 2002-02-19 メルク パテント ゲゼルシャフト ミット ベシュレンクテル ハフトング 抗ウイルス性効果をもつ製剤

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
BARTOSZCZE M.: "ANTI VIRAL PROPERTIES OF PLANT EXTRACTS ACTIVITY OF HOP-D CONES STROBULI LUPULI-D", BULLETIN OF THE VETERINARY INSTITUTE IN PULAWY, vol. 12, no. 1-4, 1968, pages 69 - 75, XP003017064 *
BUCKWOLD V.E. ET AL.: "Antiviral activity of hop constituents against a series of DNA and RNA viruses", ANTIVIRAL RESEARCH, vol. 61, no. 1, 2004, pages 57 - 62, XP002398448 *
COLLECTION OF CZECHOSLOVAK CHEMICAL COMMUNICATIONS, vol. 29, no. 5, 1964, pages 1259 - 1265 *
DATABASE CA [online] HUBACEK J. ET AL.: "Paper chromatography of flavanol glycosides of hops", XP003017067, accession no. STN Database accession no. (1964:92487) *
KURUMATANI M. ET AL.: "Analysis of Polyphenols from Hop Bract Region Using CCC", J. LIQ. CHROMATOGR. RELAT TECHNOL., vol. 28, no. 12/13, 2005, pages 1971 - 1983, XP003017066 *
MITROCOTSA D. ET AL.: "Evaluation of the antiviral activity of kaempferol and its glycosides against human cytomegalovirus", PLANTA MEDICA, vol. 66, no. 4, 2000, pages 377 - 379, XP003017069 *
WANG Q. ET AL.: "Xanthohumol, a novel anti-HIV-1 agent purified from hops Humulus lupulus", ANTIVIRAL RESEARCH, vol. 64, no. 3, 2004, pages 189 - 194, XP004646261 *
WEI F. ET AL.: "Antiviral Flavonoids from the Seeds of Aesculus chinensis", JOURNAL OF NATURAL PRODUCTS, vol. 67, no. 4, 2004, pages 650 - 653, XP003017068 *
YAMAMOTO H.: "Shokuhin Seizo Kotei kara Shojiru Haikibutsu no Yuka Busshitsu eno Tenkan Saisei Gijutsu Project Hop-ho Bubun Kara no Polyphenol-rui no Chushutsu Gijutsu no Kaihatsu", KANSAI UNIVERSITY SANGAKU RENKEI KENKYU CENTER KENKYU SEIKA HOKOKUSHO HEISEI 16 NENDO, 2004, pages 58 - 61, XP003017065 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009093533A1 (fr) * 2008-01-24 2009-07-30 Sapporo Breweries Limited INHIBITEUR DE LA PRODUCTION D'ANTICORPS IgE
JP2010094064A (ja) * 2008-10-15 2010-04-30 Sapporo Breweries Ltd 生地用添加剤及びこれを使用した方法
JP2010173942A (ja) * 2009-01-27 2010-08-12 Sapporo Breweries Ltd 脂肪細胞分化抑制剤
JP2011083266A (ja) * 2009-10-19 2011-04-28 Taiyu Shinko Kosha:Kk ホップ組成物及びその製造方法
WO2014103011A1 (fr) * 2012-12-28 2014-07-03 サントリーホールディングス株式会社 Boisson au goût de bière non alcoolisée ayant un goût acidulé
US10993460B2 (en) 2012-12-28 2021-05-04 Suntory Holdings Limited Non-alcohol, beer-taste beverage having Shimari in taste
CN103145783A (zh) * 2013-03-26 2013-06-12 靖宇县金翔农林生物科技有限公司 用返魂草提取异槲皮苷、返魂草甙、返魂草多糖的方法
DE102015115876A1 (de) * 2015-09-21 2017-03-23 Eberhard Karls Universität Tübingen Medizinische Fakultät Substanz zur Prophylaxe und Behandlung von Infektionen durch Influenzaviren
WO2023276811A1 (fr) 2021-07-02 2023-01-05 東洋精糖株式会社 Agent inhibiteur de l'invasion cellulaire par un virus
KR20240031963A (ko) 2021-07-02 2024-03-08 토요 슈가 리파이닝 컴퍼니 리미티드 바이러스의 세포 침입 저해제

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