JP2010120951A - Anti-influenza virus agent, article, beverage and food for suppressing infection of influenza by adsorbing, impregnating and adding the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an article, beverage and food for suppressing infection of influenza by adsorbing, impregnating and adding an anti-influenza virus agent expressing high suppression effect against infection of influenza virus and having no adverse effect by using a highly safe plant extract allowing daily use without anxiety. <P>SOLUTION: The anti-influenza virus agent comprises a plant extract of Theobroma cacao as an active component. The article, beverage and food for suppressing infection of influenza are prepared by adsorption of, impregnation with or addition of the anti-influenza virus agent. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

The present invention relates to an anti-influenza virus agent comprising a plant-derived extract having an influenza virus infection-suppressing action as an active ingredient, an influenza infection-suppressing article and food and drink obtained by adsorbing, impregnating and adding it .

  The influenza virus that causes the influenza epidemic is an RNA virus having an envelope membrane with a diameter of about 1 / 10,000 mm in diameter every year. Although it is classified into three types, A, B, and C, due to the difference in antigenicity, it is A type and B type that show a trendy spread. Two types of glycoproteins, haemagglutinin (HA) and neuraminidase (NA), protrude in the form of spikes on the surface of these virus particles, and gene RNA segmented into 8 segments exists inside. HA and NA on the surface of the virus frequently mutate within the same subtype, and new antigenic variants appear every year.

  Influenza virus released by droplets due to cough enters the human nose and mouth, adsorbs to the mucosal epithelial cells of the upper respiratory tract by the spike-like glycoprotein HA on the surface of the virus, and starts to proliferate after entering the cells. Recent research has revealed the mechanism of viral infection. Viruses bind to receptors consisting of sugar chains on the surface layer of human target cells, are taken into endosomes, enter cells by fusion of viral membranes and endosomal membranes, and express and replicate viral genes, and from host cell membranes. Progeny virus particles are formed and propagated by budding.

  Infection with influenza virus causes symptoms such as sudden fever, headache, joint pain, general malaise in a few days, followed by respiratory symptoms such as cough, sore throat, runny nose and stuffy nose. Unlike the so-called cold, it is highly infectious and causes an explosive epidemic in a short period of time. In addition, the structure of the HA protein of influenza virus repeats mutations every year, and the fact that antibodies made by past infections are not very useful is also a factor that spreads infection.

  In order to suppress infection with influenza virus, inhibition of adsorption to epithelial cells, inhibition of entry into cells, inhibition of gene transcription / replication, inhibition of protein synthesis, inhibition of release from cells, etc. are considered. Each is a target for antiviral drugs. To date, antiviral drugs such as amantadine, rimantadine, and zanamivir have been developed, but side effects such as hypersensitivity, neuropsychiatric symptoms, digestive system symptoms, and autonomic nervous system symptoms have been reported. Caution must be taken.

  In addition, influenza viruses are thought to be difficult to control by vaccination because influenza viruses infect and propagate in the respiratory mucosal epithelium and the epidemic type of the year cannot be accurately predicted. Frequent gargling, avoiding throat dryness, and adequate nutrition and rest are currently considered the most effective preventive measures. Development of an anti-influenza virus agent that is highly effective in suppressing infection and has no safety problems and can be used on a daily basis is desired.

  In recent years, polyphenol components of tea and black tea have been reported as anti-influenza materials derived from natural products (see, for example, Non-Patent Documents 1 and 2), and gargle of black tea suppresses actual viral infection by human tests. (For example, refer nonpatent literature 3). In addition, it has been reported that a flavonoid component derived from urgonum exhibits an influenza infection suppression effect due to a viral sialidase inhibitory activity (see, for example, Non-Patent Document 4). Furthermore, Katsushida Futoshi Soichito (for example, refer to Patent Document 1), Kurofu currant extract (for example, refer to Patent Document 2), potato anthocyanin pigment (for example, refer to Patent Document 3), Antiviral effects such as guava leaf extract (for example, see Patent Document 4) and rabu hemp extract (for example, see Patent Document 5) have been reported.

  Moreover, in the Rosaceae plant, an anti-influenza agent containing the flower bud or petal extract as an active ingredient has been disclosed (for example, see Patent Document 6), but the anti-influenza virus effect of karin has been clarified. Absent. In the genus Citrus, the preventive effect of the seed component against infection by bacteria and viruses is disclosed (for example, see Patent Document 7), but the effect of Citrus leaves is not mentioned. That is, there is no report on the influenza virus infection inhibitory effect of the plant extract shown in this patent, and it has been clarified for the first time by this patent.

Journal of Infectious Diseases, 68 (7) 824-829 (1994) Journal of Infectious Diseases, 70 (11) 1190-1192 (1996) Journal of Infectious Diseases, 71 (6) 487-494 (1997) Chem. Pharm. Bull. 38 (5) 1329-1332 (1990)

JP-A-6-199680 JP 2000-212092 A JP 2001-316399 A JP 2000-273048 A JP-A-11-71296 JP 2002-145790 A Special table 2002-020305 gazette

The object of the present invention is to use a highly safe plant extract that can be used with peace of mind on a daily basis, exhibiting a high inhibitory effect against influenza virus infection, and having no side effects , adsorbing and impregnating it It is providing the influenza infection suppression article and food and drink which are made to add .

In order to solve the above problems, the present inventors have focused on plants, herbal medicines, herbs, etc. that have been used for foods, fragrances, teas, etc., since they have no side effects and are highly safe. In order to find a plant extract having an action, a test was carried out using the inhibitory effect of influenza virus (type A virus strain: Udonn / 307/72 H3N2 type B virus: Johannesburg / 5/99) on MDCK cells as an index. . As a result, extract obtained Ri by cocoa is found to have an infection inhibiting effect of strong influenza virus, and completed the present invention product.

That is, the present invention is an anti-influenza virus agent characterized by containing, as an active ingredient, plant extract cocoa (Theobroma cacao).

Moreover, this invention is an influenza infection suppression article and food-drinks which adsorb | suck, impregnate, and add the said anti-influenza virus agent.

The present invention provides an anti-influenza virus agent having a highly safe plant extract as an active ingredient and having a strong infection-suppressing action against influenza virus. Furthermore, plant extracts as an active ingredient of the anti-influenza virus agent of the present invention because of its high safety masks, air filters, clothing, wet tissues, adsorbed to a spray solution or the like, impregnated, by adding, influenza It can be widely used in daily life as a virus infection suppression product. Furthermore, it can be added to foods and drinks such as chewing gum, candy, tablet confectionery, beverages, etc., and can be used and taken on a daily basis as foods and drinks having an anti-influenza virus action. The product of the present invention is effective for prevention of influenza virus infection and alleviation of disease symptoms caused by influenza virus.

In the roux which is the raw material of the product of the present invention, the whole can be used, but for citrus plants , leaves are used for karin, its fruits are used for everlasting and linden, flowers are used for cats claw and bayberries. It is desirable to use seeds for bark, allspice and cacao. As for cacao seeds, those that have been roasted can also be used.

  The method for obtaining the extract of the present invention from the pulverized plant is not particularly limited, but water, methanol, ethanol, n-propanol, and lower alcohols such as n-butanol, ether, ethyl acetate, acetone, glycerin, propylene glycol. One or two or more mixed solvents of organic solvents such as these are added, and extraction is performed by a conventional extraction method. However, in consideration of taking the anti-influenza virus agent of the present invention orally, it is desirable to extract using water, ethanol or a mixture thereof from the viewpoint of safety.

  The extraction condition is not particularly limited, but is preferably about 1 to 5 hours at 50 to 90 ° C. The extract can be filtered and the extraction solvent can be distilled off, followed by concentration or lyophilization under reduced pressure. Further, those obtained by fractionating and purifying these extracts by organic solvent fractionation, column chromatography or the like can also be used.

  The form of use of the product of the present invention is not particularly limited, and powders, tablets, troches can be obtained by adding a solvent, a dispersant, a pharmaceutical carrier, an emulsifier, a diluent, a stabilizer, etc. to the plant extract exemplified as an active ingredient. Preparations, inhalants, mouthwashes, mouthwashes, suppositories, injections, etc., and can be prepared as an oral route, airway administration, intravenous administration, rectal administration, subcutaneous administration, skin Internal administration etc. can be illustrated. In this case, the dose to adults is preferably 10 to 2000 mg / day for each extract, but is not limited to this value. The amount of the extract added to various preparations varies depending on the form of the preparation, but it is preferable to add the extract so that the ratio is 0.001 wt% or more, preferably about 0.01 wt% or more.

  Moreover, the infection control article which can contribute to influenza prevention can be provided by adsorb | sucking, impregnating, and adding the anti-influenza virus agent of this invention to a mask, an air-conditioner filter, clothing, wet tissue, a spray liquid, etc. The amount of adsorbed and added plant extract in these uses varies depending on the form of the infection-suppressing product, and cannot be specified in general, but is added so that the proportion is 0.001 to 5% by weight. Is preferred.

  In addition, since the anti-influenza virus agent of the present invention has high safety, for example, chewing gum, candy, tablet confectionery, gummy jelly, chocolate, biscuits and other confectionery, ice cream, sorbet and other frozen confectionery, beverages, soup, jam and other food and drink And can be used on a daily basis. The amount added varies depending on the form of use and the taste of the extract, but it is added in an amount of 0.001 to 5% by weight, preferably about 0.01 to 1% by weight, based on the food or drink. Is preferred.

  Hereinafter, the present invention will be specifically described with reference to examples and test examples, but the present invention is not limited thereto.

  A reflux condenser is attached to a flask obtained by adding 300 ml of 50% ethanol to 30 g of Linden flower powder, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered off, the solvent was removed, and the residue was freeze-dried to obtain 4.86 g of the extract.

In the same manner, extracts of citrus leaves, cat's claw bark, bayberry bark, roux whole grass, everlasting flowers, allspice seeds, cacao seeds (roasted / not roasted), and quince fruits using 50% ethanol. The extract was prepared by concentrating and lyophilizing the extract. In addition, about the cacao seed (roast, non-roast), what grind | pulverized with 10 times amount n-hexane was used after grind | pulverizing. The yield of each extract is shown in Table 1.

A reflux condenser is attached to a flask in which 300 ml of 70% ethanol is added to 30 g of Citrus plant leaf powder, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered off, the solvent was removed, and freeze-dried to obtain 4.68 g of the extract.

  Similarly, bayberry bark was extracted with 70% ethanol, and the extract was concentrated and lyophilized to prepare an extract. The yield of each extract is shown in Table 1.

  A reflux condenser is attached to a flask in which 300 ml of 100% ethanol is added to 30 g of cat's claw bark powder, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered off, the solvent was removed, and the residue was dried to obtain 3.42 g of the extract.

Similarly, extract citrus leaves, everlasting flowers, linden flowers, allspice seeds, cacao seeds (roasted / not roasted), and carin seeds using 100% ethanol, and concentrate and dry the extract. An extract was prepared. In addition, about the cacao seed (roast, non-roast), what grind | pulverized with 10 times amount n-hexane was used after grind | pulverizing. The yield of each extract is shown in Table 1.

  A reflux condenser is attached to a flask in which 300 ml of water is added to 30 g of bayberry bark powder, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered off, the solvent was removed, and the residue was freeze-dried to obtain 6.78 g of the extract.

In the same manner, extract extracts of citrus leaves, cat's claw bark, whole roux, everlasting flowers, linden flowers, and allspice seeds using water, and concentrate and freeze-dry the extract to prepare the extract. did. The yield of each extract is shown in Table 1.

  A reflux condenser is attached to a flask in which 300 ml of 100% methanol is added to 30 g of Everlasting flower powder, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered off, the solvent was removed, and the residue was dried to obtain 2.49 g of the extract. The yield of the extract is shown in Table 1.

  A reflux condenser is attached to a flask in which 300 ml of ethyl acetate is added to 30 g of bayberry bark powder, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered off, the solvent was removed, and the residue was dried to obtain 9.27 g of the extract.

Similarly, extracts of Citrus plant leaves were extracted with ethyl acetate, and the extract was concentrated and dried to prepare an extract. The yield of each extract is shown in Table 1.

A reflux condenser is attached to a flask obtained by adding 300 ml of acetone to 30 g of Citrus plant leaf powder, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered off, the solvent was removed, and the residue was dried to obtain 2.37 g of the extract. The yield of the extract is shown in Table 1.

  A reflux condenser is attached to a flask in which 300 ml of n-butanol is added to 30 g of roux whole plant powder, and extraction is performed while refluxing for 1 hour. The obtained extract was filtered off, the solvent was removed, and the residue was dried to obtain 5.82 g of the extract.

  Similarly, catscrow bark was extracted with n-butanol, and the extract was concentrated and dried to prepare an extract. The yield of each extract is shown in Table 1.

Test example 1

Using the extract, which is the product of the present invention shown in Examples 1 to 8, as a sample, the influenza virus infection inhibitory effect was examined. As influenza viruses, A / Udorn / 307/72 (H3N2) and B / Johannesburg / 5 / 99N2 strains were used. A sample stock solution was prepared by dissolving a sample in dimethyl sulfoxide (50 mg / ml). The sample stock solution Tris-Glucose-Saline (TGS) at ten-fold serial dilutions (10 to ⁶ fold dilution), these sample diluent and virus solution (1000PFU (plaque forming unit) / ml) 1: mixture 1 And reacted at room temperature for 30 minutes. 0.1 ml of this was inoculated into a monolayer culture (35 mm diameter petri dish) of Madin-Darby canine kidney (MDCK) cells, the virus was adsorbed at room temperature for 1 hour, and 2 ml of L-15 agarose medium was poured and solidified at 34 ° C. After culturing for 3 days, the number of plaques produced was counted. The dilution multiple of the sample where the number of plaques was 50% or less of the TGS-treated control was defined as the infection neutralization value. The influenza virus infection inhibitory activity (infection neutralization value) of the extracts of the present invention prepared in Examples 1 to 8 is shown in Table 2 below. All of these extracts, which are the products of the present invention, had an infection neutralization value of 10 ³ or more, and showed a strong virus infection suppression effect.

  Using the plant extracts prepared in Examples 1-8, mouthwash, inhalant, troche, spray, chewing gum, candy, tablet confectionery, beverage, powder, tablet, gargle, gummy jelly, chocolate, biscuit Ice, sorbet, soup, jam, wet tissue, and mask were prepared. The prescription was shown as an Example below.

Mouthwash prescription <br/> Ethanol 2.0% by weight
Fragrance 1.0% by weight
Saccharin 0.05 wt%
Chlorhexidine hydrochloride 0.01% by weight
Bayberry bark 70% ethanol extract (Example 2) 0.5 wt%
Water remaining
100.0% by weight

Inhalation formula <br/> Ethanol 5.0% by weight
Everlasting flower 100% ethanol extract (Example 3) 1.0 wt%
Water remaining
100.0% by weight

Lozenge formulation Glucose 72.3 wt%
Lactose 19.0% by weight
Gum arabic 6.0% by weight
Fragrance 1.0% by weight
Sodium monofluorophosphate 0.7% by weight
Cat's claw bark
n-Butanol extract (Example 8) 1.0 wt%
100.0% by weight

Formulation of spray liquid Ethanol 25.0% by weight
Citric acid 1.5% by weight
Trisodium citrate 1.0 wt%
Roux whole plant 50% ethanol extract (Example 1) 0.5 wt%
Water remaining
100.0% by weight

Chewing gum formula Gum base 20.0% by weight
54.7% by weight sugar
Glucose 15.0% by weight
Minamata 9.3 wt%
Fragrance 0.5% by weight
Cocoa seed (roasted)
50% ethanol extract (Example 1) 0.2% by weight
Citrus plant leaves 50% ethanol extract (Example 1) 0.3 wt%
100.0% by weight

Candy formula <br/> Sugar 50.0% by weight
Minamata 34.0 wt%
Citric acid 1.0% by weight
Fragrance 0.2 wt%
Linden flower water extract (Example 4) 0.4 wt%
Water remaining
100.0% by weight

Formula for tablet confectionery Sugar 76.1% by weight
Glucose 19.0% by weight
Sucrose fatty acid ester 0.2% by weight
Fragrance 0.2 wt%
Allspice seed 50% ethanol extract (Example 1) 0.5% by weight
Water remaining
100.0% by weight

Beverage prescription
Citrus fruit juice 30.00 wt%
Isomerized sugar 15.24% by weight
Citric acid 0.10% by weight
Vitamin C 0.04% by weight
Perfume 0.10% by weight
Carin 50% ethanol extract (Example 1) 0.10 wt%
Water remaining
100.00% by weight

Formulation of powder <br/> Corn starch 55.0% by weight
Carboxycellulose 40.0% by weight
Everlasting Flower Water Extract (Example 4) 5.0 wt%
100.0% by weight

Tablet formulation Lactose 70.0% by weight
Crystalline cellulose 15.0% by weight
Magnesium stearate 5.0 wt%
Linden flower 50% ethanol extract (Example 1) 10.0 wt%
100.0% by weight

Formula of mouthwash <br/> Ethanol 2.00% by weight
Perfume 1.00% by weight
Saccharin 0.05 wt%
Chlorhexidine hydrochloride 0.01% by weight
Bayberry bark ethyl acetate extract (Example 6) 0.50 wt%
Water remaining
100.00% by weight

Gummy jelly formula <br/> Gelatin 60.00 wt%
Minamata 23.00 wt%
Sugar 8.50% by weight
Vegetable oil and fat 4.50% by weight
Mannitol 2.95% by weight
Lemon juice 1.00% by weight
Allspice seeds
100% ethanol extract (Example 3) 0.05 wt%
100.00% by weight

Chocolate formula Powdered sugar 40.80% by weight
Cocoa bitter 20.00 wt%
Whole milk powder 20.00% by weight
Cocoa butter 17.00 wt%
Mannitol 1.00% by weight
Fragrance 0.20% by weight
Citrus plant leaf water extract (Example 4) 1.00 wt%
100.00% by weight

Biscuit prescription <br/> 1st grade, low strength 25.59% by weight
Medium strength 1st class 22.22% by weight
Refined sugar 4.80% by weight
Salt 0.73 wt%
Glucose 0.78% by weight
Palm shortening 11.78 wt%
Sodium bicarbonate 0.17% by weight
Sodium bisulfite 0.16% by weight
Rice flour 1.45% by weight
Whole milk powder 1.16% by weight
Substitute milk powder 0.29 wt%
Bayberry bark 70% ethanol extract (Example 2) 0.50 wt%
Water remaining
100.00% by weight

Ice formula <br/> Nonfat dry milk 50.0% by weight
Fresh cream 25.0% by weight
Sugar 10.0% by weight
Egg yolk 10.0% by weight
Cat's Claw bark 50% ethanol extract (Example 1) 1.0 wt%
Fragrance 0.1% by weight
Water remaining
100.0% by weight

Sherbet prescription
Citrus fruit juice 25.0% by weight
Sugar 25.0% by weight
Egg white 10.0% by weight
Citrus plant leaf water extract (Example 4) 2.0% by weight
Water remaining
100.0% by weight

Soup formula <br/> Milk 60.00 wt%
Onion 20.00% by weight
Carrot 10.00% by weight
Vegetable bouillon 1.00% by weight
0.10% by weight of butter
Pepper 0.05 wt%
0.05% salt by weight
Allspice seed water extract (Example 4) 1.00% by weight
Water remaining
100.00% by weight

Jam prescription <br/> Pulp 4.0% by weight
65.0% by weight sugar
Kiyosumi juice 25.0% by weight
Citric acid 0.5% by weight
Carin 50% ethanol extract (Example 1) 2.0% by weight
Water remaining
100.0% by weight

Prescription for wet tissue <br/> Medium solution Benzalkonium chloride 0.2% by weight
Whole roux 50% ethanol extract (Example 1) 0.2 wt%
Water remaining
100.0% by weight
20 pieces of nonwoven fabric (150 × 200 mm) were stacked, and the above components were impregnated with 3 times the weight of the nonwoven fabric.

Mask formulation Linden flower 100% ethanol extract (Example 3) 1.0 wt%
Water remaining
100.0% by weight
The nonwoven fabric was dipped in the above components, lightly dehydrated and dried.

Claims (3)

Anti-influenza virus agent characterized by a plant extract of cacao (Theobroma cacao) as an active ingredient. A product for suppressing influenza infection comprising adsorbing, impregnating and adding the anti-influenza virus agent according to claim 1.   A food or drink comprising the anti-influenza virus agent according to claim 1.