WO2006115123A1

WO2006115123A1 - Virus-inactivating agent

Info

Publication number: WO2006115123A1
Application number: PCT/JP2006/308128
Authority: WO
Inventors: Kimitoshi Noda; Naoko Morinaga; Motoyuki Tagashira
Original assignee: Asahi Breweries, Ltd.
Priority date: 2005-04-19
Filing date: 2006-04-18
Publication date: 2006-11-02
Also published as: JPWO2006115123A1

Abstract

A virus-inactivating agent comprising proanthocyanidin polyphenol, especially preferably a polyphenol obtained from an apple immature fruit or a hop bract, having an inactivating (infection prevention) potency against viruses, such as influenza virus. In the use of this virus-inactivating agent, the inactivating (infection prevention) effect against influenza virus is efficaciously exerted by the action of polyphenol derived from or contained in apples or polyphenol derived from or contained in hops.

Description

Specification

Virus inactivating agent

Technical field

[0001] The present invention relates to a virus inactivating agent, which is a porcine anthocyanin-based polyphenol having an inactivating effect on a virus such as influenza virus, particularly preferably a polyphenol derived from an immature apple or hop koji. .

This application claims priority based on Japanese Patent Application No. 2005-121343 filed in Japan on April 19, 2005, the contents of which are incorporated herein by reference.

Background art

[0002] The power of research on natural substances having antiviral activity so far includes proanthocyanidin polymers (see Patent Document 1). The polymer is a proanthocyanidin polymer having 2 to 30 flavonoid units that can also have a plant power of croton-recleri and is said to be effective in the treatment of respiratory virus infections such as influenza. Specifically, this document describes the therapeutic effect of broanthocyanine polymer A (having a degree of polymerization of 2 to 11 flavonoid units and an average of about 7) against influenza virus A virus.

In addition, there is an influenza virus infection preventive agent containing tea polyphenol as an active ingredient (see Patent Document 2). In this document, tea catechins such as epicarocatechin gallate are listed as tea polyphenols.

[0003] Patent Document 1: Patent No. 3448052

Patent Document 2: JP-A-3-101623

Disclosure of the invention

Problems to be solved by the invention

[0004] The proanthocyanidin polymer described in the above-mentioned Patent Document 1 is effective for the treatment of respiratory viral infections, but the raw material is a croton-type tree that grows in a specific environment such as Amazon. Therefore, it is not suitable for simple and mass production. In recent years, the removal of such wood has been severely restricted for the purpose of protecting genetic resources. further Easy take-off and breeding of non-native species other than the original habitat may disrupt the ecosystem.

In other words, if it is possible to provide a polyphenol having antiviral activity that is more effective than conventional materials and can be obtained more easily and cheaply than croton species, it can be used as industrially valuable. Be expected.

Means for solving the problem

[0005] As a result of intensive studies on the above problems, the present inventors have found that they have an effect on proanthocyanin-based polyphenola influenza viruses derived from or containing hop koji or apple immature fruit. Furthermore, it was confirmed that these active substances have a stronger activity than tea catechin, and the present invention has been completed.

The term “polyphenol” as used herein is a general term for compounds having a plurality of phenolic hydroxyl groups in a molecule mainly contained in a plant body. “Proanthocyanin-based polyphenol” is a red pigment by hydrolysis. A compound that produces an anthocyanin (cyanidine, delphidin or pelargine)! Uh.

That is, the first of the present invention is to provide a virus inactivating agent containing a proanthocyanidin-based polyphenol as an active ingredient that inactivates viruses. The second of the present invention is to provide a virus inactivating agent in which the proanthocyanin-based polyphenol is a polyphenol obtained from an apple, particularly an unripe fruit of an apple. The third of the present invention is to provide a virus inactivating agent, wherein the proanthocyanidin-based polyphenol is a polyphenol from which hops, in particular, hop cake is also obtained.

Here, in the virus inactivating agent of the present invention, the Puroantoshia - Gin systems the preferred polyphenol 5-95 ^_0/0, more preferably 20 to 95 weight ^_0/0, it is preferable to contain. In particular, Puroantoshia - gin based polyphenol is also (from or contained in apples) obtained Ringoka when a polyphenol is preferably a virus inactivating agent is 20 to 80 wt%, more preferably 50 to 70 weight ⁰ / ₀ , containing the polyphenol. In addition, when the proanthocyanidin-based polyphenol is a polyphenol that also has hopping power (derived from or contained in hops), it is preferably 20 to 70% by weight, more preferably 25 to 55% by weight as a virus inactivating agent %, The polyphenol is contained. [0007] Further, in the present invention, the proanthocyanidin-based polyphenol alcohol may be a proanthocyanine that does not permeate the membrane when treated with an ultrafiltration membrane having a molecular weight cut off of 1,000 or more. preferable.

Furthermore, the present invention provides a beverage or food product containing the above-mentioned virus inactivating agent, preferably in a final concentration range of 1 to 10, OOOppm.

In addition, the present invention also provides a therapeutic method in which, together with the above-mentioned medicine containing the virus inactivating agent, the medicine is administered as an oral or parenteral agent to a patient in a total daily dose of 2 mg to LOOO mg. To do.

The polyphenol provided by the present invention is stronger than the tea catechin described in JP-A-3-101623 in the strength of the virus growth inhibitory activity, and is applied to beverages and foods. It is excellent also in the taste which becomes important when doing.

The invention's effect

[0008] Since the virus inactivating agent of the present invention has anti-influenza virus activity, it is effective against influenza virus. In addition, since the components that inactivate influenza virus are hops, especially hop koji, or polyphenols derived from or containing apples, especially apple immature fruits, there is an advantage that raw materials can be easily obtained.

Brief Description of Drawings

[0009] FIG. 1 is a diagram in which the degree of growth of Sindbis virus in Example 10 is evaluated by viability of Vero cells. The higher the value on the vertical axis, the more cells are alive.

FIG. 2 shows the degree of proliferation of influenza virus in Example 11 evaluated by the hemagglutination activity of chorioallantoic fluid. The vertical axis shows the maximum dilution factor showing hemagglutination activity, and the higher this value, the more influenza virus grows! /.

FIG. 3 is a graph showing an evaluation of the hemagglutination activity of chickens possessed by influenza virus in Example 12. The vertical axis is a score of the degree of erythrocyte aggregation. The higher this value, the stronger the infectivity.

BEST MODE FOR CARRYING OUT THE INVENTION

[0010] The unripe apple fruit used as the raw material of the present invention refers to an fruit that has been artificially picked before the fruit of the apple has matured, or that has fallen from the apple fruit tree naturally by dropping or the like. The hop koji used as the raw material of the present invention is obtained by removing the rubulin portion from the hop koji, and in general, the hop koji is obtained by pulverizing the hop koji and removing the rubulin portion by sieving. However, in recent beer brewing, it is useful for beer brewing in order to save the trouble of hopping and removing the hop cake. It tends to be used for brewing beer as hop pellets. Accordingly, the raw material of the present invention is not particularly limited as long as it contains hop cake, and there is no problem even if hop fruit and hop pellets containing hop cake are used as raw materials.

[0011] These raw materials are extracted by squeezing with a juice or an aqueous alcohol solution. The extracted liquid is an extract containing a virus inactive ingredient. Further, if necessary, the virus-inactive component can be purified by using a column packed with granular resin having affinity for polyphenol, etc., and the degree of purification can be increased. However, this process is only to increase the degree of purification of the virus inactivating agent, and can be omitted if not necessary. The virus inactive ingredient is a granular resin having affinity for polyphenol (hereinafter “ In the purification process using a column packed with “fat” (t), etc. (hereinafter “column” ヽぅ), the extract containing the virus inactivating component is passed through the column and the column is thoroughly washed. Thereafter, the virus inactivating agent adsorbed on the column may be eluted, or granular rosin may be immersed in an extract containing a virus inactivating component and batch-treated.

[0012] When adsorbing the virus inactivating agent on the sardine, if necessary, after cooling the extract containing the virus inactivating component to about 5 to 30 ° C in order to increase the adsorption efficiency, It is desirable to reduce the concentration of the organic solvent in the extract beforehand by concentration under reduced pressure. As the material of the resin, hydroxypropylated dextran, hydrophilic bull polymer, styrene-dibutylbenzene polymer and the like can be used.

Subsequently, the scab can be washed to increase the degree of purification of the virus inactivating agent. As a solvent used for washing, water or an aqueous ethanol solution of 1 to 10 wZw% (weight Z weight%) is suitable, and it is desirable to wash using a solvent amount of about 1 to 10 times the amount of greaves.

[0013] Subsequently, the virus inactivating agent is desorbed and eluted from the sorbate adsorbed with polyphenols. Melting As the solvent to be used, hydrous alcohol, hydrous acetone, hydrous acetonitrile or the like can be used. Particularly suitable examples include ethanol aqueous solution or ethanol of 10 wZw% or more. Desirably, the amount of elution solvent to be passed is about 2 to 6 times the amount of sallow. The virus inactivating agent can be obtained as a powder by removing the solvent from the obtained eluate by a conventional method such as concentration, freeze-drying or spray-drying. Also, alcohol, acetone, acetonitrile, etc. can be recovered and reused during concentration under reduced pressure. The synthetic resin used can be repeatedly used after being washed with an alcohol aqueous solution of 80 vZv% (capacity Z volume%) or more, an aqueous sodium hydroxide solution of about 0.05 N, and the like.

The virus inactivating agent obtained by force can be put to practical use as it is, but the degree of purification can be further increased by a method using an ultrafiltration membrane as described below. However, this process is only for increasing the purity of the virus inactivating agent and can be omitted if not necessary.

[0014] The step of purifying the virus inactive ingredient using an ultrafiltration membrane is performed by ultrafiltration with a molecular weight cut off of 1,000 or more from the solution containing the virus inactivating agent obtained by the above method. Treat with membrane. The solution is preferably water or a solution of an organic solvent miscible with water. A suitable example is a 5-50% aqueous ethanol solution. The membrane material can be used without particular limitation as long as it is usually used as a material for an ultrafiltration membrane, such as cell mouth, cellulose acetate, polysanolefon, polypropylene, polyester, polyethersulfone, PVDF. In addition, if the molecular weight cut off is 1,000 or more, it can be used without any problem. However, if a membrane with a large molecular weight cut off is used, the yield will be drastically reduced. Since the time required for the treatment becomes long, an ultrafiltration membrane having a fractional molecular weight of about 3,000 to 50,000 is preferred. In addition, it is desirable that the treatment is performed until the amount of the remaining liquid is about iZio to iZioo at the start of the treatment, depending on the type of the extraction solvent and the ratio of the extraction solvent and hops or hops. The pressure at that time is preferably about 0.1 to LO. OkgZcm ^{2 although} it depends on the ultrafiltration membrane and the filtration device. If necessary, after the treatment, the remaining liquid can be diluted again with a suitable solvent such as water and retreated in the same manner to increase the degree of purification.

[0015] The solvent of the obtained upper liquid residue is concentrated by a usual method such as concentration, freeze drying, spray drying, or the like. Except, the virus inactivating agent can be obtained as a powder. Also, alcohol, acetone, acetonitrile, etc. can be recovered and reused during concentration under reduced pressure.

The virus inactivating agent thus obtained is an odorless skin-colored, brown or light yellow powder with a faint bitter taste, adsorbed on a synthetic resin having an affinity for polyphenol, and a molecular weight cut off. Proanthocyanidins that do not permeate the membrane when treated with more than 1,000 ultrafiltration membranes.

When the raw material is hop koji, the yield is 0.5-20. Ow / w% in terms of hop koji weight, and 0.5-15. OwZw% in terms of hop koji weight. In the case of fruit, it is 0.5 to 15. OwZw% in terms of the weight of immature apple.

[0016] The content (rate) of the active ingredient, the proanthocyanin-based polyphenol, in the obtained virus inactivating agent varies depending on the difference in the purification process described above, and the content is small. However, depending on the amount of intake and dose, etc., a sufficient effect can be expected, and there is no limitation. More specifically, if proanthocyanidin-based polyphenols are contained in the virus inactivator in an amount of 2% by weight or more, the effect is exhibited as an active ingredient. In consideration of time and labor, it is preferably contained in the range of 5 to 95% by weight, more preferably 20 to 95% by weight.

In particular, when the proanthocyanin-based polyphenol is a polyphenol obtained from apple strength, the virus inactivating agent preferably contains 20 to 80% by weight, more preferably 50 to 70% by weight. In addition, when the proanthocyanin-based polyphenol is a polyphenol obtained with a hopping force, the virus inactivating agent is preferably 20 to 70% by weight, more preferably 25 to 55% by weight. contains.

[0017] The content (% by weight) of the proanthocyanin-based polyphenol in the virus inactivating agent is generally determined by the method of Porter et al. (Porter et al., Phytochemistry 25, 223-230 (1986)). This method is a method in which proanthosidin-based polyphenol-specific interflavan bonds are hydrolyzed in the presence of alcohol, and the resulting red pigments such as pelargosine, cyadin, and delphidinidine are quantified at an absorbance of 550 nm. It is. Proanthocyanidin-based polyphenols contain a variety of substances, so the light absorption usually obtained The degree can be converted to the absorbance of a relatively simple structure such as procyan-zine B1, such as procyan-zine B1, and expressed as the content of procyan-zine B1.

Therefore, by using the method of Porter et al., The content of proanthocyanidin-based polyphenol in the virus inactivating agent can be determined.

[0018] The virus inactivating agent obtained by force can be formulated with commonly used carriers, auxiliaries, additives, etc., and as a pharmaceutical product as an oral or parenteral product according to a conventional method. It can be used and mixed with food ingredients to make food and drink.

Pharmaceutical products include tablets, capsules, powders, granules, and syrups as oral preparations, and non-oral preparations such as ointments, creams, and liquids, and injections such as sterile solutions and suspensions. is there. It can also be made into mouth preparations such as mouthwash. When these products are administered to the human body as medicines, 2 mg to 500 mg is administered once or several times a day, that is, 2 mg to: LOOO mg daily. By carrying out this treatment method, the inactive effect against viruses such as influenza virus can be sufficiently achieved.

[0019] The pharmaceutical containing the virus inactivating agent of the present invention is a physiologically recognized vehicle or carrier.

It can take the required unit volume form with excipients, integrants, stabilizers, flavors, and the like. The adjuncts mixed in tablets and capsules are as follows. Binders such as tragacanth, gum arabic, corn starch, gelatin, excipients such as microcrystalline cellulose, corn starch, total gelatinized starch, swelling agents such as alginic acid, lubricants such as magnesium stearate , Sweeteners such as sucrose, lactose, saccharin, peppermint, hard mono oil, flavoring such as cherry. In the case of capsules, the above materials can further contain a liquid carrier such as fats and oils, and other materials can be used as coating agents or the physical form of the preparation can be changed by other methods. be able to. For example, tablets can be coated with shellac and sugar. Syrup or elixir is sucrose as a sweetener

As preservatives, methyl or propyl parabens, pigments and flavoring agents such as cherry or orange flavors can be included.

When the pharmaceutical form is an oral preparation such as a tablet, capsule, powder, granule or the like, the content of the virus inactivating agent is preferably 3 to 50% by weight, more preferably 5 to 30% by weight. Range.

[0020] A sterile composition for injection comprises an active substance in a vehicle such as water for injection, a naturally occurring vegetable oil such as sesame oil, coconut oil, peanut oil, cottonseed oil, or a synthetic fat vehicle such as ethylolate. It can be formulated by conventional methods of dissolving or suspending. Moreover, a buffering agent, a preservative, an anti-oxidation agent, etc. can be mix | blended as needed. As an external preparation, vaseline, greaffin, oils and fats, lanolin, macrogol or the like is used as a base material, and an ointment, cream or the like can be obtained by an ordinary method.

When the pharmaceutical form is an injection, oral preparation, etc., the content of the virus inactivating agent is preferably in the range of 0.2 to 10% by weight, more preferably 0.5 to 5% by weight.

[0021] The beverage or food product containing the virus inactivating agent of the present invention may be in the form of the above-mentioned preparation, but in the form of candy, rice cracker, cookie, beverage, etc., the required amount is added to each raw material, It can also be processed and manufactured by a general manufacturing method. Ingestion as health food and functional food is used for disease prevention and health maintenance, so it can be divided into several times a day for oral consumption and taken as a daily product containing 5 mg to 500 mg. .

When a virus inactivating agent is added to these beverages or foods, the virus inactivating agent may be added as a powder, but preferably the virus inactivating agent is added in an aqueous solution of 1 to 2% by weight or It is desirable to add an aqueous alcohol solution or an alcohol solution so that the final concentration is usually 1 to 10, OOOppm, preferably 100 to 5000 ppm, for beverages or foods.

Hereinafter, although an Example is shown, this invention is not limited to this.

Example 1

[0022] (Preparation of virus inactivating agent from unripe apple fruit)

400g of apple immature fruit (average weight 5.03g) was homogenized with 1% hydrochloric acid methanol and extracted with heating under reflux (3 times). The extract was concentrated under reduced pressure to distill off the methanol, In addition, it was distributed (twice), and the aqueous layer was collected. After filtration, it was diluted to 200 ml with distilled water. Furthermore, it was purified by solid phase extraction using Sep-pak C18 and freeze-dried to obtain 8.9 g of a virus inactivating agent. The yield of immature apple fruit was 2.2%.

In addition, this virus inactivating agent was added to size exclusion chromatography (Kurumatani et al., J. Liq. Chromatogr. Realt. Technol. 28, 1971-1983, (2005)), the average molecular weight of the virus inactivating agent was calculated to be about 1,500. This is equivalent to about a pentamer in terms of catechin.

When this virus inactivating agent was quantified by proanthocyanin using the Porter method, it was found to be 63% in terms of procyanidin B1.

Example 2

[0023] (Preparation of virus inactivating agent from hop cake)

50 g of hop koji was extracted with 1,000 ml of 40% ethanol aqueous solution with stirring at 50 ° C. for 60 minutes. After filtration, the solution was concentrated under reduced pressure until the volume reached 500 ml. The concentrated solution was passed through a column packed with 150 ml of styrene-dibulene benzoate (Separbeads 70 manufactured by Mitsubishi Chemical Co., Ltd.), and then with 500 ml of water. Washed. Further, 600 ml of 50% ethanol aqueous solution was passed through the same column, and the same eluate was collected and lyophilized to obtain 1.7 g of a virus inactive ingredient as a light yellow powder with an odorless faint bitterness. It was. The yield of hop repulsion was 3.4%.

When this virus inactivating agent was quantified with proanthocyanin using the Porter method, it was 36% in terms of procyanin-B1.

Example 3

[0024] (Further purification of virus inactivating agent using ultrafiltration membrane)

5 g of the virus inactivating component obtained in the same manner as in Example 2 was dissolved in 500 ml of a 20% aqueous ethanol solution and treated with an ultrafiltration membrane (YM10 manufactured by Millipore) having a molecular weight cut-off of 10,000. The remaining liquid that did not pass through this membrane was concentrated and lyophilized to obtain 2.2 g of a further purified virus inactivating agent as a yellow to brown powder. As in Example 1, when measured using size exclusion chromatography, the average molecular weight of the virus inactivating agent was calculated to be about 6,300. This corresponds to approximately 22-mer in terms of catechin.

When this powder was subjected to HPLC analysis, it became a characteristic chromatogram, and catechin determination (a method described in the official food analysis method), which is one of the general methods for quantifying polyphenols, was performed and converted to catechin content. A value of 40.6% was obtained. (HPLC conditions) Equipment: Shimadzu LC—10A system, column: ODS-80TM (Tosohichi, 4.6mm ID X 25cm), mobile phase: (A liquid: B liquid) = (100: 0) Same force (50: 50) 30-minute linear gradient, liquid A: 5% acetonitrile Lil (containing 0.1% HC1), B solution: acetonitrile, flow rate: 1.0 ml / min, sample injection amount: 20 g, detection: multi-wavelength detection at 200-300 nm.

The official food analysis method (HPLC method) for catechin determination can be measured, for example, according to the method described in the Ministry of Agriculture, Forestry and Fisheries of Vegetables, Tea Experiment Station “Tea Analysis Method” Tea Industry Research Report No. 71 (1990).

This powder (virus inactivating agent) was quantified with proanthocyanin using the Porter method, and as a result, it was 53% in terms of procyanin B1.

Example 4

[0025] (Tablets, capsules)

Material obtained according to example 3. 10. Og

Lactose 75. Og

Magnesium stearate 15. Og

Total 100. Og

The above respective parts by weight were uniformly mixed, and tablets and capsules were prepared according to a conventional method. In addition, instead of the substance obtained according to Example 3, tablets and capsules to which the substance obtained according to Examples 1 and 2 was added were obtained in the same manner.

Example 5

[0026] (Powder, granule)

Material obtained according to Example 3. 20. Og

Starch 30. Og

Lactose 50. Og

Total 100. Og

Each said weight part was mixed uniformly, and it was set as the powder and the granule according to the conventional method. In addition, instead of the substance obtained according to Example 3, powders and granules to which the substance obtained according to Examples 1 and 2 was added were obtained in the same manner.

Example 6

[0027] (Injection)

Material obtained according to example 3. 1. Og Surfactant 9. Og

Saline 90. Og

100. Og

Each of the above weight parts was mixed by heating and sterilized to obtain an injection. Instead of the substance obtained in accordance with Example 3, powders and granules to which the substance obtained in accordance with Examples 1 and 2 was added were also obtained in the same manner.

Example 7

(飴)

Sucrose 20. Og

Minamata (75% solids) 70.0g

9.5g water

Coloring agent 0.45g

Fragrance 0. 045g

Material obtained according to Example 3 0.005 g

Total 100.0g

Using each component of each of the above parts by weight, it was made into a candy according to a conventional method. In addition, instead of the substance obtained in accordance with Example 3, soot containing the substance obtained in accordance with Examples 1 and 2, respectively, was also obtained.

Example 8

(Juice)

Concentrated tangerine juice 15.0g

Fructose 5.0 g

Chenic acid 0.2 g

Perfume 0.lg

Color 0.15g

Sodium ascorbate 0.048g

Material obtained according to Example 3 0.002 g

Water 79. 5g Total 100.0g

Using each component of the above-mentioned parts by weight, juice was prepared according to a conventional method. In addition, instead of the substance obtained according to Example 3, a juice to which the substance obtained according to Examples 1 and 2 was added was obtained in the same manner.

Example 9

(Cookies)

Soft flour 32. 0g

Whole egg 16.0g

16.0 g butter

Sand sugar 25. 0g

10.8 g of water

Baking powder 0. 198g

Material obtained according to Example 3 0.002 g

Total 100.0g

Using each component of the above-mentioned parts by weight, a cookie was prepared according to a conventional method. In addition, instead of the substance obtained according to Example 3, a cookie with the addition of the substance obtained according to Examples 1 and 2, respectively, was also obtained in the same manner.

Comparative Example 1

(Taste comparison)

Eleven panelists evaluated the taste (astringency) of the lOOppm aqueous solution of the virus inactivator or epigallocatechin gallate (commercial product: Kurita Chemical) obtained in Example 1 or Example 3 by public function. And tabulated. Panelists were asked to rate each aqueous solution on a 1-5 scale for astringency. (The astringent one was given 5 points and the weak one was given 1 point.) The results are shown in Table 1. It was confirmed that the virus inactivating agent obtained in Example 1 and Example 3 was superior in taste as compared with tea catechin.

[table 1] Score (number of people at each point) Average score

1 2 3 4 5

Example 1 2 6 3 0 0 2 .1

Example 3 4 5 2 0 0 1.8

Epigalocatechin gallate 0 0 1 8 2 4.1 Example 10

[0032] Velocity obtained by cultivating mixed solutions of various solutions of virus inactivating agent and epigallocatechin gallate (Kurita Kogyo) obtained in Examples 1 and 3 and Sindbis virus in 96-well plates. In addition to the cells, the cells were incubated for 1 hour, washed, and cultured at 37 ° C for 2 days. The concentration of the virus inactivating agent obtained in Example 1 and Example 3 was calculated based on the average molecular weight results of size exclusion chromatography with molecular weights of 1500 and 6300, respectively. After the culturing, the cell viability was evaluated by cell activity (Cell Vability) measured with a Cell counting kit (manufactured by Doujin Chemical Laboratory). As a control, cells with polyphenol and virus were used, and the cell activity indicated by the control was defined as 100%. The results are shown in Fig. 1. The virus inactivating agent obtained in Example 3 was stronger and showed a virus inactivating effect as compared with epigallocatechin gallate, which is the main component of tea catechin.

Example 11

[0033] Growth of various concentrations of the virus inactivating agent obtained in Example 3 and influenza A virus (influenza A / Beijin / 352/89 (H3N2)) at 4 ° C for 30 minutes Were inoculated into the chorioallantoic cavity and incubated at 35 ° C for 48 hours. The concentration of the virus inactivating agent of Example 3 was calculated based on the result of size exclusion chromatography with a molecular weight of 6300. After incubation, chorioallantoic fluid was collected and the amount of influenza virus was evaluated by erythrocyte aggregation activity. The result is shown in Fig.2. The virus inactivating agent of Example 3 inhibited the growth of influenza virus depending on the concentration.

Example 12

[0034] The virus inactivating agent obtained in Example 1 and Example 3 at various concentrations and A-type influenza virus (influenza A / Beijin / 352/89 (H3N2)) were mixed and ink was used at 4 ° C for 30 minutes. Then, in addition to chicken erythrocytes, the infectivity of the virus was evaluated by its aggregation activity. The concentration of the virus inactivating agent obtained in Example 1 and Example 3 was calculated with molecular weights of 1500 and 6300, respectively, from the average molecular weight results of size exclusion chromatography. Figure 3 shows the results. The virus inactivating agent obtained in Example 1 and Example 3 inhibited the hemagglutination activity of influenza A virus in a concentration-dependent manner and attenuated the infectivity of the virus.

Industrial applicability

Since the virus inactivating agent of the present invention has an effect of suppressing the growth of the virus, it has the effect of inhibiting the infection of the virus to the host, a pharmaceutical, a quasi-drug, a beverage or a food product, etc. Can be used effectively.

Claims

The scope of the claims

[I] A virus inactivating agent containing a proanthocyanidin-based polyphenol as an active ingredient that inactivates viruses.

[2] The virus inactivating agent according to claim 1, wherein the proanthocyanidin-based polyphenol is a polyphenol obtained from apple mosquito.

[3] The virus inactivating agent according to claim 2, wherein the apple is an immature apple.

[4] The virus inactivating agent according to claim 1, wherein the proanthocyanidin-based polyphenol is a polyphenol obtained from hops.

[5] The virus inactivating agent according to claim 4, wherein the hop is a hop cocoon.

6. The virus inactivating agent according to claim 1, containing 5 to 95% by weight of the proanthocyanin-based polyphenol.

[7] The virus inactivating agent according to [1], which is a proanthocyanin that does not permeate the membrane when treated with an ultrafiltration membrane having a molecular weight cut-off of 1,000 or more.

[8] A beverage product comprising the virus inactivating agent according to any one of claims 1 to 5 in a final concentration range of 1 to L0, 000 ppm.

[9] A food product comprising the virus inactivating agent according to any one of claims 1 to 5 in a final concentration range of 1 to L0, 000 ppm.

[10] A medicine comprising the virus inactivating agent according to any one of claims 1 to 5.

[II] A method of treatment, wherein the medicament according to claim 10 is administered as an oral or parenteral agent to a patient in a total daily dose of 2 mg to: LOOOmg.