WO2011005043A2 - Composition for the prevention and treatment of influenza virus infection and composition for suppressing neuraminidase activity comprising turmeric extract - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of influenza virus infection and a composition for suppressing neuraminidase activity comprising turmeric extract. Turmeric extract and turmeric fractions and curcuminoid compounds isolated therefrom exhibit an effect in suppressing neuraminidase activity, and simultaneously exhibit a virucidal and a cytopathy suppressing effect against various influenza viruses and can therefore be used advangageously in the prevention and treatment of influenza virus infection.

Description

Composition for the prevention and treatment of influenza virus infection comprising turmeric extract and composition for inhibition of neuraminidase activity

The present invention relates to a composition for preventing and treating influenza virus infection and a composition for inhibiting neuraminidase activity, including turmeric extract.

Influenza virus is a highly contagious virus that causes acute respiratory disease and is one of the viruses that causes severe respiratory symptoms in children, the elderly and patients with cardiopulmonary disease by causing a mass infection or pandemic all over the world (Hien, TT et al. N.) . Eng. J. Med., 350 , 1179, 2004). Influenza viruses belong to the orthomyxovirus taxonomically, and there are three types of A, B and C. In particular, the epidemic types are A and B. On the surface of these viruses, there are two types of surface antigens, glycoproteins, hemagglutinin (HA) and neuraminidase (NA), and there are eight segmented RNAs inside. Hemagglutinin is a trimer that consists of the head and the stem, the head of which is involved in most antigenic mutations and binds to the terminal sialic acid residues on the surface of the host cell to remove the virus. Attach and sequentially allow the virus to penetrate the host cell (Chandrasekaran, A. et al. Nature biotechnology 26, 107, 2008). Neuraminidase is a mushroom-shaped tetramer with a head and stem form, with active sites on the top surface of the head. Cleavage of alpha-ketosidic bonds, which link neuraminic acid residues, plays an important role in releasing viruses out of host cells and invading respiratory mucosal cells (a. Mark, VI Nature review 6, 967, 2007.b.Hermanerman, K. et al. Virology 214 , 294, 1995).

The surface antigens of the virus cause mutations in the same subtype, with new antigenic variants appearing each year. In particular, the avian influenza virus, which has been a problem until recently, is infected with various kinds of birds such as chickens, turkeys, ducks, and wild birds, and more than 80% of chickens die due to rapid spread. It is a virus disease that causes the greatest damage and threat to the poultry industry, and its ramifications are not limited to the poultry industry, and it has been reported to cause disease in humans due to infection with human body (Gubareva, LV et al. Lancet. 355 , 2000). To date, these influenza viruses have caused illnesses in humans.The three pandemic of the 20th century, the Spanish flu (H1N1), about 30 million people, the Asian flu (H2N2), about 1 million people, and the Hong Kong flu ( H3N2) has reported about one million deaths. In the 21st century, 385 people were killed and 243 died from 2003 to 2008. Recently, the H1N1 influenza was officially declared fender by the WHO, and as of June 29, 115 countries 70,893 people (including 311 deaths) occurred, and a sister who had volunteered in Mexico volunteered in Korea was confirmed as the first confirmed patient on May 2, 2009. The cumulative number of confirmed patients as of August 16 was 2,089 (2 deaths). Persons).

As a method for preventing and treating the infection of the virus, it can be considered that inhibition of adsorption into epithelial cells, inhibition of invasion into cells, inhibition of gene transcription and replication, inhibition of protein synthesis, inhibition of release from cells, etc. Each is a target of antiviral.

Conventionally, four substances, Amantadine, Rimantadine, Zanamivir and Oseltamivir, are approved by the US Food and Drug Administration (FDA) to treat diseases caused by influenza viruses. I receive it and am used. However, M2 inhibitors, Amatadine and Rimantadine, which act as antiviral agents by blocking the ion channel of M2 protein, which is a cell membrane protein essential for virus propagation, prevent the virus from uncoating. It has been reported to be effective and tolerate viruses for 40 years of use, with serious side effects in the nervous system and stomach (Bantia, S. et al. Antiviral Research 69 , 39, 2006). Since 1999, Zanamivir and Oseltamivir are inhibitors of neuraminidase, which play an important role in the propagation of viruses and have a low incidence of resistance and are reliably present in both influenza A and B influenza viruses. (Zhang, J. et al. Bioorg. Med. Chem. Lett. 16 , 3009, 2006).

However, zanamivir has a high antiviral effect but low bioavailability and rapid kidney discharge (Ryan, DM et al. Antimicrob. Agents Chemother., 39, 2583, 1995). Mir has the side effect of severe nausea.

Antivirals developed to date have severe side effects and require much attention for their application. In addition, the development of a vaccine has a problem that the effect is low if the virus of the virus and the type of the epidemic does not match, there is an increasing need for the development of a new influenza virus agent excellent in the inhibition of infection and excellent in stability.

Accordingly, the present inventors have confirmed that turmeric extracts and fractions or curcuminoid compounds isolated therefrom exhibited inhibitory activity against neuraminidase, virucidal inhibitory effect against virus and cytopathic inhibitory effect, and completed the present invention. .

It is an object of the present invention to provide a pharmaceutical composition for the prevention and treatment of influenza virus infection and a treatment method using the same.

Another object of the present invention is to provide a food composition for the prevention and improvement of influenza virus infection.

Still another object of the present invention is to provide a quasi-drug composition for the prevention and improvement of influenza virus infection.

It is still another object of the present invention to provide a composition for inhibiting neuraminidase activity and a method for inhibiting neuraminidase activity, which can be used to treat neuraminidase present in various organisms and to treat diseases related thereto.

Compositions comprising turmeric extract, fractions and curcuminoid compounds isolated therefrom exhibit an effect of inhibiting the activity of neuraminidase, and simultaneously show a virucidal and cytopathic effect against various influenza viruses Therefore, it can be usefully used for the prevention and treatment of influenza virus infection.

In one aspect of the present invention for achieving the above object, there is provided a composition for the prevention and treatment of influenza virus infection comprising turmeric extract or a fraction thereof as an active ingredient.

According to another aspect of the present invention, there is provided a composition for the prevention and treatment of influenza virus infection comprising a compound represented by the formula (1) as an active ingredient.

Formula 1

In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .

According to another aspect of the present invention, turmeric extract for the prevention and treatment of influenza virus infection, turmeric fraction or the use of the compound represented by the formula (1) or turmeric extract for the manufacture of a medicament for the prevention and treatment of influenza virus infection, The use of turmeric fraction or compound represented by the following formula (1) is provided.

According to another aspect of the present invention, a composition for the prevention and treatment of influenza virus infection comprising turmeric extract, turmeric fraction or a compound represented by the following formula (1) as an active ingredient to an individual having the onset or possibility of influenza virus infection Provided are methods for treating influenza virus infection comprising administering.

[Formula 1]

In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .

According to another aspect of the invention, there is provided a composition for inhibiting neuraminidase activity comprising turmeric extract, turmeric fraction or a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .

According to another aspect of the invention, there is provided a method of inhibiting the activity of neuraminidase comprising contacting the composition with a sample comprising neuraminidase.

Hereinafter, the present invention will be described in detail.

The present invention provides a composition for the prevention and treatment of influenza virus infection comprising turmeric extract or a fraction thereof as an active ingredient.

The composition of the present invention improves, prevents and treats influenza virus infections by inhibiting the influenza virus from spreading to other cells in the respiratory tract by inhibiting the activity of neuraminidase that is present on the surface of the influenza virus and plays an essential role in virus replication. Can be used for

As used herein, the term "prevention" means any action that inhibits or delays the influenza virus infection by administration of a composition.

As used herein, the term "treatment" means any action that improves or advantageously changes the symptoms caused by influenza virus infection by administration of the composition.

The influenza virus refers to an influenza virus strain that can cause disease in humans or animals, and may be influenza A virus, influenza B virus, influenza C virus, influenza A virus, influenza B virus, C virus. Type influenza viruses can be distinguished by their nuclear protein and matrix protein differences. The influenza A virus, influenza B virus, and influenza C virus also include their variants. Influenza virus subtypes are based on morphological differences between hemaglutinin (HA) and neuraminidase (NA). Currently, there are 6 types of HA (H1-H6) and 9 types of NA. (N1-N9). Preferably the influenza virus may be H1N1 influenza virus or H9N2 influenza virus. More preferably, the influenza virus may be rvH1N1 influenza A virus, H1N1 influenza virus (A / PR / 8/34) or H9N2 influenza virus (A / Chicken / Korea / MS96 / 96). In addition, the influenza virus may cause flu, cold, sore throat, bronchitis, and pneumonia, and in particular, may cause bird flu, swine flu, or chlorine flu.

Generally, turmeric refers to roots dried or dried by removing the bark of Curcuma longa Linne (Kep. 9th Amendment), and "lump root" is a root formed in the shape of a tube. Also called).

To mean a tuberous root portion of a plant belonging to turmeric (Turmeric) is keokyu equine (Curcuma sp.) In the present invention, the plant is on the turmeric belonging to keokyu equine (Curcuma sp.) (Curcuma wenyujin, Chen YH et C. Ling), autumn gold ( Curcuma longa Linne), spring gold ( Curcuma longa Salisb.), Purple gold ( Curcuma zedoaria ), gwangseok ( curcuma kwansiensis , SG Lee et CF Liang) Green grass ( Curcuma aeruginosa ), sprouts [ Curcuma phaeocaulis Val. (Ginger and Zingiberaceae) or Curcuma domestica , but are not necessarily limited thereto.

On the other hand, the root stem of Curcuma longa Linne is generally called Curcuma longa Rhizoma (9th Amendment), and "root stem" is an underground stem that grows as the roots of the plant grow into the ground. Also known as rhizome.

A plant belonging to Curcuma sp. Has a "root" and "root" area.

Specifically, as a result of measuring the content of the curcumin derivative according to the site of Curcuma longa Linne among the plants belonging to Curcuma sp. , Curcumin is contained as 17.0 g / kg ethanol extract in tuber. Demethoxycurcumin and bisdemethoxycurcumin are contained as 5.3 and 3.4 g / kg ethanol extracts, respectively. On the other hand, the compounds were not detected in the stem region, and the root stem region contained curcumin, demethoxycurcumin, and bisdemethoxycurcumin as 2.8, 0.4, and 6.8 g / kg ethanol extracts (Example 4).

Turmeric extract in the present invention can be extracted from a variety of organs of natural, hybrid, variety plants, preferably from tuber, plant tissue culture.

In the present invention, turmeric may be purchased and used commercially, or may be used collected or grown in nature.

According to another aspect of the present invention, there is provided a method for preparing a composition having neuraminidase activity comprising extracting turmeric with water, C ₁ to C ₄ alcohols or a mixed solvent thereof to obtain turmeric extract.

In addition, there is provided a method for producing a composition further comprising fractionating using hexane, ethyl acetate or water in the turmeric extract.

In addition, the method is provided for the preparation of a composition comprising separating and purifying the turmeric fraction.

The method of preparing the turmeric extract may use conventional extraction methods in the art, such as ultrasonic extraction, cold extraction, heat extraction, cold extraction, filtration and reflux extraction. The extraction method is not particularly limited, and may be extracted by room temperature or warming under conditions where the active ingredient is not destroyed or minimized. Also turmeric extract can be prepared using various solvents known in the art, for example, water, C_One~ C₄Alcohol, acetone, ethyl acetate, chloroform, or a mixed solvent thereof, but is not limited thereto. Preferably, the turmeric dried material obtained by pulverizing the roots of turmeric in which foreign substances have been removed by washing and drying the water, C_One~ C₄It may be an extract extracted with an alcohol or a mixed solvent of them, more preferably C_One~ C₄It may be an extract extracted with an alcohol, most preferably may be an extract extracted with methanol or ethanol. At this time, the extraction solvent is preferably 2 to 20 times the dry weight of turmeric. For example, cut the turmeric dry matter and put it in the extraction container._One~ C₄Lower alcohol or mixed solvents thereof, preferably methanol or ethanol, and allowed to stand at room temperature for a certain period of time, and then filtered to obtain an alcohol extract. At this time, the extraction is preferably left for 1 week at room temperature, after which it may be further subjected to methods such as concentration or lyophilization.

The extract of the present invention may include any one of an extract obtained by an extraction treatment, a diluent or concentrate of the extract, a dried product obtained by drying the extract, and these modifiers or purified products.

On the other hand, turmeric fraction in the present invention can be obtained by fractionation from the turmeric extract. In order to obtain fractions, various solvents known in the art can be used. For example, pentane, hexane, 2,2,4-trimethylpentane, dicaine, cyclohexane, carbon disulfide, carbon tetrachloride, chlorobutane, diisopropyl ether, chloroform, acetone, nitropropane, butanone, dichloroethane , Pyridine. Propanol, methanol, ethyl acetate, butanol and the like. More specifically, the turmeric fraction can be obtained by suspending the turmeric extract in water and fractionating with hexane and ethyl acetate, respectively, to obtain a hexane fraction, an ethyl acetate fraction and a water fraction.

Furthermore, the present invention provides a composition for the prevention and treatment of influenza virus infection comprising a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .

At this time, the compound of Formula 1 may be isolated from turmeric extract or turmeric fraction. More specifically, the compound represented by Chemical Formula 1 may be a compound represented by any one of the following Chemical Formulas 2 to 4.

Formula 2

Formula 3

Formula 4

In addition, the present invention a) extracting turmeric with water, C ₁ ~ C ₄ alcohol or a mixed solvent thereof to obtain turmeric extract; b) obtaining a hexane fraction, ethyl acetate fraction or water fraction from the extract; And c) provides a method for separating the compound represented by the formula (1) from turmeric comprising the step of separating and purifying the fraction.

More specifically, the compound represented by Chemical Formula 1 according to the present invention is a representative active ingredient present in turmeric, and the method for separating the compound may be performed as follows.

First, turmeric is extracted with water, C ₁ to C ₄ alcohol or a mixed solvent thereof to obtain turmeric extract (step 1). The turmeric can be used without limitation, such as cultivated or commercially available, it is used to clean and dry. The alcohol may be a lower alcohol such as methanol, ethanol, propanol, butanol, preferably methanol or ethanol.

In the step 1, the step of obtaining the turmeric extract is dried in the dry state of the turmeric and then made into a powder state to increase the extraction efficiency, put into an extraction container and add an appropriate amount of alcohol, and leave it at room temperature for 5 days And then filtered through a filter paper. This extraction process may be repeated several times, and then a method such as concentration or lyophilization may be additionally performed.

Subsequently, water is added to the turmeric extract obtained in step 1, suspended, and fractionated sequentially using hexane and ethyl acetate to obtain a turmeric fraction (step 2). At this time, a conventional fractional extraction method can be used, and preferably a fractional funnel can be used. The turmeric fraction may be obtained as a hexane fraction, an ethyl acetate fraction and a water fraction.

Thereafter, the turmeric fraction obtained in step 2 may be purified by separating and purifying the compound by silica gel chromatography (step 3). When performing silica gel chromatography for separating the compounds, it is preferable to use n -hexane, n -hexane ethyl acetate, chloroform and acetone mixed solvent and methanol as the mobile phase, and n -hexane and acetone mixed in the additional chromatography. Solvents may be used. In this case, the volume ratio of the n -hexane / ethyl acetate mixed solvent used is preferably 50: 1 to 1: 5, and preferably 150: 1 to 1: 4 in the case of the chloroform and acetone mixed solvent. The chromatography may be performed once or several times until a single compound is purified, and may be concentrated and recrystallized as necessary.

In addition, the compound represented by the formula (1) of the present invention can be used in the form of a pharmaceutically acceptable salt, the salt is an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids. Such pharmaceutically nontoxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and iodide. Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suverate , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, meth Oxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesul Nate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1- Sulfonates, naphthalene-2-sulfonates or mandelate.

The acid addition salts according to the present invention can be dissolved in conventional methods, for example, by dissolving the above formula (1) in an excess of aqueous acid solution and using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. It can be prepared by precipitation.

In addition, the compound represented by Formula 1 of the present invention can be used in the form of a pharmaceutically acceptable metal salt using a base. Alkali metal or alkaline earth metal salts are obtained, for example, by dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is pharmaceutically suitable to prepare sodium, potassium or calcium salt as the metal salt. Corresponding silver salts are also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

Meanwhile, the composition for preventing and treating influenza virus infection of the present invention may be a pharmaceutical composition.

When the composition of the present invention is a pharmaceutical composition, the composition may include a pharmaceutically acceptable carrier. The composition comprising a pharmaceutically acceptable carrier may be in various oral or parenteral formulations. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or lactose (at least one compound). lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, and syrups, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The pharmaceutical composition is any one selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, liquid solutions, emulsions, syrups, sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations and suppositories. It can have one formulation.

The composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level refers to the individual type and severity, age, sex, type of virus infected. , The activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of treatment, factors including the drug used concurrently, and other factors well known in the medical arts.

However, for the desired effect, turmeric extract or fractions thereof of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 100 mg / kg, and the compound of Formula 1 is 0.0001 to 100 mg / day. In kg, it is preferable to administer at 0.001 to 10 mg / kg. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. And single or multiple administrations. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, and can be easily determined by those skilled in the art.

The composition of the present invention can be used alone or in combination with methods for using surgery, hormonal therapy, drug therapy and biological response modifiers for the prevention and treatment of influenza virus infections.

In addition, the present invention is turmeric extract, turmeric fraction for the prevention and treatment of influenza virus infection or use of the compound represented by the formula (1) or turmeric extract, turmeric fraction or for the preparation of a medicament for the prevention and treatment of influenza virus infection It provides the use of the compound represented by the formula (1).

[Formula 1]

On the other hand, the present invention provides a method for the treatment of influenza virus infection disease comprising the step of administering the composition for the prevention and treatment of influenza virus infection in a pharmaceutically effective amount of the influenza virus infection to a subject having the onset or potential. . The influenza virus may be influenza A virus, influenza B virus, influenza C virus, preferably H1N1 influenza virus or H9N2 influenza virus. More preferably, the influenza virus may be rvH1N1 influenza A virus, H1N1 influenza virus (A / PR / 8/34) or H9N2 influenza virus (A / Chicken / Korea / MS96 / 96). In addition, the influenza virus infection disease may be flu, cold, sore throat, bronchitis, pneumonia, in particular bird flu, swine flu or goat flu.

As used herein, the term "individual" means all animals, including humans already infected with or capable of being infected with influenza virus, and effectively prevents the disease by administering to a subject a composition comprising an extract, fraction or compound of the present invention. And treatment. For example, the compositions of the present invention can treat humans infected with various influenza virus subtypes or variants of human influenza virus.

In addition, the compositions of the present invention can treat humans infected with various influenza virus subtypes or variants of avian influenza virus. It is also possible to treat chickens or pigs infected with various influenza virus subtypes or variants of avian influenza virus.

The composition of the present invention can be administered in parallel with a treatment for existing influenza virus infection disease. Examples include, but are not limited to, amatadine, rimantadine, zanamivir, or oseltamivir.

The route of administration of the composition may be administered via any general route as long as it can reach the desired tissue. The composition of the present invention may be administered as desired, but is not limited to intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, intranasal administration, pulmonary administration, rectal administration. The composition may also be administered by any device in which the active agent may migrate to the target cell.

In addition, the present invention provides a food composition for the prevention and improvement of influenza virus infection comprising turmeric extract, turmeric fraction or a compound represented by the following formula (1) isolated therefrom as an active ingredient.

[Formula 1]

In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .

At this time, the compound of Formula 1 may be isolated from turmeric extract or turmeric fraction. More specifically, the compound represented by Chemical Formula 1 may be a compound represented by any one of the following Chemical Formulas 2 to 4.

[Formula 2]

[Formula 3]

[Formula 4]

That is, the turmeric extract, turmeric fraction or the compound represented by Formula 1 isolated therefrom may be added to a food composition for the purpose of preventing or improving influenza virus infection.

The food composition may be a functional food according to the purpose of the present invention, wherein the 'functional food' is a food that emphasizes the bioregulatory function of the food, and acts and expresses a specific purpose using physical, biochemical and biotechnological methods. It is food added with added value. The components of the functional foods are designed and processed to fully exercise the body control functions related to the biological defense and regulation of the body rhythm, prevention and recovery of diseases to the living body.

When using turmeric extract, fraction or compound represented by Formula 1 separated therefrom as a food additive, the extract, fraction or compound may be added as it is or used with other food or food ingredients, according to a conventional method. Can be used as appropriate. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). In general, the turmeric extract, turmeric fraction or the compound represented by Formula 1 separated therefrom in the preparation of food or beverage is in an amount of 0.01 to 10% by weight, preferably 0.05 to 1% by weight in the raw material composition. Is added. However, in the case of prolonged ingestion for health and hygiene purposes or for health control purposes, the amount may be used below the above range.

There is no particular limitation on the kind of food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, And a carbonation agent used for the carbonated beverage. The proportion of such additives is not critical but is usually selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention. In addition, the composition of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. The proportion of such pulp is not critical, but is generally selected in the range of 0.01 to 10 parts by weight per 100 parts by weight of the composition of the present invention. These components can be used independently or in combination.

In addition, the present invention provides a quasi-drug composition for the prevention and improvement of influenza virus infection comprising turmeric extract, turmeric fraction or a compound represented by the following formula (1) isolated therefrom as an active ingredient.

[Formula 1]

In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .

At this time, the compound of Formula 1 may be isolated from turmeric extract or turmeric fraction. More specifically, the compound represented by Chemical Formula 1 may be a compound represented by any one of the following Chemical Formulas 2 to 4.

[Formula 2]

[Formula 3]

[Formula 4]

That is, the composition of the present invention may be added to the quasi-drug composition for the purpose of preventing or improving influenza virus infection. When the turmeric extract, fraction or compound represented by Formula 1 separated therefrom is used as an quasi-drug additive, the extract, fraction or compound may be added as it is, or may be used together with other quasi-drug or quasi-drug components, according to a conventional method. Can be used as appropriate. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment).

Preferably, the quasi-drug composition may be used in the manufacture of natural disinfectants, feed additives, antiseptic cleaners, shower foams, gagreens, wet wipes, detergent soaps, hand washes, humidifier fillers, masks, ointments or filter fillers.

In addition, the present invention provides a composition for inhibiting neuraminidase activity comprising turmeric extract, turmeric fraction or a compound represented by the following formula (1) as an active ingredient.

[Formula 1]

In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .

At this time, the neuraminidase may be derived from influenza A virus, influenza B virus, influenza C virus, Clostridium perfringens or human, preferably H1N1 influenza virus or H9N2 influenza. It may be from a virus. More preferably, the influenza virus may be derived from rvH1N1 influenza A virus, H1N1 influenza virus (A / PR / 8/34) or H9N2 influenza virus (A / Chicken / Korea / MS96 / 96).

Neuraminidase (also known as sialidase, acylneuraminil hydrolase) is an enzyme present in animals and some microorganisms, and many microorganisms containing neuraminideniz are found in humans, poultry, horses, pigs, and It causes sickness in seals and other animals. Therefore, the composition of the present invention that inhibits neuraminidase activity can be usefully used for the prevention and treatment of many diseases related to the activity of neuraminideniz.

The present invention also provides a method of inhibiting the activity of neuraminidase comprising contacting the composition with a sample comprising neuraminidase. At this time, the contact is made in a conventional manner, the sample containing neuraminidase is a living organism, tissue or cell culture, biological material samples (blood, serum, urine, cerebrospinal fluid, tears, sputum, saliva, tissue samples, etc.) And the like, and may contain organisms that produce neuraminidase, usually pathogenic organisms such as viruses. Such samples may be contained in any medium, including water, organic solvent / water mixtures, and the like.

In addition, the activity of neuraminidase after administration of the composition of the present invention can be observed by any method including a direct or indirect method of diagnosing neuraminidase activity. Either quantitative, qualitative or semi-quantitative methods of diagnosing neuraminidase activity are available, and any other method may be applied, such as observing the physiological properties of living organisms.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the following examples are merely to illustrate the present invention is not limited to the contents of the present invention.

Example 1 Preparation of Turmeric Extract

The turmeric used in the present embodiment is generally available in Chinese medicine or on the market and is dried in the form of dried Root of Curcuma longa Linne, and then dried to obtain the extract of the present invention efficiently in powder form. It was used by grinding. 7.5 L of 100% ethanol (EtOH) was added to 1.6 kg of turmeric, and the mixture was left to stand at room temperature for 5 days, filtered through a filter paper, and concentrated to obtain turmeric ethanol extract (170 g).

Example 2: Isolation and Purification of the Curd fraction and Curcuminoid Compounds from the Curd Extract

170 g of ethanol extract of turmeric obtained in Example 1 was added and suspended in 1 L of water. This was put into a separatory funnel, and fractional extraction using n -hexane and ethyl acetate in order to obtain n -hexane soluble extract (23 g), ethyl acetate soluble extract (85 g) and water soluble extract (34 g).

85 g of the ethyl acetate soluble extract obtained above was subjected to silica gel column chromatography [silica gel 500 g, 70 to 230 mesh] using chloroform, methanol, and a mixed solvent thereof (80: 1 to 1: 1) as a mobile phase. 15 fractions (Fr.-1-15) were separated. The sixth fraction (Fr.-6, 16 g) was purified by silica gel column chromatography using a mixed solvent of n -hexane: ethyl acetate (20: 1 ~ 1: 1 (v / v)) as a mobile phase (30 g, 230 to 400 mesh) was carried out to obtain five fractions (Fr.-6-1-5).

Among the fractions obtained by performing silica gel column chromatography on Fr.-6-2-3 fractions (11 g) with chloroform, methanol and their mixed solvents (80: 1 to 4: 1) as mobile phases. Development by preparative TLC method [( n -hexane: ethylacetate = 4: 1 (v / v)] gave pure compound 1 (8 g), and the eighth fraction (Fr.-8, 14 g) is silica gel using a mixed solvent of n -hexane: ethyl acetate (20: 1 to 1: 1 (v / v)) and chloroform: methanol (80: 1 to 20: 1 (v / v)) as a mobile phase. Column chromatography (30 g, 230-400 mesh) was repeatedly performed to yield compound 2 (0.4 g) and 3 (0.2 g).

Example 3: Structural Analysis of Curcuminoid Compounds

The molecular weight and molecular formula of the curcuminoid compound obtained in Example 2 were determined using a VG high resolution GC / MS spectrometer, Election Ionization MS, Autospec-Ultima. In addition, molecular structure was determined using ¹ H-NMR, ¹³ C-NMR and 2D NMR spectroscopy data through nuclear magnetic resonance (NMR) analysis (Bruker AM 500).

As a result of comparing the results of the instrument analysis with that of the published literature, curcumin, demethoxycurcumin and bisdemethoxycurcumin represented by the following Chemical Formulas 2 to 4 were identified ( Food Chem . 265-272, 2009; J. Nat Prod . 1227-1231, 2002; J. Nat. Prod . 1531-1534, 1998; J. Agric.Food Chem . 3668-3672, 2002). The specific analysis results are as follows.

Compound 1: Curcumin

[Formula 2]

1) Physical property: bright orange powder (m.p. 183 ℃)

2) Molecular weight: 368.3

3) Molecular formula: C ₂₁ H ₂₀ O ₆

4) ¹ H-NMR (acetone- d ₆ , 500 MHz) δ 7.62 (2H, d, J = 15.80 Hz, H-4, H-4 '), 7.35 (2H, d, J = 1.91 Hz, H- 6, H-6 '), 6.83 (2H, H-3, H-5), 7.20 (2H, dd, J = 8.3, 1.9 Hz, H-10, H-10'), 6.90 (2H, d, J = 8.15 Hz, H-9, H-6 '), 5.99 (1H, s, H-1). ¹³ C-NMR (acetone- d ₆ , 125 MHz) δ 56.72, 102.01, 111.95, 116.64, 122.72, 124.25, 128.58, 141.81, 149.20, 150.44, 184.94.

Compound 2: Demethoxycurcumin

[Formula 3]

1) Physical property: Orange powder (m.p. 220 ℃)

2) Molecular Weight: 338

3) Molecular formula: C ₂₀ H ₁₈ O ₅

4) ¹ H-NMR (acetone- d ₆ , 500 MHz) δ 7.62-7.55 (4H), 7.34 (1H), 7.18 (1H), 6.89 (3H), 6.70 (2H), 5.97 (1H). ¹³ C-NMR (acetone- d ₆ , 125 MHz) δ 56.38, 101.79, 111.53, 116.30, 116.89, 122.12, 122.35, 123.98, 127.77, 128.24, 131.06, 141.13, 141.48, 148.87, 150.13, 160.64, 184.66.

Compound 3: Bisdemethoxycurcumin

[Formula 4]

1) Physical property: Orange powder (m.p. 224 ℃)

2) Molecular Weight: 308

3) Molecular formula: C ₁₉ H ₁₆ O ₄

4) ¹ H-NMR (acetone- d ₆ , 500 MHz) δ 7.62-7.56 (6H), 6.91-6.87 (4H), 6.68-6.65 (2H), 5.98 (1H). ¹³ C-NMR (acetone- d ₆ , 125 MHz) δ 101.82, 116.87, 122.11, 127.79, 131.06, 141.12, 160.58, 184.62.

Example 4: HPLC analysis of turmeric alcohol extract and fractions

HPLC chromatograms of turmeric extracts, fractions and curcuminoid compounds obtained from Examples 1 and 2 were performed. The HPLC analysis used in this example used our pump system with an Agilent 1200 Series HPLC instrument. The detector used in this example was tested using the company's variable wavelength detector (VWD) at the best analytical detection wavelength of 260 nm, and the solvent flow rate was 1.0 ml / min. The sample injection amount was 10 μl, and was prepared at a concentration of 10 mg / ml each. HPLC columns were analyzed using ZORBAX-SB-18 (5 μm, 150 mm × 4.6 mm). Analysis of the mobile phase was performed with 20% acetonitrile, 0 min; under polarization conditions of water (containing 0.1% TFA) and acetonitrile (containing 0.1% TFA); 25% acetonitrile, 10 min; 35% acetonitrile, 20 min; 50% acetonitrile, 30 min; 60% acetonitrile, 40 min; 70% acetonitrile, 50 min; 100% acetonitrile, 60 min.

Curcumin, demethoxycurcumin and bisdemethoxycurcumin concentrations of 15.625, 31.25, 62.5, 125, 250, 500 It was prepared at 1000 ㎍ / ml and analyzed under the same conditions to obtain a calibration line.

Curcumin, demethoxycurcumin and bisdemethoxycurcumin contents of the turmeric extracts and fractions were measured based on the calibration lines.Curcumin 106 g / kg (extract) and demethoxy curcumin (45.4 g / kg) , 64.6 g / kg of bisdemethoxycurcumin (extract), 114.3 g / kg of curcumin (fraction) in the ethyl acetate fraction, 45.8 g / kg of demethoxycurcumin (fraction), 66.4 g / kg of fraction (bis) and hexane The fractions contained curcumin 21.3 g / kg (fraction), demethoxycurcumin 7.3 g / kg (fraction) and bisdemethoxycurcumin 31.2 g / kg (fraction).

Quantitative results are shown in Table 1.

Table 1

In addition, the curcumin, demethoxy curcumin and bisdemethoxy curcumin content of the alcohol extract for each region of Curcuma longa Linne based on the assay line was measured, Curcumin 17.0 g / kg (extract), deme 5.3 g / kg of oxycurcumin (extract) and 3.4 g / kg (extract) of bisdemethoxycurcumin were detected, and curcumin, demethoxycurcumin and bisdemethoxycurcumin were not detected at the stem site. In the root stem region, curcumin 2.8 g / kg (fraction), demethoxy curcumin 0.4 g / kg (fraction), and bisdemethoxy curcumin 6.8 g / kg (fraction) were shown.

Quantitative results are shown in Table 2.

TABLE 2

Experimental Example 1 Determination of Neuraminidase A / Bervig-Mission / 1/18 (rvH1N1) Inhibitory Activity of Turmeric Extract, Fraction and Curcuminoid Compounds

In order to measure the inhibitory activity against the neuraminidase of turmeric extracts, turmeric fractions and curcuminoid compounds isolated from Examples 1 and 2 of the present invention, the virus isolated from Spanish flu in 1918 (A / Bervig_Mission / 1/18) of neuraminidase (R & D SYSTEM, 4858-NM) of the rvH1N1 influenza A virus, a recombinant neuraminidase. The substrate is 2 '-(4-trimethylumbeliferyl) -α-D- N -acetyl-neuraminic acid sodium salt [2'-(4-trimethylumbelliferyl) -α-D- N -acetyl-neuraminic acid sodium salt] Was purchased from Sigma.

Turmeric extracts of Examples 1 and 2 and fractions thereof were dissolved in methanol and added to each 20 μL, and as a substrate, 2 ′-(4-trimethylumbeliferyl) -α-D- N -acetyl-neuraminic acid sodium salt ( 50 μL of the final concentration (200 μM) was added and mixed with 80 μL of Tris buffer solution (pH 7.5) added with 5 mM CaCl ₂ and 200 mM NaCl, and the enzyme source neuraminidase (enzyme final concentration, 0.05 ng / μL). The inhibitory activity of neuraminidase was determined by adding 50 μL to react at room temperature for 25 minutes at 25 ° C. and measuring the absorption at 365 nm and the emission at 445 nm with a fluorescence spectrometer.

The measurement results are shown in Table 3 below.

TABLE 3

As shown in Table 3, as a result of measuring the inhibitory activity against the neuraminidase of the turmeric extract and fractions, curcuminoid-based compounds according to the present invention, the turmeric ethanol extract is 3.1 for the neuraminidase of influenza virus source The IC ₅₀ value of μg / mL was shown, the turmeric hexane fraction showed 262.0 μg / mL, the turmeric ethyl acetate fraction 0.9 μg / mL, and the turmeric water fraction showed an IC ₅₀ value of 61.5 μg / mL. In addition, curcuminoid compounds curcumin and demethoxy curcumin showed an IC ₅₀ value of 3.0 μM, bisdemethoxy curcumin 6.0 μM. Therefore, it was confirmed that the turmeric extract, fractions thereof, and curcuminoid compounds according to the present invention have excellent neuraminidase inhibitory activity through the above results.

Experimental Example 2 Determination of the Inhibitory Effect of Curcuminoids, Ethyl Acetate Fraction, and Ethanol Extracts Isolated from Turmeric Extracts against Influenza Virus

First, the killer virus against influenza viruses H1N1 (A / PR / 8/34) and H9N2 (A / Chicken / Korea / MS96 / 96)] of curcuminoid compound, ethyl acetate fraction or ethanol extract isolated from turmeric extract In order to measure the effect, the following in vitro experiments were performed using MDCK (Madin-Darby canine kidney, ATCC: CCL-34), which is a dog kidney cell line.

First, MDCK cells were put in a 96 well microplate to 1 X 10 ⁵ / well for each well, and cultured in medium EMEM (penicillin 100 units, streptomycin 100 ㎍, 10% FBS). When MDCK cells became monolayers, the cells were washed twice with EMEM medium containing only antibiotics. H1N1 and H9N2 strains were diluted to 100 TCID ₅₀ and placed in EP tubes. Curcuminoid compounds, ethyl acetate fractions or ethanol extracts isolated from turmeric extract diluted with DMSO (dimethylsulfoxide) were added to each tube by concentration and reacted at 4 ° C. for 1 hour. After 1 hour, each well was inoculated with 3 wells per concentration to the MDCK cells washed in advance, and incubated at 35 ° C. for 1 hour (hereinafter referred to as a sample treatment group). On the other hand, after infection with the non-infected + non-administered control group (cell group not infected with Control, H1N1 or H9N2 strain, and without curcuminoid compound) and the infection + non-administered control group (Virus control, H1N1 or H9N2 strain), the curcumin Cell groups not administered) were inoculated into MDCK cells under the same conditions and incubated at 35 ° C. for 1 hour. After 1 hour, all of the plate medium was removed and washed once with PBS. After 100 ml of EMEM medium with antibiotics and 10 ㎍ / mL trypsin was added to each well, the cells were incubated at 35 ° C for 48-72 hours. Infected + non-administered controls were incubated for 48-72 hours until complete cytopathic effect (CPE). Cell state was observed daily by inverted microscope. To determine cell viability after 48-72 hours of incubation, 10 ml of each cell counting kit-8 (Dojin, Kumanoto, Japan, tetrazolium salt WST-8) was added to each well, followed by reaction at 35 ° C for 2 hours, and then at 450 nm. Absorbance was measured. At this time, the virucidal effect (lower percent (%)) exhibited by the curcuminoid compound, ethyl acetate fraction or ethanol extract isolated from the non-infected + non-administered control and the infection + non-administered control and turmeric extract of the present invention After the comparison by 1, the results are shown in Table 4 below.

Equation 1

In the above formula, the O.D value means the absorbance value measured at 450 nm.

On the other hand, cell degeneration of influenza viruses H1N1 (A / PR / 8/34) and H9N2 (A / Chicken / Korea / MS96 / 96)] of curcuminoid compound, ethyl acetate fraction and ethanol extract isolated from turmeric extract In order to determine the inhibitory effect, the influenza virus (H1N1 or H9N2 strain) was inoculated in MDCK cells washed twice with EMEM medium containing only antibiotics and incubated at 35 ° C for 1 hour. After 1 hour, all the inoculated virus solution was removed, and curcuminoid compounds, ethyl acetate fractions and ethanol extracts isolated from turmeric extract were placed in MDCK cells inoculated with the virus. Thereafter, the cell denaturation inhibitory effect (inferiority (%)) represented by the curcuminoid compound, the ethyl acetate fraction, and the ethanol extract isolated from the turmeric extract of the present invention in the same manner as described above, The results are shown in Table 4 below.

Table 4

a, mixed with curcuminoid compound, ethyl acetate fraction or ethanol extract and virus for 1 hour at 4 ℃, 1 hour after infection with MDCK cells, washed once with PBS and EPS medium with 10 mg / ml of trypsin added Incubate for 48-72 hours at 35 ° C.

b, 1 hour after the virus was infected with MDCK cells, the medium containing the virus was removed and the medium containing the curcuminoid compound, ethyl acetate fraction or ethanol extract was incubated for 48-72 hours.

c, a value of CC ₅₀ / EC ₅₀ as the selection index.

As shown in Table 4, the curcuminoid compounds showed excellent virucidal effects with selective indexes (SIs) of 13.3, 23.1, and 7.1 showing the virucidal effect in H1N1 strains. In addition, H9N2 also exhibited virucidal effects on various virus strains, showing the choices below 5.1, 5.3 and 1.0. The turmeric ethyl acetate fraction showed a very good virucidal effect with a selection index of 9.1 in H1N1 strain and 6.4 in turmeric ethanol extract. In addition, as a result of observing the cell morphology on the inverted microscope, the virus-inoculated group (H1N1, H9N2) showed almost 90-100% cell denaturation effect due to almost destruction of MDCK cells, but the virus, curcuminoid compound, and ethyl acetate The sample treated group treated with the fraction or ethanol extract was confirmed to be similar to the non-infected + non-administered control group treated with nothing in the MDCK cells.

In addition, curcuminoid compounds showed inhibitory effects on H1N1 and H9N2 cell degeneration. In particular, H9N2 strains showed very good cytopathic inhibition effect with selection index 8.6, 3.9 and 1.1, and H1N1 strains showed cytopathic inhibition effect with selection index 2.3, 1.6 and 1.4 or higher. In addition, as a result of observing the cell morphology on the inverted microscope, the virus-inoculated group (H1N1, H9N2) showed almost 90-100% cell denaturation effect due to almost destruction of MDCK cells, but the virus, curcuminoid compound, and ethyl acetate The group treated with the fraction or ethanol extract was confirmed to be similar to the non-infected + non-administered control group treated with nothing in the MDCK cells.

Therefore, the composition of the present invention exhibits a virucidal effect by directly acting on the virus before the virus is infected with the cell, and also exhibits an excellent inhibitory effect on cellular denaturation by preventing the virus from being released into the cell after replication. Therefore, it can be usefully used for the prevention and post-infection treatment of influenza virus infection.

Experimental Example 3: Acute Toxicity Test of Compounds of the Invention

In order to determine the acute toxicity for the compounds of the present invention, the following experiment was performed.

SPF (specific pathogens free) 6-week-old C57BL / 6J mice were divided into four groups (three males and three females / experimental group) of 12 males and females, temperature 22 ± 3 ° C, humidity 55 ± 10%, illumination 12L / The animals were bred in 12D animal rooms. Mice were allowed to acclimate for about a week before being used in the experiment. Feed for experimental animals (for mice and rats, CheilJedang Co., Seoul, South Korea) and drinking water were sterilized and supplied freely.

Compounds represented by Formulas 2 to 4, prepared in Example 2, were prepared at a concentration of 50 mg / mL in 0.5% tween 80, respectively, and then 0.04 mL (100 mg / kg) per 20 g of mouse body weight. 0.2 mL (500 mg / kg) and 0.4 mL (1,000 mg / kg) were administered orally. Samples were administered orally once and observed for side effects or lethality for 7 days after administration. That is, on the day of administration, changes in general symptoms and the presence or absence of dead animals were observed at least once in the morning, at least once every afternoon from 1 hour, 4 hours, 8 hours, 12 hours, and the next day after administration.

As a result of the acute toxicity test as described above, all mice treated with the sample did not show any significant clinical symptoms, and there were no dead mice. Also, no change in toxicity was observed in weight change, blood test, blood biochemical test, autopsy findings, etc. Did.

Therefore, the compounds of the present invention did not show a toxic change up to 1,000 mg / kg in all mice, it was confirmed that oral administration minimum dose (LD ₅₀ ) is a safe substance more than 1,000 mg / kg.

Hereinafter, the preparation of pharmaceutical preparations or health foods containing turmeric extract, turmeric fraction, curcuminoid compounds isolated therefrom or pharmaceutically acceptable salts thereof will be described.

Preparation Example 1 Preparation of Pharmaceutical Formulation

1-1. Manufacture of powder

Turmeric extract, fractions, compounds isolated therefrom or salts thereof 2 g

1 g lactose

After mixing the above components, the airtight cloth was filled to prepare a powder.

1-2. Manufacture of tablets

Turmeric extract, fractions, compounds isolated therefrom or salts thereof 100 mg

100 mg corn starch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

1-3. Preparation of Capsule

Turmeric extract, fractions, compounds isolated therefrom or salts thereof 100 mg

100 mg corn starch

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

1-4. Preparation of Injection

Turmeric extract, fractions, compounds isolated therefrom or 10 μg / ml thereof

Dilute hydrochloric acid BP until pH 3.5

Injectable sodium chloride BP up to 1 ml

Turmeric extract, fractions, compounds isolated therefrom or salts thereof are dissolved in an appropriate volume of injectable sodium chloride BP, and the pH of the resulting solution is adjusted to pH 3.5 with dilute hydrochloric acid BP and injectable sodium chloride BP The volume was adjusted and mixed well. The solution was filled into a 5 ml Type I ampoule made of clear glass, encapsulated under an upper grid of air by dissolving the glass, and sterilized by autoclaving at 120 ° C. for at least 15 minutes to prepare an injection solution.

Preparation Example 2 Preparation of Health Food

2-1. Preparation of Cooking Seasonings

Turmeric extract, fractions, compounds isolated therefrom or salts thereof prepared from 0.2 to 10% by weight for health promotion cooking seasoning.

2-2. Preparation of Tomato Ketchup and Sauce

Health promotion tomato ketchup or sauce was prepared by adding 0.2-1.0 wt% turmeric extract, fraction, compound isolated therefrom or salt thereof to tomato ketchup or sauce.

2-3. Manufacture of Flour Food

Turmeric extract, fractions, 0.1 to 5.0% by weight of the compounds or salts thereof were added to the flour, and bread, cake, cookies, crackers and noodles were prepared using the mixture to prepare health promoting foods.

2-4. Preparation of soups and gravy

Turmeric extract, fractions, 0.1 to 1.0% by weight of the compounds or salts thereof were added to soups and broth to prepare health products, health soups and broths.

2-5. Preparation of Ground Beef

Health promotion ground beef was prepared by adding turmeric extract, fractions and 10% by weight of a compound or salt thereof separated to the ground beef.

2-6. Manufacture of dairy products

Turmeric extract, fractions, compounds isolated therefrom or salts thereof from 0.1 to 1.0% by weight were added to the milk, using the milk to prepare a variety of dairy products such as butter and ice cream.

2-7. Manufacture of wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh.

Black beans, black sesame seeds, and perilla were also steamed and dried in a known manner, and then roasted to prepare a powder having a particle size of 60 mesh.

The turmeric extract, fractions and the compounds or salts thereof separated from the turmeric extract were concentrated under reduced pressure in a vacuum concentrator, dried by spraying and drying with a hot air dryer, and then pulverized with a particle size of 60 mesh to obtain a dry powder.

Grains prepared above; Seed species; And turmeric extract, fractions, and the dry powder of a compound or salt thereof separated from the mixture in the following ratio.

75% by weight of grains (brown rice 35%, barley 15%, barley 25%)

23% by weight of seeds (7% by weight perilla, 9% by weight black beans, 7% by weight black sesame seeds)

1% by weight of turmeric extract, fractions, compounds isolated therefrom or salts thereof

Ganoderma lucidum 0.5% by weight

Yogurt 0.5% by weight

2-8. Preparation of Carbonated Drinks

5 to 10% by weight of sugar, 0.05 to 0.3% by weight of citric acid, 0.005 to 0.02% by weight of caramel, 0.1 to 1% by weight of vitamin C are mixed, and purified water is mixed to make the total composition 100% by weight to make syrup. After sterilizing the syrup at 85 to 98 ° C. for 20 to 180 seconds, mixing the cooling water with a volume ratio of 1: 4 to prepare a beverage composition, and injecting carbon dioxide into 0.5 to 0.82% by volume of the beverage composition, turmeric extract , Carbonated beverages containing fractions, compounds isolated therefrom or salts thereof were prepared.

2-9. Manufacture of health drinks

Substances such as liquid fructose (0.5 wt%), oligosaccharide (2 wt%), sugar (2 wt%), salt (0.5 wt%), water (94 wt%) and turmeric extracts, fractions, compounds isolated therefrom or Its salt (1% by weight) was homogeneously blended and sterilized immediately and then packaged in a small packaging container such as a glass bottle or a plastic bottle to prepare a health beverage.

2-10. Preparation of Vegetable Juice

Turmeric extract, fractions, 0.5 g of the compounds or salts thereof were added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

2-11. Preparation of Fruit Juice

0.1 g of turmeric extract, fractions, compounds isolated therefrom or salts thereof were added to 1,000 ml of apple or grape juice to prepare fruit juices for health promotion.

Claims (32)

1.  A composition for the prevention and treatment of influenza virus infection comprising turmeric extract or a fraction thereof as an active ingredient.
2.  The method of claim 1, wherein the turmeric is warm turmeric (Curcuma                                  wenyujin, Y. H. Chen et C. Ling), autumn gold (yellow gold) (Curcuma                                  longa Linne), spring grass (Curcuma                                  longa Salisb.), Purple turquoise (Curcuma zedoaria), Guangxi Sunrise (Graduate) (Curcuma kwansiensis, S. G. Lee et C. F. Liang), AchulCurcuma aeruginosa), Buds [Curcuma phaeocaulis Val. (Ginger and Zingiberaceae)] or Curcuma dometica (Curcuma domesticaA composition for preventing and treating influenza virus infection, characterized in that).
3.  According to claim 1, wherein the turmeric is a composition for the prevention and treatment of influenza virus infection, characterized in that the tuber roots.
4. According to claim 1, wherein the extract is a composition for the prevention and treatment of influenza virus infection, characterized in that the extraction using water, C ₁ ~ C ₄ alcohol or a mixed solvent thereof.
5.  The composition for preventing and treating influenza virus infection according to claim 1, wherein the fraction is a hexane fraction, an ethyl acetate fraction or a water fraction.
6.  The method of claim 1, wherein the influenza virus is influenza A virus, influenza B virus or influenza C virus, characterized in that the composition for the prevention and treatment of influenza virus infection.
7.  The composition of claim 1, wherein the influenza virus is H1N1 influenza virus or H9N2 influenza virus.
8.  The influenza virus infection of claim 1, wherein the influenza virus is rvH1N1 influenza A virus, H1N1 influenza virus (A / PR / 8/34) or H9N2 influenza virus (A / Chicken / Korea / MS96 / 96). Composition for the prevention and treatment of.
9. According to claim 1, wherein the disease caused by influenza virus infection is flu, cold, sore throat, bronchitis, pneumonia, bird flu, swine flu or chlorine flu composition for the prevention and treatment of influenza virus infection.
10. The method of claim 1, wherein the composition is a pharmaceutical composition, a food composition or a quasi-drug composition for the prevention and treatment of influenza virus infection, characterized in that.
11. The pharmaceutical composition of claim 10, wherein the pharmaceutical composition is a tablet, pill, powder, granule, capsule, suspension, liquid solution, emulsion, syrup, sterilized aqueous solution, non-aqueous solvent, suspension, emulsion, lyophilized agent and suppository. A composition for the prevention and treatment of influenza virus infection, characterized in that it has any one formulation selected from the group consisting of.
12. The method of claim 10, wherein the food composition is from the group consisting of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes Composition for the prevention and treatment of influenza virus infection, characterized in that any one selected.
13. The quasi-drug composition according to claim 10, wherein the quasi-drug composition is selected from the group consisting of natural disinfectants, feed additives, disinfectant cleaners, shower foams, gagrins, wet wipes, detergent soaps, hand washes, humidifier fillers, masks, ointments, and filter fillers. Composition for the prevention and treatment of influenza virus infection, characterized in that used in the manufacture.
14. A composition for the prevention and treatment of influenza virus infection comprising the compound represented by the formula (1) as an active ingredient.

    [Formula 1]

    

    In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .
15. 15. The composition for preventing and treating influenza virus infection of claim 14, wherein the compound represented by Chemical Formula 1 is a compound represented by any one of the following Chemical Formulas 2 to 4.

    [Formula 2]

    

    [Formula 3]

    

    [Formula 4]

    
16. The composition for preventing and treating influenza virus infection according to claim 14, wherein the influenza virus is an influenza A virus, an influenza B virus, or an influenza C virus.
17. 15. The composition for preventing and treating influenza virus infection according to claim 14, wherein the influenza virus is H1N1 influenza virus or H9N2 influenza virus.
18. 15. The composition for preventing and treating influenza virus infection according to claim 14, wherein the disease caused by influenza virus infection is flu, cold, sore throat, bronchitis, pneumonia, bird flu, swine flu or goat flu.
19. The method of claim 14, wherein the composition is a pharmaceutical composition, a food composition or a quasi-drug composition for the prevention and treatment of influenza virus infection, characterized in that.
20. The pharmaceutical composition of claim 19, wherein the pharmaceutical composition is a tablet, pill, powder, granule, capsule, suspension, liquid solution, emulsion, syrup, sterilized aqueous solution, non-aqueous solvent, suspension, emulsion, lyophilized agent and suppository. A composition for the prevention and treatment of influenza virus infection, characterized in that it has any one formulation selected from the group consisting of.
21. The method of claim 19, wherein the food composition is from the group consisting of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes Composition for the prevention and treatment of influenza virus infection, characterized in that any one selected.
22. The quasi-drug composition according to claim 19, wherein the quasi-drug composition is selected from the group consisting of natural disinfectants, feed additives, disinfectant cleaners, shower foams, gagrins, wet wipes, detergent soaps, hand washes, humidifier fillers, masks, ointments, and filter fillers. Composition for the prevention and treatment of influenza virus infection, characterized in that used in the manufacture.
23. Turmeric extract, turmeric fraction or a composition for inhibiting neuraminidase activity comprising a compound represented by the formula (1) as an active ingredient.

    [Formula 1]

    

    In the above formula, each R independently represents hydrogen, a hydroxy group, or an alkoxy group of C ₁ to C ₁₀ .
24. The method according to claim 23, wherein the neuraminidase is derived from influenza A virus, influenza B virus, influenza C virus, Clostridium perfringens , or human Composition.
25. The composition for inhibiting neuraminidase activity according to claim 23, wherein the neuraminidase is derived from H1N1 influenza virus or H9N2 influenza virus.
26. Influenza virus infection comprising the step of administering a composition for the prevention and treatment of influenza virus infection comprising turmeric extract, turmeric fraction or a compound represented by the following formula (1) as an active ingredient: Method of treatment.

    [Formula 1]

    
27. A method for inhibiting neuraminidase activity, comprising contacting a composition for preventing and treating influenza virus infection comprising turmeric extract, turmeric fraction, or a compound represented by the following Formula 1 as an active ingredient with a sample comprising neuraminidase.

    [Formula 1]

    
28. a) extracting turmeric with water, C ₁ -C ₄ alcohol or a mixed solvent thereof to obtain turmeric extract; b) obtaining a hexane fraction, ethyl acetate fraction or water fraction from the extract; And c) separating the compound represented by the following Chemical Formula 1 from the turmeric comprising the step of separating and purifying the fraction.

    [Formula 1]

    
29. 29. The method of any one of claims 26 to 28, wherein the compound represented by Chemical Formula 1 is a compound represented by any one of the following Chemical Formulas 2 to 4.

    [Formula 2]

    

    [Formula 3]

    

    [Formula 4]

    
30. A method of preparing the composition of claim 1 having a neuraminidase activity comprising extracting turmeric with water, C ₁ to C ₄ alcohols or a mixed solvent thereof to obtain turmeric extract.
31. 31. The method of claim 30, further comprising fractionating the turmeric extract with hexane, ethyl acetate or water.
32. 32. The method of claim 31, comprising separating and purifying the turmeric fraction.