KR101846425B1 - Composition for preventing or treating of influenza virus infection comprising the extract of Glaziella splendens and Compound isolated from the extract - Google Patents

Abstract

The present invention relates to a composition for preventing or treating influenza virus infection comprising peanut mushroom extract and an active substance isolated therefrom as an active ingredient. The peanut mushroom extract of the present invention and the compounds of the general formulas (1) to (3) The present invention is useful as a pharmaceutical composition and food for preventing or treating an influenza virus infection because it has an excellent effect of preventing or treating influenza virus infection by inhibiting the activity of neuraminidase enzyme of influenza virus.

Description

[0001] The present invention relates to a composition for prevention or treatment of influenza virus infection comprising peanut mushroom extract or a compound isolated therefrom.

The present invention relates to a composition for preventing or treating influenza virus infection comprising peanut mushroom extract or a compound isolated therefrom as an active ingredient.

H1N1, H5N1, H3N2, H7N7, and H9N2 viruses have been identified as the major infectious agents of avian influenza viruses and have caused massive human casualties, panic attacks and economic paralysis that have killed more than two million people worldwide during pandemic influenza. The World Health Organization (WHO) calls for international measures (2005, Global Influenza Preparedness Plan).

In Korea, three outbreaks of avian influenza in 2003, 2006 and 2008 led to massive economic losses in the poultry industry and socioeconomic as well as confirmation of the possibility of infection in mammals. In Korea, asymptomatic infections occurred and recently, pandemic influenza Social concerns about the outbreak are rising.

Currently, there are oseltamivir phosphate and inhalant type zanamivir, which are the oral medicines that selectively inhibit viral originated neuraminidase as the only treatment for avian influenza and swine flu. However, Tamiflu, which is used as an oral therapeutic agent, has side effects such as nausea, vomiting, nervous system or mental disorder and may cause resistance virus against Tamiflu. Recently, there have been reports of the emergence of resistant virus against Tamiflu, which is already lethal to the spread of H1N1 virus, and even human-to-human transmission of Tamiflu-resistant virus has been found. Thus, the antiviral agents developed so far show severe side effects, and a great deal of attention must be paid to their applications. In addition, the development of a vaccine has a problem of low efficacy when the virus type of the prevalent virus is not matched with the virus of the vaccine. Therefore, there is an increasing need to develop a new influenza virus agent having excellent infection control effect and excellent stability.

Meanwhile, peanut mushrooms ( Glaziella splendens ) is a mushroom growing on a perennial tree, or nesting in an overgrown form, and is a subspecies belonging to the peanut mushroom (Glaziellaceae) and the peanut mushroom ( Glaziella ). Peanut mushrooms are round or amorphous, round to round, 1 to 4 cm in diameter, slightly elastic when fresh, but hard to dry. It is light yellow or reddish brown, initially smooth, but gradually forms small spots on the surface and sprouts ascent from black to black.

There have been no studies on the use of peanut mushroom and its extracts in domestic and foreign fields. In particular, the inhibitory effect of peanut mushroom extract on influenza activity has not been studied or reported.

In studying natural products and compounds isolated therefrom in order to develop drugs having an excellent effect of preventing or treating influenza infection, the present inventors have found that peanut mushroom extract has excellent effect of inhibiting the activity of neuraminidase enzyme of influenza virus And completed the present invention.

Korean Patent Application No. 10-2000-0044891 Korean Patent Application No. 10-2004-0075381 Korean Patent Application No. 10-2014-0020793 Korean Patent Application No. 10-2014-0007164

It is an object of the present invention to provide a composition for preventing or treating influenza virus infection comprising peanut mushroom extract as an active ingredient.

It is still another object of the present invention to provide a composition for preventing or treating influenza virus infection comprising as an active ingredient a compound represented by any one of the following formulas (1) to (3) isolated from peanut mushroom extract.

[Chemical Formula 1]

(2)

로 구성된 군으로부터 선택된다.Wherein R is independently -OH, -H and < RTI ID = 0.0 >

≪ / RTI >

(3)

In order to achieve the above object, the present invention provides a pharmaceutical composition and a food composition for preventing or treating influenza virus infection comprising peanut mushroom extract as an active ingredient.

The present invention also provides a pharmaceutical composition and a food composition for preventing or treating an influenza virus infection comprising as an active ingredient a compound represented by any one of the following formulas (1) to (3) isolated from peanut mushroom extract.

[Chemical Formula 1]

(2)

로 구성된 군으로부터 선택된다.Wherein R is independently -OH, -H and < RTI ID = 0.0 >

≪ / RTI >

(3)

The peanut mushroom extract of the present invention and the compounds of formulas (1) to (3) isolated therefrom are excellent in the effect of preventing or treating influenza virus infection by inhibiting the activity of neuraminidase enzyme of influenza virus, And the like can be usefully utilized as a pharmaceutical composition and food for.

The present invention relates to peanut mushrooms ( Glaziella splendens ) as an active ingredient. The present invention also provides a composition for preventing or treating influenza virus infection.

The composition comprises a pharmaceutical composition or a food composition.

Hereinafter, the present invention will be described in more detail.

The peanut mushroom extract as an active ingredient of the present invention can be obtained by using extraction and separation methods known in the art. The 'extract' defined in the present invention can be used as a crude extract, a polar solvent extract or a nonpolar solvent extract .

In the present invention, the peanut mushroom extract is collectively referred to as a concentrate obtained by squeezing the peanut mushroom with an appropriate leaching solution and evaporating the leach solution. The peanut mushroom extract may be prepared by diluting the peanut mushroom with the dried powder of the diluted solution, concentrate or extract, .

Further, in the extraction method, methods such as room temperature extraction, hot water extraction, hot water extraction, cold extraction, hot extraction, pressure extraction, reflux cooling extraction or ultrasonic extraction can be used, But is not limited thereto.

The peanut mushroom extract may be further subjected to a drying method after the extraction, and may include, but is not limited to, freeze drying, vacuum drying, hot air drying, spray drying, vacuum drying, foam drying, high frequency drying, .

The extraction solvent for extracting the peanut mushroom extract is not limited thereto, but one or more solvents selected from water, chloroform, alcohols having 1 to 4 carbon atoms, and a mixed solvent thereof may be used. Preferably, the extraction solvent is a mixture of methanol and chloroform Can be every day.

The peanut mushroom extract may be a peanut mushroom fruit body extract or a fraction thereof.

The fraction may be a fraction that is fractionated with hexane, ethyl acetate, or N-butanol, preferably a fraction that is fractionated with ethyl acetate.

In addition, the present invention provides a composition for preventing or treating influenza virus infection comprising as an active ingredient a compound represented by any one of Chemical Formulas 1 to 3 isolated from peanut mushroom extract.

The composition comprises a pharmaceutical composition or a food composition.

[Chemical Formula 1]

(2)

로 구성된 군으로부터 선택되며, 바람직하게는

일 수 있다.Wherein R is independently -OH, -H and < RTI ID = 0.0 >

, Preferably selected from the group consisting of

Lt; / RTI >

(3)

The Peanut Mushroom Extract of the present invention and the compounds of the above Formulas 1 to 3 isolated from the peanut mushroom extract inhibit neuraminidase activity, and the neuraminidase is preferably neuraminidase derived from H3N2, H1N1 or H5N1, It is preferably H3N2-derived neuraminidase. Neuraminidase is an enzyme of the glycoprotein in the viral envelope membrane, which is necessary when the virus enters or exits the cells of the host. Accordingly, the peanut mushroom extract and the compounds of the above formulas (1) to (3) isolated from the peanut mushroom extract inhibit the function of the neuraminidase, and as a result, the virus is prevented from being infected with other cells, thereby preventing or treating influenza infection.

The peanut mushroom extract of the present invention and the serotypes of influenza virus having the inhibitory activity of neuraminidase of the above-mentioned formulas (1) to (3) separated from the peanut mushroom extract include H1N1, H3N2 or H5N1, preferably H3N2, It does not.

The diseases caused by the influenza virus infection include, but are not limited to, flu, cold, sore throat, bronchitis and pneumonia.

The flu includes, but is not limited to, avian influenza, swine flu or swine flu.

The composition of the present invention may contain one or more known active ingredients having an effect of preventing or treating influenza virus infection together with peanut mushroom extract or the compounds of formulas (1) to (3) isolated therefrom.

The composition of the present invention may further comprise a pharmaceutically acceptable additive, wherein pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose Starch glycolate, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, calcium stearate, calcium stearate, White sugar, etc. may be used. The pharmaceutically acceptable additives according to the present invention are preferably included in the composition in an amount of 0.1 to 90 parts by weight, but are not limited thereto.

The composition of the present invention can be administered orally or parenterally in various clinical formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, And suitable formulations known in the art are preferably those described in Remington ' s Pharmaceutical Science (Mack Publishing Company, Easton PA, recently).

The solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like May be included.

The preparation for parenteral administration includes sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

The parenteral administration can be carried out using an external or intraperitoneal injection, intramuscular injection, subcutaneous injection, intravenous injection, intramuscular injection or intra-thoracic injection.

The dosage of the pharmaceutical composition of the present invention may vary depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of disease, The amount of the extract may be 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, and may be administered once a day or several times.

The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of influenza virus infection.

The compounds of the formulas (1) to (3) of the present invention can be used in the form of pharmaceutically acceptable salts and include all salts, hydrates and solvates prepared by conventional methods.

As salts, acid addition salts formed by pharmaceutically acceptable free acids are useful. The acid addition salt is prepared in a conventional manner, for example, by dissolving the compound in an excess amount of an aqueous acid solution and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The same molar amount of the compound and the acid or alcohol (e.g., glycol monomethyl ether) in water may be heated and then the mixture may be evaporated to dryness, or the precipitated salt may be subjected to suction filtration. As the free acid, organic acids and inorganic acids can be used. As the inorganic acids, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid and the like can be used. Examples of the organic acids include methanesulfonic acid, p- toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycollic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid and the like can be used. It does not.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the non-soluble compound salt, and evaporating and drying the filtrate. At this time, as the metal salt, it is preferable to produce sodium, potassium or calcium salt particularly, but not limited thereto. The corresponding silver salt can also be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).

Pharmaceutically acceptable salts of such compounds include, unless otherwise indicated, salts of acidic or basic groups that may be present in the compounds. For example, pharmaceutically acceptable salts may include sodium, calcium and potassium salts of hydroxy groups, and other pharmaceutically acceptable salts of amino groups include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, (Mesylate) and p -toluenesulfonate (tosylate) salt, and the like, and a method for producing a salt known in the art ≪ / RTI >

The term "prophylactic" in the present invention means all the acts of inhibiting or delaying an influenza virus infection by the administration of the pharmaceutical composition of the present invention, and the term " treatment "refers to the symptom caused by an influenza virus infection Means any act that improves or is beneficially modified.

In the present invention, the term "food" refers to a food having a biological control function such as prevention and improvement of disease, bio-defense, immunity, recovery after disease, suppression of aging and the like.

When the peanut mushroom extract of the present invention or the compounds of formulas (1) to (3) isolated therefrom is used as a food additive, the peanut mushroom extract or the compounds of formulas (1) to (3) And can be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). Generally, the peanut mushroom extract of the present invention or the compounds of the general formulas (1) to (3) isolated therefrom are added in an amount of 15% by weight or less, preferably 10% by weight or less based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

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

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharine and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, A carbonating agent used in a carbonated beverage, and the like. In addition, the composition of the present invention may comprise flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally 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.

The term " prophylactic "in the present invention means any action which inhibits or delays influenza virus infection by the administration of the food composition of the present invention, and the term " improvement" refers to the symptoms of influenza virus infection Means any act that improves or is beneficially modified.

Hereinafter, preferred examples, experimental examples, and formulation examples are provided to facilitate understanding of the present invention. However, the following examples, experimental examples and preparation examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited thereto.

Example  1. Preparation of peanut mushroom extract

The present inventors conducted the following experiment to produce peanut mushroom extract having anti-influenza effect from peanut mushroom.

Dried peanut mushroom fruiting bodies were extracted with methanol / chloroform (1: 1) as an extraction solvent at room temperature for 3 days, left to stand at room temperature for 3 days, filtered with filter paper or the like and concentrated to obtain peanut mushroom extract .

Example  2. Peanut mushroom Fraction  Produce

The extract obtained in Example 1 was diluted with water and then sequentially fractionated with the same amount of nucleic acid, ethyl acetate and butanol, and concentrated under reduced pressure to obtain respective solvent fractions. The fractions thus obtained were used as experimental samples for the separation and purification of the compounds 1 to 5.

Example  3. Isolation and Identification of Compounds 1 to 5

The peanut mushroom ethyl acetate fraction obtained in Example 2 was subjected to silica gel column chromatography using chloroform: methanol (100: 1-1: 1, v / v). Subsequently, the fraction obtained by performing Sephadex LH-20 column chromatography using methanol was further purified to obtain Compound 2, and Compound 1 was obtained by preparative-HPLC, and MPLC was performed using 40-100% methanol Compounds 3, 4 and 5 were isolated. The chemical structures of these compounds were confirmed by spectral analysis ( ¹ H NMR, ¹³ C NMR and mass spectrometry), and the results are shown below.

Example  3-1. Identification of Structure of Compound 1

The results of 1 H NMR and ¹³ C NMR spectra of Compound 1 are shown in Table 1.

No. δ _H δ _C One 4.97, 4.79 (d, 12.37) 65.1 2 3 167.4 4 5.36 (s) 104.3 4a 149.3 5 3.02, 2.73 (d, 19.59) 33.0 6 5.65 (t) 78.4 7 75.6 8 197.3 8a 113.9 9 1.40 (s) 23.7 10 2.31 (m) 44.7 11 3.99 (m) 66.7 12 1.17 (d 6.19) 23.5 One' 105.8 2' 164.1 3 ' 6.11 (d, 2.06) 101.8 4' 166.4 5 ' 6.12 (d, 2.06) 112.7 6 ' 145.0 7 ' 2.22 (s) 24.6 8' 172.2

¹ H NMR ( CD ₃ OD ) : In order to confirm the binding relationship of hydrogen per carbon, ¹ H NMR spectra were measured, and two meta-coupled aromatic hydrogens were observed at 6.12 and 6.11 ppm, Methylene hydrogen bonded to oxygen, olefin methine hydrogen at 5.36 ppm, and methylene hydrogen bonded to oxygen at 4.97 and 4.79 ppm were observed. In addition, methine hydrogen bound to oxygen was observed at 3.99 ppm, and three methyl hydrogens were observed at 3.02, 2.73, and 2.31 ppm at methylene hydrogen, 2.22, 1.40, and 1.17 ppm.

¹³ C NMR ( CD ₃ OD ) : In order to confirm the carbon-carbon bond relationship, a ¹³ C NMR spectrum was measured. As a result, carbonyl carbon was observed at 197.3 and 172.2 ppm and 167.4, 166.4, 164.1, 149.3, 145.0, 113.9, 112.7, 105.8, 104.3, and 101.8 ppm, and methine carbon bound to oxygen was observed at 78.4, 75.6, and 66.7 ppm, and methylene carbon bonded at oxygen at 65.1 ppm, 44.7 and 33.0 ppm, and three methyl carbons were observed at 24.6, 23.7 and 23.5 ppm.

Mass spectrum : High resolution ESI mass spectrometry was performed to confirm the molecular weight and molecular formula of the compound, and the result was a measurement of m / z 405.1540 [M + H] ⁺ (-0.9 mmu).

The chemical structure of the novel compound 1, in which the molecular formula is C ₂₁ H ₂₄ O ₈ and the molecular weight is 404, was synthesized as a result of the spectrum data as described above.

Example  3-2. Identification of Structure of Compound 2

¹ H NMR ( CD ₃ OD ) : ¹ H NMR spectrum was measured to confirm the binding relationship of hydrogen per carbon. As a result, olefin methine hydrogen was observed at 6.51 and 6.20 ppm, and 6.13. At 6.12 ppm two meta-coupled aromatic hydrogens were observed, and at 5.58 ppm methine hydrogen bound to oxygen was observed. In addition, double bond hydrogens at 5.51 ppm, methylene hydrogens bound to oxygen at 4.99 and 4.79 ppm, methylene hydrogens at 3.05 and 2.79 ppm, and two methyl hydrogens at 2.22 and 1.42 ppm were observed.

¹³ C NMR ( CD ₃ OD ) : In order to confirm the carbon-carbon bond relationship, a ¹³ C NMR spectrum was measured, and data corresponding to the chemical structure of the following compound 2 was derived.

Mass spectrum : High resolution ESI mass spectrometry was performed to confirm the molecular weight and molecular formula of the compound, and the result was m / z 402.8 [M + H] ⁺ .

The chemical structure of Compound 2 corresponding to Entonaemin A, which has a molecular formula of C ₂₁ H ₂₂ O ₈ and a molecular weight of 402, was synthesized as follows by synthesizing the above-described spectral data.

Example  3-3. Identification of Structure of Compound 3

¹ H NMR ( CD ₃ OD ) : ¹ H NMR spectrum was measured to confirm the binding relationship of hydrogen per carbon. As a result, olefin methine hydrogen was observed at 6.47, 5.98 and 5.41 ppm, and 6.13 and 6.12 ppm Two methacylated aromatic hydrogens, methine hydrogens bound to oxygen at 5.58 ppm, and methylene hydrogens bound to oxygen at 4.99 and 4.76 ppm were observed. In addition, three methyl carbons were observed at 3.06 and 2.77 ppm and at 2.22, 1.84 and 1.41 ppm, respectively.

¹³ C NMR ( CD ₃ OD ) : In order to confirm the carbon-carbon bond relationship, a ¹³ C NMR spectrum was measured, and data corresponding to the chemical structure of the following compound 3 was derived.

Mass spectrum : High resolution ESI mass spectrometry was performed to confirm the molecular weight and molecular formula of the compound, and the measurement was m / z 384.8 [M + H] ⁺ .

The chemical structure of Compound 3 corresponding to Comazaphilone D, whose molecular formula is C ₂₁ H ₂₂ O ₇ and whose molecular weight is 384, was synthesized as follows by synthesizing the above-described spectral data.

Example  3-4. Structure identification of compound 4

¹ H NMR ( CD ₃ OD ) : ¹ H NMR spectrum was measured to confirm the binding relationship of hydrogen per carbon. As a result, olefin methine hydrogen was observed at 6.42, 6.21, and 5.55 ppm, Methine hydrogen bonded to oxygen at 5.58 ppm, and methylene hydrogen bonded to oxygen at 4.99, 4.76 and 4.69 ppm were observed. In addition, three methyl hydrogens were observed at 3.05 and 2.78 ppm at methylene hydrogen, at 2.22, 2.08 and 1.42 ppm, respectively.

¹³ C NMR ( CD ₃ OD ) : In order to confirm the carbon-carbon bond relationship, a ¹³ C NMR spectrum was measured, and data corresponding to the chemical structure of the following compound 4 was derived.

Mass spectrum : High resolution ESI mass spectrometry was performed to confirm the molecular weight and molecular formula of the compound, and the result was a measurement of m / z 444.8 [M + H] ⁺ .

The chemical structure of Compound 4 corresponding to Rubiginosin A, whose molecular formula is C ₂₃ H ₂₄ O ₉ and whose molecular weight is 444, was synthesized as follows by synthesizing the above-described spectral data.

Example  3-5. Structure identification of compound 5

¹ H NMR ( CD ₃ OD ) : ¹ H NMR spectrum was measured to confirm the binding relationship of hydrogen per carbon. As a result, two meta-coupled aromatic hydrogens were observed at 6.13 and 6.12 ppm, Olefin hydrogen, methine hydrogen associated with oxygen at 5.58 ppm was found. Three methyl hydrogens were observed at oxygen concentrations of 5.07 and 4.63 ppm, methyl hydrogen at 3.25 and 3.20 ppm, and methyl hydrogen at 3.04 and 2.78 ppm, respectively.

¹³ C NMR ( CD ₃ OD ) : In order to confirm the carbon-carbon bond relationship, a ¹³ C NMR spectrum was measured, and as a result data corresponding to the chemical structure of the following compound 5 was derived.

Mass spectrum : High resolution ESI mass spectrometry was performed to confirm the molecular weight and molecular formula of the compound, and the result was m / z 402.8 [M + H] ⁺ .

The chemical structure of Compound 5 corresponding to Entonaemin B, which has a molecular formula of C ₂₂ H ₁₀ O ₈ and a molecular weight of 402, was synthesized as follows by synthesizing the above-described spectral data.

Experimental Example  1: peanut mushroom extract H3N2  Neuraminidase ( neuraminidase ) Inhibition activity measurement

In order to confirm the effect of the peanut mushroom extract obtained in Examples 1 and 2 on influenza A virus, the influenza A virus, serotype H3N2-derived neuraminidase (R & D system, 6875) and substrate 4-Methylumbelliferyl- DN-acetylneuraminic acid (SIGMA M8639) was used to conduct experiments to measure inhibitory activity against peanut mushroom extract against neuraminidase.

First, 10 μL of each of the peanut mushroom extracts obtained in Examples 1 and 2 was added, and 50 μL of the substrate at a concentration of 200 μM was added. A 25 mM MES (concentration of 25 mM) containing 5 mM CaCl ₂ and 500 mM NaCl 90 μL of buffer solution (pH 6.5) was added and mixed. After that, 50 μL of neuraminidase with an enzyme concentration of 50 ng / mL was added and reacted. Then, the light absorbance at 365 nm and the luminescence at 445 nm were measured with a fluorescence spectrometer. Thereafter, the inhibitory activity of neuraminidase was calculated as a percentage from the above measurement results by the following [formula 1], and the results are shown in Table 2.

[Equation 1]

Inhibition activity (%) = ((RFU of untreated control group + RFU of sample) / RFU of control group) × 100

Sample name (200 μg / ml) % Inhibition Methanol extract 30.2 Hexane fraction 42.9 Ethyl acetate fraction 74.4 Butanol fraction 17.9

As shown in Table 2, the peanut mushroom methanol extract of the present invention showed 30.2% inhibition activity of neuraminidase enzyme, and it was confirmed that the ethyl acetate fraction showed 74.4% high neuraminidase inhibitory activity Respectively. (In the above, the methanol extract refers to the extract extracted with a mixed solvent of methanol and chloroform.)

This means that the peanut mushroom extract of the present invention can be used for prevention or treatment of influenza virus infection by inhibiting the activity of influenza virus.

Experimental Example  2: Compounds 1 to 5 isolated from peanut mushroom H3N2  Measurement of inhibitory activity of neuraminidase

In order to confirm the effect of the active compounds 1 to 5, which are active substances isolated from the peanut mushroom ethyl acetate fraction isolated in Example 3, on influenza A virus, serine-type H3N2-derived neuraminidase (R & D system, 6875) and the substrate, 4-Methylumbelliferyl-DN-acetylneuraminic acid (SIGMA M8639), were used to measure inhibitory activity against other types of neuraminidase.

First, 10 μL of each of the compounds 1 to 5 obtained in Example 3 was added, and 50 μL of a 200 μM substrate was added. A 25 mM MES buffer solution to which 5 mM CaCl ₂ and 500 mM NaCl were added (pH 6.5) were added and mixed. After that, 50 μL of neuraminidase with an enzyme concentration of 50 ng / mL was added and reacted. Then, the light absorbance at 365 nm and the luminescence at 445 nm were measured with a fluorescence spectrometer. Thereafter, the inhibitory activity of neuraminidase was calculated as a percentage from the above measurement results by the following [formula 1], and the results are shown in Table 3.

Name of sample IC ₅₀ Ki Inhibition form Zanamivir (Relenza) 12.2 (nM) - Competitive inhibition Compound 1 235.8 [mu] M - Non-competitive inhibition Compound 2 243.8 [mu] M - Non-competitive inhibition Compound 3 30.9 [mu] M 35.3 Non-competitive inhibition Compound 4 46.5 [mu] M 42.0 Non-competitive inhibition Compound 5 177.4 [mu] M - Non-competitive inhibition

Compared to Zanamivir, a control group that is known to competitively inhibit the neuraminidase enzyme, the compounds isolated from the peanut mushroom ethyl acetate fraction, as shown in Table 3, inhibit H3N2 neuraminidase enzyme activity in a concentration-dependent manner The IC ₅₀ values of the compounds 1 to 5 were measured to be 235.8 μM (1), 243.8 μM (2), 30.9 μM (3), 46.5 μM (4) and 177.4 μM (5), respectively. In particular, it was confirmed that the compounds 3 and 4 exhibit excellent influenza virus activity inhibitory effects.

This means that the compounds isolated from peanut mushroom extract of the present invention can be used for prevention or treatment of influenza virus infection by inhibiting the activity of influenza virus.

Hereinafter, a pharmaceutical composition for preventing or treating influenza virus infection comprising the peanut mushroom extract and a compound of the present invention as an active ingredient and a preparation example of a food composition for prevention or improvement are described below, but the present invention is not limited thereto, This is specifically intended to explain.

Formulation Example 1. Preparation of pharmaceutical preparations

1. Manufacturing of powder

Peanut mushroom extract or compounds isolated therefrom 20 mg

Lactose 100 mg

Talc 10 mg

The above components can be mixed and filled in an airtight container to prepare powders.

2. Preparation of tablets

Peanut mushroom extract or compounds isolated therefrom 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

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

3. Preparation of capsules

Peanut mushroom extract or compounds isolated therefrom 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components may be mixed and filled in a gelatin capsule according to a conventional method for preparing a capsule, thereby preparing a capsule.

4. Preparation of injections

Peanut mushroom extract or compounds isolated therefrom 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule according to the usual injection preparation method.

5. Manufacture of liquids

Peanut mushroom extract or compounds isolated therefrom 20 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component is added to and dissolved in purified water according to a conventional liquid preparation method, and the lemon flavor is added in an appropriate amount, then the above components are mixed, and then purified water is added. The total amount of the liquid agent added to the purified water is adjusted to 100 ml, and the liquid agent can be finally prepared by filling it in a brown bottle and sterilizing it.

Formulation Example 2. Preparation of food preparation

1. Manufacture of health food

Peanut mushroom extract or compounds isolated therefrom 100 mg

Vitamin mixture quantity

70 g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 g of vitamin B12

Vitamin C 10 mg

Biotin 10 g

Nicotinic acid amide 1.7 mg

Folate 50 g

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

2. Manufacture of health drinks

Peanut mushroom extract or compounds isolated therefrom 100 mg

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 캜 for about 1 hour. The resulting solution was filtered to obtain a sterilized 2L container, sealed sterilized, May be used in the manufacture of health beverage compositions.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (12)

Peanut Mushroom ( Glaziella The present invention relates to a pharmaceutical composition for preventing or treating an influenza virus infection,
The pharmaceutical composition for preventing or treating influenza virus infection according to claim 1, wherein the extract is extracted with water, chloroform, C ₁ -C ₄ alcohol or a mixed solvent thereof.
The pharmaceutical composition according to claim 1, wherein the extract is a peanut mushroom fruit body extract or a fraction thereof.
The pharmaceutical composition according to claim 3, wherein said fraction is fractionated by a solvent selected from the group consisting of hexane, ethyl acetate and N-butanol.
The pharmaceutical composition for preventing or treating influenza virus infection according to claim 1, wherein the extract comprises a compound selected from the following formulas (1) to (3):
[Chemical Formula 1]

(2)

In the formula,
Wherein R is independently -OH, -H and < RTI ID = 0.0 >

≪ / RTI >
(3)

6. The compound according to claim 5, wherein R in the formula (2)

Or a pharmaceutically acceptable salt or solvate thereof.
The pharmaceutical composition according to any one of claims 1 to 6, wherein the composition inhibits neuraminidase activity.
The pharmaceutical composition according to claim 7, wherein the composition inhibits H3N2 neuraminidase activity.
The method according to any one of claims 1 to 6, wherein the disease caused by the influenza virus infection is at least one selected from the group consisting of influenza, cold, sore throat, bronchitis and pneumonia. Or a pharmaceutically acceptable salt thereof.
The pharmaceutical composition according to claim 9, wherein the influenza virus is at least one selected from the group consisting of avian influenza, swine flu and swine flu.
A food composition for preventing or improving influenza virus infection comprising peanut mushroom extract as an active ingredient.
12. The food composition according to claim 11, wherein the extract comprises a compound selected from the following formulas (1) to (3).
[Chemical Formula 1]

(2)

In the formula,
Wherein R is independently -OH, -H and < RTI ID = 0.0 >

≪ / RTI >
(3)