KR20110110381A - Composition for the prevention and treatment of highly pathogenic avian influenza virus containing black ginseng bud processed dry powder or extract thereof as an active ingredient - Google Patents

Abstract

The present invention relates to a composition containing the dried black ginseng bud dry powder or its extract as an active ingredient prepared using a high temperature and high pressure manufacturing method, and in detail, the composition of the present invention is an excellent high pathogenic avian influenza virus (highly pathogenic avian influenza). Since it shows an inhibitory effect on, it can be usefully used as a pharmaceutical composition and health functional food for the prevention and treatment of high pathogenic avian influenza virus.

Description

A composition comprising the dried flower bud powder of Black Panax ginseng or the extract isolated there from for preventing and treating pathogenic avian influenza virus}

It is an object of the present invention to provide a pharmaceutical composition and anti-viral composition for antiviral, which has an inhibitory effect on a highly pathogenic avian influenza virus containing a dry powder or extract of black ginseng bud prepared using a high temperature and high pressure manufacturing method as an active ingredient. will be.

Selmons et al., Avian . Dis ., 18 (1) , pp. 119-124, 1974.

2 Webster RG et al., Microbiol . Rev. , 56 (1) , pp. 152-179, 1992.

Document 3 Alexander DJ, Vet. Microbiol . , 74 (1-2) , pp. 3-13, 2000.

[4] Song CS et al., Korean J. Poult . Sci ., 31 (2) , pp. 129-136, 2004.

Document 5 JH, Bull. NY Acad . Med . , 54 , pp. 855-864, 1978.

[6] Taubenberger JK et al., Science , 275 , pp. 1179-1796, 1997.

7 Potter CW, J. appl . Microbiol ., 91 , pp. 572-579, 2001.

[Document 8] Oxford JS, Rev. Med . Virol ., 10 (2) , pp. 119-133, 2000.

9 Suarez DL et al., J. Virol . , 72 (8) , pp.6678-6688, 1998.

Document 10 Capua I et al., Avian Pathol ., 32 , pp. 47-55, 2003.

Document 11 Capua I and Marangon S, Avian Pathol . , 32 , pp.335-343, 2003.

12 Swayne DE et al., Avian Dis . , 41 , pp. 910-922, 1997.

13 Swayne DE et al., Avian Dis . , 44 , pp. 132-137, 2000.

Ward P et al., J. antimicrob . Chemother ., 55 ( supp1 ) , pp.i5-i21, 2005.

[Reference 15] Understanding Korean Ginseng , Korean Ginseng Society , p.9, 1995.

[Reference 16] Advances in Ginseng Research, Korean Ginseng Society , pp. 127-137, 1998.

[Ref. 17] Comparison of Saponin Contents of Root, Leaf, and Pear of Ginseng , Korean Ginseng Society , p.118, 1987.

[Document 18] Republic of Korea Patent Registration 10-2008-0844991.

Influenza virus is an RNA virus belonging to the Family Orthomy xoviridae, and serotypes are classified into three types, type A, type B and type C. Types B and C have been identified only in humans, and type A has been identified in humans, horses, pigs, other mammals, and various types of poultry and wild birds (Selmons et al., Avian) . Dis ., 18 (1) , pp. 119-124, 1974; Webster RG et al., Microbiol Rev., 56 (1) , pp. 152-179, 1992).

The serotypes of influenza A virus are classified according to the two types of proteins on the surface of the virus, hemagglutinin (HA) and neuraminidase (NA), and 144 types (HA type) 16 species and 9 NA types). The HA protein plays a role in attaching the virus to somatic cells, and the NA protein allows the virus to penetrate into the cell (Alexander DJ, Vet. Microbiol . , 74 (1-2) , pp.3-13, 2000). The normal natural host of influenza A virus is known as wild waterfowl such as ducks, seagulls, etc .. As a result of epidemiological investigations of influenza infection in wild birds around the world, all 16 existing HA and 9 NA types Influenza viruses have been found to infect wild birds (Selmons et al., Avian) . Dis . , 18 (1) , pp. 119-124, 1974). Avian influenza virus, classified as type A, is a common infectious disease virus, a non-pathogenic avian influenza that causes mild respiratory symptoms when infected with chickens, and low pathogenicity that causes mortality and spawning degradation of about 1 to 30%. Low pathogenic avian influenza (LPAI) and high pathogenic avian influenza (HPAI), which are more than 95% highly deadly and also called bird flu, are classified into three types. (Alexander DJ, Vet . Microbiol ., 74 (1-2) , pp. 3-13, 2000). The highly pathogenic avian influenza is classified as Class A by the International Water Services Bureau (OIE) and domestic type 1 epidemics.

High pathogenic avian influenza (HPAI) has been reported since 1980 in the United States (1983), Australia (1985, 1992, 1994, 1997), Mexico (1994), Pakistan (1994, 2004), Hong Kong (1997, 2001), It has been confirmed worldwide, in Italy (1997, 1999), the Netherlands (2003), Belgium (2003), Germany (2003) and Canada (2004). In particular, in December 2003, serotype A / H5N1 HPAI occurred almost simultaneously in all of Southeast Asia and Far East Asia, including Vietnam, Japan, Thailand, Cambodia, Laos, Laos, Indonesia, and China. In particular, HPAI, which originated in Vietnam and Thailand, is a variant A / H5N1 HPAI virus that can infect human body when it comes into contact with infected birds, unlike the traditional HPAI that does not infect humans in Korea and Japan at that time. By March 2004, 22 people in contact with infected birds were infected in Vietnam, and 15 of them were killed. In neighboring Thailand, 11 people in contact with infected birds were infected, and eight of them were killed. It is true. In recent years, the incidence of HPAI has increased by 10 times compared to the past, and especially since 2001, it is taking place every year in the world. Therefore, a new concept of preventive measures that can effectively control HPAI is required. CS et al., Korean J. Poult . Sci . , 31 (2) , pp . 129-136 , 2004).

The first record of human influenza dates back to 412 B.C.E., but the first pandemic influenza record of humanity is estimated to have occurred throughout Europe from 1173 to 1174. Since the 20th century, records of influenza have been more scientifically treated, based on the data that have been left behind, since the 20th century there have been three pandemic influenza. Since the twentieth century, a worldwide pandemic between 1918 and 1920, the first influenza pandemic (aka Spanish flu) has been documented as the greatest damage to humanity, with between 20 and 50 million people dying during that period ( Walter JH, Bull. NY Acad . Med . , 54 , pp. 855-864, 1978). The causative agent of the Spanish flu virus has been identified as serotype A / H1N1, which is very similar to the swine influenza virus and is still a major epidemic of pandemic flu, which is still prevalent worldwide every year (Taubenberger JK et al., Science , 275 , pp.1793-). 1796, 1997). Since the 20th century, which occurred between 1957 and 1958, the second influenza pandemic began in China and rapidly spread along the coast to nearby Hong Kong, Singapore, Japan and Taiwan in six to seven months. About 40-50% of the world's population was infected during this period, of which 25% had clinical symptoms, mainly infants and middle-aged people who died of infection, and about 1 million people died. (Potter CW, J. appl . Microbiol ., 91 , pp. 572-579, 2001). The causative virus was identified as serotype A / H2N2 influenza virus. In addition, the third pandemic of the influenza pandemic between 1968 and 1969 was confirmed to originate in Hong Kong and spread rapidly to Taiwan, the Philippines, Singapore and Vietnam. The causative virus is serotype A / H3N2, which differs from the previous serotype H2N2 virus and HA, which has been analyzed to be transmitted from birds, and is still the main epidemic of pandemic flu, which is still prevalent worldwide every year. (Oxford JS, Rev. Med. Virol., 10 (2), pp.119-133, 2000).

Some serotypes of AIV, a major concern in birds, are infected by humans, causing flu and causing death. Several strains of avian-induced avian influenza viruses, including serotype A / H5N1, which are now called Hong Kong Bird Flu, are likely to infect humans. It is assumed that there is. Therefore, the present study of the mutant viruses derived from these three serotypes is being conducted worldwide (Suarez DL et al., J. Virol . , 72 (8) , pp.6678-6688, 1998). .

The case of human infection that overcomes the barriers between avian influenza virus (serum-type A / H5N1 avian influenza virus) recently reported in Vietnam is the prelude to the pandemic of the fourth human influenza pandemic since the 20th century. Attention has been paid to the spread of the mutant virus, including direct transmission from person to person. The mutant virus is an avian influenza virus belonging to serotype A / H5N1, which is also avian influenza virus that has been prevalent only in birds for the first time among humans.

Influenza viruses can infect the respiratory tract, cause systemic symptoms, change their appearance periodically, and move to other hosts without killing them before they die. It is the virus that causes the greatest economic loss to humans, and although the vaccine has been developed, it has not caught up with the mutation of the virus, and there is no fundamental virus treatment yet.

Among the avian influenza vaccines, live virus vaccines are almost impossible to develop due to the characteristics of viruses that are easily mutated. The vaccines developed to date can be largely classified into a deadly poison vaccine and a recombinant vaccine. As HPAI outbreaks prolonged and spread throughout the country in 1999 and Hong Kong in 2003, avian influenza vaccine was used as a means of combating HPAI in combination with a selective killing policy. Currently, in Italy and Hong Kong, the use of avian influenza vaccine It is positively regarded as effective in controlling the disease (Capua I et al., Avian Pathol . , 32 , pp. 47-55, 2003; Capua I and Marangon S, Avian Pathol ., 32 , pp. 335-343, 2003).

Vaccines positively evaluated in Italy (serum type A / H7N1) and Hong Kong (serum type A / H5N1) are all serotoxin vaccines and are heterologous serotypes with the same HA type but different NA type (serum type A / H7N3, A serotoxin vaccine with serotype A / H5N2) was used to distinguish it from field infection in antibody testing. However, the deadly venom vaccine cannot be distinguished from the vaccine antibody and the outdoor infection by the Agar Gel Precipitation (AGP) test, which is the standard type A bird influenza standard diagnosis. It is pointed out that it is not suitable for inspection. In addition, the currently developed dead venom vaccine can reduce the amount of virus released into the feces during HPAI infection, but does not completely prevent the spread of the disease (Capua I et al., Avian Pathol ., 32 , pp . 47-55, 2003; Capua I and Marangon S, Avian Pathol., 32 , pp. 335-343, 2003).

In Mexico, when the serotype A / H5N2 low-pathogenic avian influenza developed in late 1993, the virus spread throughout the country became highly pathogenic and abolished pathogenicity. The vaccine is produced and used, and has been converted to advanced genetic vaccination to date to distinguish between the vaccination system and the field infection system. The vaccines used in Mexico so far include the hemagglutin gene, which is the surface protein of HPAI virus, in fowl pox virus, which is an advanced genetic recombinant vector that can effectively prevent chickenpox and HPAI at the time of vaccination. Vaccines, and on the other hand, countries that use chicken pox vaccines, including Korea, have been evaluated to have a disadvantage in that the effects of viral vector vaccines are halved due to the interference of antibodies induced by conventional poultry vaccination. In addition, genetically engineered viral vector vaccines have been developed by inserting the Hemagglutin gene, a surface protein of HPAI virus, into infectious laryngotracheitis virus (ILT). In the countries of use, the effect is halved, and it is estimated that the only available target is chickens (Swayne DE et al., Avian) . Dis . , 41 , pp. 910-922, 1997; Swayne DE et al., Avian Dis . , 44 , pp. 132-137, 2000).

Amantadine and rimantadine are substances that inhibit the function of M2 ion channel proteins of influenza viruses and are representative antiviral agents that inhibit the growth of influenza viruses in vivo. However, these two antiviral agents were found to be effective only against serotype A influenza virus and not against serotype B influenza virus without M2 protein. In addition, amantadine and rimantadine have been found to have the disadvantage that the emergence of a mutant virus that does not affect the ion channel function of influenza virus M2 protein when used. Zanamivir and oseltamivir, which are developed to compensate for these drawbacks, inhibit the function of the neuraminidase protein of influenza viruses and are representative of the suppression of influenza virus proliferation in vivo. Antiviral agents. These two antiviral agents are known to be effective against all 16 serotype A influenza viruses and serotype B influenza viruses. However, zanamivir has the disadvantage of inhalation and intravenous administration, while oseltamivir can be administered orally, but it has been pointed out as a disadvantage due to side effects such as recent reports of resistant virus and vomiting and dizziness upon oral administration (Ward P et. al.,. J. antimicrob. Chemother, 55 (supp1), pp.i5-i21, 2005). Therefore, along with vaccines and therapeutics, the development of safe natural materials while increasing human immunity will be an important means of reducing mortality during pandemic.

Ginseng is a perennial plant belonging to the genus Panax, and the genus Panax is a perennial root of the genus Araliaceae. Typical species include Panax ginseng, Panax quinquefolia, Panax notoginseng, Panax japonica, Panax trifolia, Panax pseudoginseng, Panax pseudoginseng, and Panax ginseng. vietnamensis) (Understanding Korean Ginseng , Korean Ginseng Society , p.9, 1995; Advances in Ginseng Research, Korean Ginseng Society , pp. 127-137, 1998).

The most important component of these Panax plants is saponin. In particular, the plant of the genus Panax, unlike other plants contain a common saponin 1 to 4 sugars are bonded to the dammarane skeleton. In particular, saponins with high content in Korean ginseng are ginsenosides Rb1, Rb2, Rc, Rd, Rg1 and Re. These saponin components show various effects, and the types and strengths of the drugs vary greatly depending on their structure (Understanding Korean Goryeo Ginseng Society , Korean Ginseng Society , p.9, 1995).

However, the research on ginseng and processed ginseng so far is mostly limited to the roots. In fact, the main effect of ginseng saponin indicates the presence of 4-5% in the ginseng roots and buds are known to exist around four times higher than 18-20%, but (the roots of ginseng leaf and compare tribute saponin, ginseng Society , p.118, 1987) Very little research has been conducted on ginseng buds. Therefore, in this laboratory, as a result of further efforts to increase the active ingredient content of ginseng, the principle of Gujeungpo, which is the most representative method, is to use water and fire together in the processing method of Chinese herbal medicine, which is one of steaming methods. Patents have been filed for the preparation of standardized black ginseng buds that can be shortened and mass-produced, and for anti-aging compositions of sub extracts (Korean Patent Application No. 10-2006-0113815).

However, there is no disclosure or teaching on the inhibitory effect on the HPAI of black ginseng bud dry powder or its extract prepared by the method disclosed in Korean Patent Application No. 10-2006-0113815.

Accordingly, the present inventors have completed the present invention by confirming that the black ginseng bud dry powder prepared by the high temperature and high pressure method or its extract exhibits an excellent inhibitory effect on the highly pathogenic avian influenza virus in the era of premiumization of ginseng.

In order to achieve the above object, the present invention is a pharmaceutical for the prevention and treatment of high pathogenic avian influenza virus containing a dry powder or extract thereof of Black Panax Ginseng flower bud prepared by high temperature and high pressure manufacturing method as an active ingredient. To provide a composition.

In addition, the present invention provides a health food for the prevention and improvement of high pathogenic avian influenza virus containing a dry powder of the black ginseng bud prepared by the method of producing high temperature and high pressure or its extract as an active ingredient.

The dry powder as defined herein is characterized in that black ginseng buds are naturally dried, freeze dried or hot air dried, preferably hot air dried.

 The extract defined herein is characterized in that the crude extract of the black ginseng bud or the polar solvent soluble extract.

The crude extract as defined herein includes crude extracts available in water, including purified water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, preferably water, ethanol or mixed solvents thereof.

The polar solvent soluble extract as defined herein includes water containing purified water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, preferably extracts soluble in water, butanol or a mixed solvent thereof.

The highly pathogenic avian influenza virus as defined herein is H1N1, H1N2, H2N2, Human B, H3N2, H3N8, H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N2, H7N3, H7N4, H7N7, H9N7, H9N2 , H3N2, Human B, H5N1, H9N2, H7N1 or has a serotype of H7N2.

Hereinafter, the present invention will be described in detail.

Dry powder of the black ginseng bud of the present invention and its extract may be prepared through the following process.

For example, the present invention, after washing the buds of about 2 to 7 years old, preferably about 3 to 6 years old ginseng, 12 to 36 hours, preferably at a temperature of 25 to 65 ℃, preferably 35 to 55 ℃ Preferably a first drying step of drying for 18 to 28 hours; The dried ginseng bud is 3 to 9 hours, preferably at a temperature of 95 to 155 ° C., preferably 110 to 145 ° C. under a pressure of 0.05 to 0.45 MPa, preferably 0.10 to 0.20 MPa, more preferably 0.10 to 0.14 MPa. Preferably a first steaming step of steaming for 4 to 7 hours; Shortening the time, including a second drying process step of drying for 8 to 16 hours, preferably 10 to 14 hours at a temperature of 40 to 80 ℃, preferably 50 to 70 ℃, the moisture content within 14% It can be produced through the method of manufacturing a black ginseng bud dry powder (hereinafter referred to as BGF) using high temperature and high pressure that can be mass-produced.

In addition, the crude extract of the present invention, after pulverizing the dry powder obtained above, by adding water of about 1 to 5 times, preferably 2 to 4 times the mass of black ginseng bud dry powder (BGF), 0 to 50 ℃ , Preferably at room temperature for about 3-7 hours, and then a volume of 1-20 times the volume of the solution, preferably 10-15 times the volume of water and ethanol mixed solvent, preferably 50-90% water and ethanol By adding a mixed solvent, 1 to 5 times, preferably about 3 times cold extraction, hot water extraction, ultrasonic extraction, reflux extraction, heating extraction, preferably reflux extraction for 1 to 6 hours, preferably 2 to 4 hours After cooling to room temperature, the ethanol was removed using a vacuum concentrator, and then the crude extract of black ginseng bud of the present invention (hereinafter referred to as “BGFE”) can be obtained.

In addition, the black ginseng bud polar solvent soluble extract of the present invention is about 20 to 30 times the dry weight of the ethanol crude extract of the crude extract, preferably black ginseng bud, preferably 25 to 28 times the volume (v / v). After suspension in water of), fractionation is performed with ether, hexane, ethyl acetate, chloroform, methylene chloride and the like in a nonpolar solvent, preferably in an ether solvent to remove the nonpolar solvent soluble fraction, and the same amount of the remaining water suspension By adding a polar solvent such as butanol and extracting fractions about 1 to 5 times, preferably 2 to 4 times, a black ginseng bud polar soluble extract soluble in a polar solvent such as butanol and water containing a large amount of saponin components can be obtained. have.

The black ginseng bud dry powder prepared by the above method or its extract is present in the active ingredients (Rk1, Rg5, Rg3 (S), Rg3 (R), Rh2) which are not present in the fresh ginseng, white ginseng, or red ginseng, or present in trace amounts. Increased content, specifically, Rb1 0.19-0.21 mg, Rb2 1.05-3.2 mg, Rc 5.4-6.2 mg, Rd 6.6-7.9 mg, Re + Rg1 0.6-1.4 mg, Rf1 per g 0.5-1.1 mg, Rk1 + Rg5 is 76-115 mg, Rg3 (S) + Rg3 (R) is 26-39 mg, Rk3 16-18 mg, Rh4 21-25 mg, Rg4 34-45 mg, Rg6 is 32 Ginsenoside SF as -37 mg and novel substance is characterized by the presence in the range of 0.08-0.11 mg.

In addition, black ginseng bud dry powder or its extract prepared by the production method of the present invention is characterized by exhibiting a strong inhibitory effect on high pathogenic avian influenza virus.

In another aspect, the present invention provides a pharmaceutical composition for the prevention and treatment of high pathogenic avian influenza virus containing the dried powder or extract of the black ginseng bud obtained by the production method as an active ingredient.

The pharmaceutical composition containing the dry powder or extract thereof of the present invention comprises 0.1 to 50% by weight of the dry powder or extract thereof based on the total weight of the composition.

However, the composition is not limited thereto, and may vary depending on the condition of the patient, the type of disease, and the progress of the disease.

Dry powder of the present invention or the extract itself is almost no toxicity and side effects, so it is a drug that can be used with confidence even for long-term administration for the purpose of prevention.

The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.

The compositions of the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Carriers, excipients and diluents that may be included in the composition comprising extracts include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, 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 preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the composition of the present invention is preferably administered at 0.2 to 200 mg / kg, preferably 2 to 100 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The composition of the present invention may be administered to mammals such as rats, mice, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

Black ginseng bud dry powder or extract thereof of the present invention can be used as a raw material with enhanced efficacy than the conventional ginseng has been used, that is, various pharmaceutical preparations, Chinese medicine, health food, food, tea, cosmetics and the like.

In addition, the present invention provides a health food for the prevention and improvement of high pathogenic avian influenza virus containing the dried powder or black extract of black ginseng bud obtained by the above method as an active ingredient.

The composition comprising the dry powder of the present invention or its extract may be used in a variety of drugs, foods and beverages for the prevention and improvement of high pathogenic avian influenza virus. Examples of the food to which the dry powder or extract thereof may be added include various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and are powders, granules, tablets, capsules or beverages. Available in form.

The amount of dry powder or extract of the black ginseng bud in the food or beverage of the present invention is generally added to 0.01-15% by weight of the total food weight of the health food composition of the present invention, the health beverage composition based on 100 ml It can be added at a ratio of 0.02 to 5 g, preferably 0.3 to 1 g.

The health functional beverage composition of the present invention contains the dried black powder or extract of black ginseng bud as an essential ingredient in the indicated ratio, and there is no particular limitation on the liquid component, and various flavors or natural carbohydrates are added as in general drinks. It may contain as a component. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrginine, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 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, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention by confirming that the dried black ginseng bud dry powder or its extract produced by using the high temperature and high pressure manufacturing method exhibits the inhibitory effect on the high pathogenic avian influenza virus, pharmaceutical composition and health functional food for the prevention and treatment of the high pathogenic avian influenza virus It can be used to.

Hereinafter, the present invention will be described in detail with reference to the following examples and experimental examples.

However, the following examples and experimental examples are illustrative of the present invention, and the content of the present invention is not limited by the following examples and experimental examples.

Example  One. Black buds  Produce

After purchasing 1 Kg of 5-year-old ginseng flower buds harvested in May-June grown in Geumsan, it was put in an ultrasonic cleaner (JAC, 4020) and washed three times in total for 3 minutes. The washed ginseng buds were first dried at 50 ° C. for 18 hours using a dryer (Lab Tech-one, LTO-DO-150S) to obtain 434 g.

Take 400 g of the first dried ginseng buds and steam them for 6 hours in a steamer (Sanyo, MLS-3780) for 4 hours at 115 to 135 ℃ under 0.12 MPa, and then heat the internal temperature of the dryer with a built-in hot wire to 60 ℃. 396 g of black ginseng bud (hereinafter referred to as BGF) of the present invention, which was changed to black having a water content of 14% or less by drying for 12 hours in a furnace dryer, was prepared.

Example  2. Black Ginseng Bud  Preparation of Extract

100 g of BGF, black ginseng buds, prepared by the method of Example 1 above, were pulverized with a particle size of 50 to 200 μm by a grinder (Hanil Electronics, HMF-1000A), and 200 ml of water was added thereto for 5 hours at room temperature. After standing, 1.5 L of 80% ethanol (20% water) was added and refluxed three times for 3 hours. The extracted solution was cooled to room temperature and filtered, and then the ethanol was removed using a reduced pressure concentrator. 37.67 g (hereinafter referred to as BGFE) was used as a sample of the virus inhibitory activity of Experimental Example 1 and a sample of acute toxicity test of Experimental Example 2 below.

Example  3. Black Ginseng Bud Crude saponin  Ingredient analysis

500 ml of water was suspended in the black ginseng bud extract obtained in Example 2, and 500 ml of ether was added to remove the nonpolar material. 500 ml of saturated butanol was added to the remaining aqueous layer, followed by extraction of saponin present in ginseng four times, followed by removal of butanol with a reduced pressure concentrator to obtain a butanol extract (crude saponin extract) containing a large amount of ginseng saponin to obtain the saponin component of Experimental Example 1. Used as a sample for analysis.

Experimental Example  1. Saponin ingredient analysis

In order to analyze the components of ginseng saponin present in the black ginseng bud extract, 20 mg of the crude saponin extract obtained in Example 3 was dissolved in 1 ml of methanol, and the filtrate was filtered with a 0.45 μm filter. The components were analyzed at.

As a result of analyzing the components, Rb10.19-0.21 mg, Rb2 1.05-3.2 mg, Rc 5.4-6.2 mg, Rd 6.6-7.9 mg, Re + Rg1 0.6-1.4 mg, Rf1 0.5-1.1 mg, Rk1 + Rg5 76-115 mg, Rg3 (S) + Rg3 (R) 26-39 mg, Rk3 16-18 mg, Rh4 21-25 mg, Rg4 34-45 mg, Rg6 32-37 mg and novel substances Ginsenoside SF was present as 0.08-0.11 mg.

Column: Shiseido C18 column (5, 1504.6 mm) Flow rate 1 ml / min Detector ELSD-LT detector Solvent A; CH ₃ CN: H ₂ O: 5% CH ₃ COOH = 15: 80: 5, B; CH ₃ CN: H ₂ O = 80:20
0 minutes (0% B), 0-10 minutes (30% B), 10-25 minutes (50% B), 25-40 (100% B), 40-50 minutes (100% B), 50-55 Minute (0% B), 55-58 minutes (0% B) Temperature 40

Experimental Example  2. Measurement of antiviral activity

In order to measure the virus inhibitory effect of the ethanol extract of black ginseng bud obtained in the above example, the experiments were performed as follows by applying the method disclosed in the literature.

In this experiment, H5N1 (NIBRG-14; UK NIBSC), a highly pathogenic avian influenza virus, was used. Each experimental group was used for the experiment in the same group of 10 BALB / c mice as a group. Three-week old BALB / c mice (males) purchased from Samta Co. were administered BGFE at a concentration of 50 mg / Kg / day for 30 days. After 30 days, mice were inoculated with lethal H5N1 avian influenza (10 MLD / 50), and mortality was observed daily for 14 days. The control group is the group infected with the virus without administration of the black ginseng bud ethanol extract (BGFE) of the present invention.

sample Dose (mg / kg / day) Lethality (%) BGFE 50 50 Control group - 100

As a result of the experiment, as shown in Table 2, the control group showed 100% mortality after 2 weeks of virus infection. On the other hand, the black ginseng bud ethanol crude extract group (BGFE) showed 50% mortality after 5 of 10 mice died in the experiment 2 weeks after virus infection. That is, black ginseng bud was confirmed to have excellent virus inhibitory effect.

Experimental Example  3. Acute Toxicity Test

Acute toxicity test was performed using 6-week-old specific pathogen-free (SPF) SD rats from the Korea Experimental Supply Center.

After two oral administrations of the BGFE of Example 2 at a dose of 1 g / kg, two animals in each group were observed for mortality, clinical symptoms, and weight changes of the animals. Necropsy was performed to visually observe the abnormalities of the tonic and thoracic organs.

As a result, there were no clinical symptoms or deaths in all animals treated with the test substance, and no toxic changes were observed in weight change, blood test, blood biochemical test and autopsy findings. As a result, the extract of the present invention did not show a change in toxicity even in rats up to 1 g / kg, respectively, it was confirmed that the minimum lethal dose (LD ₅₀ ) is determined to be a safe substance more than 1 g / kg.

The black ginseng bud dry powder of the present invention or an example of the pharmaceutical composition comprising the extract thereof is only to illustrate as follows, and is not intended to limit the present invention by way of example only to be described in detail.

Formulation example  One. Powder  Produce

BGFE 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation example  2. Preparation of Tablets

BGFE 10 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

Formulation example  3. Manufacture of capsule

BGFE 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4. Preparation of injections

BGFE 10 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO ₄ 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation example  5. Liquid  Produce

BGFE 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method for preparing a liquid, each component is added and dissolved in purified water, lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by adding purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation example  6. Manufacture of health food

BGLEB 1000 mg

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

0.13 mg of vitamin B ₁

0.15 mg of vitamin B ₂

0.5 mg of vitamin B ₆

Vitamin B ₁₂ 0.2 g

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

The composition ratio of the above-mentioned vitamin and mineral mixture is mixed with a composition suitable for a health food in a preferred embodiment, but the compounding ratio may be arbitrarily modified. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

BGL 100 mg

15 g of vitamin C

100 g of vitamin E (powder)

Iron lactate 19.75 g

3.5 g of zinc oxide

Nicotinamide 3.5 g

0.2 g of vitamin A

Vitamin B ₁ 0.25 g

Vitamin B ₂ 0.3 g

Water quantification

After mixing the above components in accordance with the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilized and stored in the refrigerator and then Used to prepare the healthy beverage composition of the invention.

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 (8)

After washing the buds of about 2 to 7 years old ginseng, the first drying step of drying for 12 to 36 hours at a temperature of 25 to 65 ℃; Primary steaming step of drying dried ginseng buds for 3 to 9 hours at a temperature of 95 to 155 ° C. under a pressure of 0.05 to 0.45 MPa; Highly pathogenic avian influenza virus containing as an active ingredient the black ginseng bud dry powder obtained by the manufacturing method comprising a second drying process step of drying for 8 to 16 hours at a temperature of 40 to 80 ℃ to dry within 14% moisture content Pharmaceutical compositions for the prevention and treatment of
Dry black powder of black ginseng bud according to claim 1 After pulverizing the dry powder obtained above, about 1 to 5 times the mass of black ginseng bud dry powder (BGF) is added, and it is about 3 to 7 hours at 0-50 degreeC. After being left for a while, 1 to 20 times the volume of the mixed solution of water and ethanol was added, 1 to 5 times reflux extraction was carried out for 1 to 6 hours, cooled to room temperature, filtered and the ethanol was reduced using a vacuum condenser. A pharmaceutical composition for the prevention and treatment of high pathogenic avian influenza virus, which contains the crude extract of black ginseng bud obtained by a manufacturing method comprising the step of removing it as an active ingredient.
After suspension in water of about 20 to 30 times the volume (v / v) of the dry weight of the black ginseng bud crude extract according to claim 2, fractions are performed with ether, hexane, ethyl acetate, chloroform, methylene chloride, and the like in a nonpolar solvent. The non-polar solvent soluble fraction was removed, and the remaining water suspension was added with the same amount of a polar solvent such as butanol and extracted for about 1 to 5 times to obtain a highly pathogenic compound containing a black soybean bud polar solvent soluble extract containing a large amount of saponin as an active ingredient. Pharmaceutical composition for the prevention and treatment of avian influenza virus. The method of any one of claims 1 to 3, wherein the highly pathogenic avian influenza virus is H1N1, H1N2, H2N2, Human B, H3N2, H3N8, H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N2, H7N3, H7N3 Pharmaceutical composition, characterized in that it has a serotype of H7N7, H9N2 or H10N7.
After washing the buds of about 2 to 7 years old ginseng, the first drying step of drying for 12 to 36 hours at a temperature of 25 to 65 ℃; A primary steaming step of drying dried ginseng buds for 3 to 9 hours at a temperature of 95 to 155 ° C. under a pressure of 0.05 to 0.45 MPa; Highly pathogenic avian influenza virus containing as an active ingredient the black ginseng bud dry powder obtained by the manufacturing method comprising a second drying process step of drying for 8 to 16 hours at a temperature of 40 to 80 ℃ to dry within 14% moisture content Functional foods for the prevention and improvement of
      Dry black powder of black ginseng bud according to claim 5 After pulverizing the dry powder obtained above, about 1 to 5 times mass water of black ginseng bud dry powder (BGF) is added, and it is about 3 to 7 hours at 0-50 degreeC. After being left for a while, 1 to 20 times the volume of the water and ethanol mixed solvent was added, 1 to 5 times reflux extraction for 1 to 6 hours and cooled to room temperature, filtered and then ethanol using a vacuum condenser Health functional food for the prevention and improvement of high pathogenic avian influenza virus containing black ginseng bud crude extract obtained by the manufacturing method comprising the step of removing as an active ingredient.       After suspension in water of about 20 to 30 times the volume (v / v) of the dry weight of the black ginseng bud crude extract according to claim 6, fractions are performed with ether, hexane, ethyl acetate, chloroform, methylene chloride, and the like in a nonpolar solvent. The non-polar solvent soluble fraction was removed, and the remaining water suspension was added with the same amount of a polar solvent such as butanol and extracted for about 1 to 5 times to obtain a highly pathogenic compound containing a black soybean bud polar solvent soluble extract containing a large amount of saponin as an active ingredient. Health functional food for the prevention and improvement of avian influenza virus. The method of any one of claims 5 to 7, wherein the highly pathogenic avian influenza virus is H1N1, H1N2, H2N2, Human B, H3N2, H3N8, H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N2, H7N3, H7N3 Functional foods, characterized in that having a serotype of H7N7, H9N2 or H10N7.