KR101363111B1 - Pharmaceutical composition containing paddy husk and barley husk extract as activeingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition comprising a rice hull and barley husk extract as an active ingredient, and more particularly, in the treatment of poultry, fish and livestock exposed to diseases caused by a number of pathogens such as viruses and bacteria. It relates to an effective pharmaceutical composition.
The pharmaceutical composition of the present invention is effective in the treatment of fish and livestock, including a large number of poultry diseases caused by viruses, to protect humans from infection caused by pathogens such as viruses, bacteria, parasites, etc. It can be maintained as, and has anticancer activity, immune enhancing and cholesterol inhibitory effect.

Description

Pharmaceutical composition containing paddy husk and barley husk extract as activeingredient}

The present invention relates to a pharmaceutical composition comprising a rice hull and barley husk extract as an active ingredient, and more particularly, in the treatment of poultry, fish and livestock exposed to diseases caused by a number of pathogens such as viruses and bacteria. It relates to an effective pharmaceutical composition.

Plants have long been a source of various natural substances that help human well-being, and in particular the leaves, bark and roots of some specific plants have been used to treat a variety of diseases and infections. There are thousands of plants that are widely used for therapeutic purposes in developing countries as well as in developed countries.

Most of these substances are secondary metabolites and are used for pharmaceuticals, food additives, cosmetic raw materials, fragrances, and pigments. Despite the synthesis of numerous new compounds by the development of the field of organic synthesis, plants are still recognized as a source of new substances. Considering that there are thousands of new substances with new chemical structures that are found in plants every year, many of them are biologically active, making plants an important ingredient.

Many chemicals produced by plants are classified as primary metabolites because they are essential for cellular function. Primary plant metabolites are universally present in the plant population and comprise most of the chemicals produced by plants, but other compounds are classified as secondary metabolites and are the focal point of both scientific research and commercial use because of their many uses. Has become.

On the other hand, rice (Oryza sativa L.) is an annual herb, although there are many kinds of rice, but it is divided into types of rice, field rice, and waxy rice according to the viscosity of rice grains (米 粒). Rice contains about 75% starch, about 8% gray matter, and 0.5-1% fat. In addition, it contains a small amount of vitamin B. The vitamin content depends on the type of rice and the plantation. Fatty parts include ester and free cholesterol, brassicasterol, stigmasterol, sitosterol, monoglycerides, diglycerides, triglycerides, phospholipids, Free fatty acids and the like. Its pharmacological effects include Bojungikgi (補中益氣), Jebungal (除 煩渴), Jisari (止 瀉痢) efficacy.

Barley (Hordeum vulgare var. Hexastichon) is one of the major cultivated crops, which is known to be good for hangovers.For prevention and treatment of hangovers, brown barley common barley is selected, and whole barley or barley mixed with barley is always recommended. Shin Jae-yong, consent folk remedies, 116-117). Barley contains polyphenolic substances, and these compounds are more contained in the outer layer and embryonic part containing the bark than the inner endosperm of barley, and the barley extract has high antioxidant and active oxygen scavenging ability. Reported. In addition, barley is known to be effective in suppressing cholesterol, diabetes, heart disease, and the like.

Domestic Patent No. 10-0717996 Domestic Patent No. 10-0375366 U.S. Patent 7,550,584

In order to solve the problems of the prior art as described above, the present invention is effective to extract rice hull and barley husk extract effective for the treatment and prevention of poultry, fish and livestock exposed to diseases caused by a number of pathogens such as viruses, bacteria, etc. It is an object to provide a pharmaceutical composition comprising as a component.

In addition, the present invention has an immune enhancing effect, can be used as a cholesterol lowering agent by reducing the amount of total cholesterol and LDL-cholesterol in the serum, showing a good effect on TNF-α derivatives can be used as a powerful anticancer drug It is an object to provide a composition.

It is another object of the present invention to provide a pharmaceutical composition that protects humans from infection caused by pathogens such as viruses, bacteria, parasites, and is effective in preventing and treating cancer.

It is another object of the present invention to provide a functional food composition effective for the prevention and treatment of viral diseases.

In order to achieve the above object, the present invention provides a pharmaceutical composition comprising a rice hull and barley husk extract as an active ingredient.

Specifically, the extract is preferably extracted from a mixture of rice husk and barley husk in a weight ratio of 1: 0.5 to 1: 5.

In addition, the extract comprises 0.01 to 99.9% by weight of the composition.

The extract is a step of drying after dipping a mixture of rice husk and barley husk in distilled water; Adding an aqueous NaOH solution to the dried raw material and heating it; Cooling the heated raw material to separate a solution dissolved in an NaOH aqueous solution; Adding and stirring HCl solution to the separated solution; Separating the precipitate produced by the stirring to obtain a crude extract; And vacuum drying the crude extract and freeze drying to obtain a water-soluble extract and an insoluble extract.

The pharmaceutical composition may further include rice, oats, bean husks, and the like.

The pharmaceutical composition exhibits antiviral, antibacterial, anticancer, immunopotentiating and cholesterol inhibitory activity, and particularly exhibits antiviral activity against poultry, fish and livestock.

In addition, the pharmaceutical composition shows activity against Newcastle disease, Gamboro, poultry, poultry cholera or Marek's disease, in particular against H1N1 (H1N1) virus and H5N1 (Birdflu) virus.

In another aspect, the present invention provides a functional food composition comprising a rice hull and barley husk extract as an active ingredient.

The composition of the present invention is effective in the treatment and prevention of poultry, fish and livestock exposed to diseases caused by a number of pathogens, such as viruses and bacteria, and protects humans from infections caused by pathogens such as viruses, bacteria and parasites. It can maintain a healthy state and has anticancer activity.

In addition, the composition of the present invention has an immune enhancing effect and can be used as a cholesterol lowering agent by reducing the amount of total cholesterol and LDL-cholesterol in the serum. In addition, the composition of the present invention shows an excellent effect on the TNF-α derivatives can be used as a powerful anticancer agent.

Figure 1 shows the FT-IR, H-NMR, C-NMR spectrum of the rice husk and barley husk extract obtained according to an embodiment of the present invention.
Figure 2 shows the TGA, UV, DSC spectrum of the rice husk and barley husk extract obtained according to an embodiment of the present invention.
Figure 3 shows the LC mass by LC-MS spectroscopy of water-soluble extracts and crude extracts extracted from rice hulls and barley husks according to one embodiment of the present invention.
Figure 4 shows the degree of recovery over time after administering the rice husk and barley husk bark extract obtained according to an embodiment of the present invention to chickens with Newcastle disease.
5 is a photograph of one day after six days after the administration of rice husk and barley husk extract obtained according to an embodiment of the present invention to chickens with Newcastle disease.
Figure 6 shows the degree of recovery over time after administering the rice husk and barley husk extract obtained according to an embodiment of the present invention to chickens with Gambo.
Figure 7 is a photograph of one day, 5 days after the administration of the rice husk and barley husk extract obtained according to an embodiment of the present invention to chickens with chicken poultry.
8 is a photograph of one day, five days after the administration of the rice husk and barley husk extract obtained according to an embodiment of the present invention to chickens with Marek's disease.
Figure 9 shows the H1N1 virus growth inhibitory activity of the rice husk and barley husk extract obtained according to an embodiment of the present invention.
FIG. 10 is a photograph of 7 days after 3 days of administration of rice hull and barley husk extracts obtained according to an embodiment of the present invention to the sharks infected with epidemic ulcer syndrome.
Figure 11 shows the activity against Vibrio bulpicus bacteria of the rice husk and barley husk extract obtained according to an embodiment of the present invention.
Figure 12 shows the degree of recovery over time after administering the rice husk and barley husk extract obtained according to an embodiment of the present invention to chickens infected with nasal cold.
Figure 13 shows the TNF-α inhibitory effect of the rice husk and barley husk extract obtained according to an embodiment of the present invention.
Figure 14 is to confirm whether or not infected by the rice husk and barley husk extract obtained according to an embodiment of the present invention to chicks, and then exposed to chickens infected with the disease.

Hereinafter, the present invention will be described in detail.

According to the present invention, there are rices which are not infected with viruses in some species of rice, and the present inventors include a component that is resistant to viruses or acts as an antiviral agent on rice husks. The present invention has been completed based on this idea, considering that the virus cannot be easily attacked even if it is not infected or infected by the virus.

Hereinafter, the rice and barley described in the present invention generally mean all kinds of rice and barley grown and harvested or grown wild. Specifically, even if the varieties and production regions of rice and barley are not specified, all Of course, it means all the species of rice and barley that can be cultivated and harvested in the wild, or obtained wild, and this does not limit the scope of the present invention.

In addition, the rice hull and barley husk extracts described below are all extracts, fractions and purified products obtained at each stage of extraction, fractionation and purification from rice hull and barley husk, diluents or concentrates thereof, dried products thereof, and the like. Of course it means all.

The pharmaceutical composition of the present invention comprises an extract extracted from the rice husk and barley husk as an active ingredient.

The rice may be used as a conventional rice, for example, Oryza sativa cv. Juanbyeo, Oryza sativa cv. ), Cultivated rice (Oryza sativa cv.Dongjinjubyeo-1ho) and wild rice Oriza collina (Oryza collina), Oryza alta, Oriza Ostrariensis ( Oryza, including Oryza australiensis, Oryza minuta, Oryza officinalis, Oryza brachyantha, and Oryza brathii Etc. can be used. In particular, the use of rice varieties without genetic modification is more desirable for obtaining an effective extract.

The barley may also be used as a conventional barley, for example ivory barley, baekrye barley, large barley, such as shell barley, white rice barley, white glutinous rice barley, rice barley, such as sap glutinous rice barley, Jinyang barley, barley barley, ascending barley, etc. Beer barley, nutritious barley, soft barley, green barley, such as friendly barley can be used.

After using the rice and barley as described above to obtain a husk according to a conventional method, the rice husk and barley husk is mixed in a proportion, and then the extract is extracted from this mixture.

The extract extracted from the rice hulls and barley husks may be extracted from a mixture of rice hulls and barley husks in a weight ratio of 1:05 to 1: 5, preferably 1: 1 to 1: 3. have. When the mixing ratio of the rice husk and barley husk is outside the above range, the efficacy of the main ingredients in the husk of the bark may be hindered by the barley ?? bark ingredient, and the efficacy is lost, Virus, antibacterial and anticancer effects are not obtained.

The extract of the rice husk and barley husk can be extracted by a conventional method. For example, about 1 to 15 times the volume, preferably about 5 to 10 times the amount of water, C1 to C4 lower alcohol such as ethanol, methanol or the like or a mixed solvent having a mixing ratio of about 1: 0.1 to 1:10 Preferably, water is added and cold extraction, hot water extraction, ultrasonic extraction, reflux cooling for about 0.5 hours to 2 days, preferably 1 hour to 1 day, at an extraction temperature of 20 to 100 ° C., preferably 50 to 100 ° C. Extraction, extraction of the shaker and the like is an extraction method, preferably extracted 1 to 12 times, preferably 3 to 4 times by extraction and then filtered by the extraction method of the shaker, the filtered extract is 20 to 100 ℃, preferably with a vacuum rotary concentrator. The residue was concentrated under reduced pressure at 40 ~ 70 ℃ dried by vacuum freeze dryer can be obtained by soluble in water, lower alcohol or a mixed solvent thereof.

Specifically, in the present invention, the rice husk and barley husk extract is a step of drying after dipping a mixture of rice husk and barley husk in distilled water; Adding an aqueous NaOH solution to the dried raw material and heating it; Cooling the heated raw material to separate a solution dissolved in an NaOH aqueous solution; Adding and stirring HCl solution to the separated solution; Separating the precipitate produced by the stirring to obtain a crude extract; And vacuum drying the crude extract and freeze drying to obtain a water-soluble extract and an insoluble extract.

More specifically, the rice husk and barley husk extract is soaked in 5L distilled water for 30 minutes on the basis of 1kg of raw material, using a pump (pump) to remove water, 15L of NaOH aqueous solution ( 0.2-1.0 N) is added and heated at reflux (80-95 ° C.) for 60-5 hours. After heating, it is cooled to 30-35 ° C. for 30-60 minutes, in which materials not dissolved in NaOH aqueous solution are precipitated at the bottom. Then, the dissolved solution is separated into a new reactor by pumping, and 1.5L HCL solution (1-2N) is quickly added to the separated solution until it reaches pH 1-2.5 and stirred for 5-10 seconds. At this time, precipitation occurs, which causes precipitation for about 30 minutes, and clear liquids of the upper layer are removed by pumping. And the solution containing the remaining precipitate (crude extract) is filtered through a 0.45㎛ membrane filter vacuum dried (about 90 ℃), and then freeze-dried (freeze drying) at 10 ~ 20 ℃ dissolved part (water-soluble extract) and precipitate (Insoluble extract) is obtained. At this time, in order to obtain the active ingredient in the form of precipitates, care should be taken not to exceed 3.5.

In the present invention has been described in the present invention a method for ideally obtaining an extract from the rice husk and barley husk, but the present invention is not limited to the method of separating the rice husk and barley husk extract. will be. Of course, the rice hull and barley husk extract included in the pharmaceutical composition of the present invention can be obtained separately by a method to be improved and developed in the past or in the future.

Rice husk and barley husk extract extracted by the above method has a molecular weight of about 300gram / mole to 40,000gram / mole, the extract contains a monosaccharide, such as beta glucan, polysaccharides and the like.

Rice husk and barley husk extract contained in the pharmaceutical composition of the present invention as described above exhibits antiviral activity, antibacterial activity, anticancer activity, immune enhancing and cholesterol inhibitory effect.

Specifically, the rice husk and barley husk extract shows antiviral activity and antibacterial activity against poultry, fish and livestock, Newcastle disease, Gaboro disease, chicken pox, poultry cholera, Marek's disease, epidemic ulcer syndrome, nasal cold Strong activity against and the like.

In particular, the rice husk and barley husk extract of the present invention exhibits very potent activity against H1N1 (H1N1) virus and H5N1 (Bird Flu) virus, and thus can be used as a very powerful therapeutic agent.

In the present invention, in order to confirm the antiviral, antibacterial, anticancer, immune enhancing and cholesterol inhibitory activity of the rice husk and barley husk bark extract, antiviral activity, antibacterial activity, anticancer activity in vitro and in vivo , Immunostimulatory properties and cholesterol removal properties were confirmed and assayed.

The pharmaceutical composition of the present invention comprises 0.01 to 99.9% by weight of the rice hull and barley husk extract based on the total weight of the composition. When the content of the extract in the composition is out of the above range, the desired degree of antiviral antibacterial and anticancer effects will not be obtained.

In addition, the pharmaceutical composition of the present invention may further include rice, oats, soybean hulls and the like in addition to the rice husk and barley husk extract.

The rice, oats, soybean hulls, etc. may be added in the form of an extract, pulverized powder, crushed granulated form, etc. according to a conventional method, of course, does not affect to obtain the desired effect in the present invention If not, the form is not limited.

The components are contained in 0.01 to 10% by weight relative to the rice hull and barley husk extract can further maximize the antiviral, antibacterial, anticancer activity effect of the present invention.

In addition, the pharmaceutical dosage forms of the rice hull and barley husk extract of the present invention may be used in the form of their pharmaceutically acceptable salts, and may also be used alone or in combination with other pharmaceutically active compounds, as well as in suitable collections. Of course it can.

The pharmaceutical composition comprising the rice husk and barley husk extract of the present invention comprises a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are those commonly used in the preparation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, silicic acid Calcium, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oils, and the like. It is not. The pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995), which can be used in pharmaceutically effective amounts. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to achieve the above-described efficacy or activity of the active ingredient of the present invention.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, mucosal administration, and eyedrop administration.

Suitable dosages of the pharmaceutical compositions of the present invention may be prescribed in various ways depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion, reaction sensitivity, etc. of the patient. Can be. Preferably, the dosage of the pharmaceutical composition of the present invention is 0.001-100 mg / kg body weight. In addition, when the pharmaceutical composition of the present invention is administered to poultry, fish, and livestock, it is administered in an amount of 0.01 to 0.4 g / head for poultry, 0.01 to 0.1 g / head for fish, and 0.01 to 0.4 g / head for livestock. It is good.

The pharmaceutical compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient, according to methods which can be easily carried out by those skilled in the art. Or by incorporating into a multi-dose container. The formulation may be in the form of solutions, suspensions, syrups or emulsions in oils or aqueous media, or in the form of extracts, powders, powders, granules, tablets or capsules, and may further comprise dispersants or stabilizers.

The present invention also provides a food composition (or functional food composition) comprising a rice hull and barley husk extract as an active ingredient.

The rice husk extract and barley husk extract can be added to food as it is safe for the human body can be applied as food for the prevention and treatment of viral diseases, bacterial diseases, cancer diseases and the like. Since the food composition of this invention obtained in this way can be ingested routinely, high antiviral, antibacterial, anticancer effect can be expected, and it is very useful.

The rice husk and barley husk extract is included in an amount of 0.01 to 90% by weight based on the total weight of the food composition. When the content of the extract in the composition is out of the above range, it can express the desired antiviral, antibacterial and anticancer effects to the maximum, and have high solubility in water to form a formulation with other components. In addition, it is an advantage that the drug can be administered through a route such as oral / intravenous use.

The food composition is not limited in kind, but, for example, by adding a conventional palate, such as ramen, raw noodles, noodles, tofu, cereals, bread, chewing gum, candy, confectionary, etc. It can be manufactured as a food and can also be applied as an edible pigment. In addition, it can be formulated into a common formulation such as tablets, granules, pills, hard capsules, soft capsules or liquid formulations, and can be prepared as a juice, a pouch, a drink, or the like. It goes without saying that components other than the above-mentioned components can be appropriately selected and blended by those skilled in the art according to the formulations.

In addition, the rice husk and barley husk extract included in the food composition of the present invention can effectively prevent the smell or flavor change of the food caused by oxidation, rancidity of fats and oils and discoloration of food. Therefore, the rice husk and barley husk extract of the present invention may be used for preserving these foods or maintaining the freshness and quality of foods for a long time by blending them with various conventional foods.

Such foods include not only typical foods but also beverages (including alcoholic beverages), fruit and processed foods thereof (e.g., canned fruits, jars, jam, maare marlade, etc.), fish, meat and processed foods Ham, sausage, corn oil, etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, Vegetable protein, retort food, frozen food, various seasonings (e.g., soybean paste, soy sauce, soy sauce, etc.), and the like.

When the composition of the present invention is made of a food composition, it includes not only the active ingredient of the present invention as described above, but also components commonly added in the preparation of food, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings, and the like. Contains flavors. Examples of the carbohydrates described above include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, oligosaccharides, etc.), polysaccharides (eg, dextrins, cyclodextrins). Or a conventional sugar such as xylitol, sorbitol, erythritol, or the like. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (rebaudioside A, glycyrzin), etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.

In addition to the above, the food 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 Salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components may be used independently or in combination.

Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.

Example 1. Preparation of Rice husk and Barley husk extract

The rice hulls and barley husks were mixed in a weight ratio (w / w) of 1: 2 and mixed properly by blending. 1 kg of the raw material was immersed in 5L distilled water for 30 minutes, and then water was removed using a pump. 15 L of 0.2N NaOH solution was added to the wet raw material and heated at 95 ° C. for 90 minutes, and then cooled at 30 to 35 ° C. for 30 to 60 minutes. At this time, the undissolved material is settled to the bottom, the dissolved material by pumping was separated into a new reactor. 1.5L of 2N HCL solution was quickly added to the separated solution (about 12L) until pH 1-2.5 was reached and stirred for 5-10 seconds. At this time, the precipitate is left to stand for 30 minutes to allow the precipitate to be generated, the clear liquid of the upper layer was removed by pumping. Then, the solution containing the remaining precipitate (about 3L, crude extract) was filtered through a 0.45㎛ membrane filter and vacuum dried (about 90 ℃), and then freeze dried at 10 ~ 20 ℃ (FREEZE DRYING) dissolved part ( Water soluble extract) and a precipitate (insoluble extract).

Example 2. Preparation of Rice husk and Barley husk extract

Except that in Example 1, the rice husk and barley husks were mixed in a weight ratio (w / w) of 1: 1.5, 0.3N NaOH solution was added, and 2N HCl solution was added at 1.0 L. It carried out similarly to Example 1.

Example 3. Preparation of Rice husk and Barley husk extract

Except that in Example 1, the rice husk and barley husks were mixed at a weight ratio (w / w) of 1: 1, 0.1N NaOH solution was added, and 0.3L of 2N HCl solution was added. It carried out similarly to Example 1.

Example 4. Preparation of Rice husk and Barley husk extract

Except that in Example 1, the rice husk and barley husks were mixed at a weight ratio (w / w) of 1: 3, 0.1N NaOH solution was added, and 0.3L 2N HCl solution was added. It carried out similarly to Example 1.

Example 5. Preparation of Rice husk and Barley husk extract

Except for using the rice husks and barley husks in Example 1, except that the rice husks, barley husks and rice powder mixed in a weight ratio (w / w) of 1: 2: 0.1 used in Example 1 It was carried out in the same manner.

Experimental Example 1. Characterization of the extract

The brown color extract extracted in Example 1 was characterized by FT-IR, H-NMR, C-NMR, TGA, UV, DSC, LC-MS spectroscopy to observe the properties and the results are shown in FIG. . In addition, possible structures of the extracts were identified and shown in Table 1 below.

Table 1 below is the data obtained from the analysis by LC-MS spectroscopy, the expected structural formula of Table 1 is the structure of the extract of Example 1.

Retention time (minutes) m / z [M + H] ⁺ Calculated Structural Formula MW Expected structural formula 2.63 165.0544 C ₉ H ₉ O ₃ 164 C ₉ H ₈ O ₃ 2.77 639.1909 C ₂₆ H ₂₇ O ₁₀ N ₁₀
C ₄₂ H ₂₇ O ₅ N ₂ 638 C ₂₆ H ₂₆ O ₁₀ N ₁₀
C ₄₂ H ₂₆ O ₅ N ₂ 3.28 361.0917 C ₁₈ H ₁₇ O ₈ 360 C ₁₈ H ₁₆ O ₈ 3.65 331.0810 C ₁₇ H ₁₅ O ₇ 330 C ₁₇ H ₁₄ O ₇ 3.72 527.1544 C ₄₀ H ₁₉ N ₂
C ₂₇ H ₂₇ O ₁₁ 526 C ₄₀ H ₁₈ N ₂
C ₂₇ H ₂₆ O ₁₁

Figure 1 shows (a) FT-IR, (b) H-NMR, (c) C-NMR spectra, Figure 2 shows (a) TGA, (b) UV, (c) DSC spectrum, Figure 3 shows the LC mass by LC-MS spectroscopy of (a) the water-soluble extract and (b) the crude extract.

The brown colored extract of Example 1 was found to have about 10% solubility in demineralized water / PBS and about 90% solubility in 0.2N NaOH solution. In addition, the UV absorbance peak of the extract was found at 272 nm.

Experimental Example 2 Antiviral Activity against Newcastle Disease

The rice husks and barley husks obtained in Example 1 were treated for 8 days at a dose of 0.5 g / kg / day for 5 chickens with Newcastle disease, and then 10.5 g / s for the next two days. The dose was increased in excess of kg / day.

As a result, as shown in Figure 4 all the chickens could be confirmed that the healthy recovery on the 10th day. 5 is a photograph of (a) chickens with Newcastle disease (day 1), (b) chickens treated with Newcastle disease after extract administration (day 6), the rice husk and barley husk of the present invention as shown in FIG. The extract was confirmed to have an excellent therapeutic effect against poultry with Newcastle disease.

Experimental Example 3. Antiviral Activity against Gamboro

Chickens were infected with Gambo, and each of the 10 chickens infected with Gambo was divided into two groups of five, and one group contained the rice husk and barley husk extracts obtained in Example 1 above. The dose was 0.4 g / chicken / 12 hours orally for 10 days, and the extract was not administered to the rest of the groups.

As shown in FIG. 6, administration of the rice husk and barley husk extract of the present invention was completely recovered, whereas the chickens of the group without the extract were all dead.

Through the above results, it was confirmed that the rice husk and barley husk extract of the present invention has an excellent therapeutic effect against poultry affected by Gamboro disease.

Experimental Example 4. Antiviral Activity against Chicken Pox

Five chickens with chicken poultry were treated with oral administration of the rice husk and barley husk crude extract obtained in Example 1 at 0.4 g / chicken / 12 h for 3 days, and the results were observed.

7 is a photograph of (a) chicken with chicken pox (Day 1), (b) chicken treated with chicken pox (Day 5) after administration of the extract, as shown in Figure 7 the rice husk and barley husk of the present invention The extract was confirmed to have an excellent therapeutic effect against poultry with chicken pox.

Experimental Example 5. Antiviral Activity against Marek's Disease

For 5 chickens with Marek's disease, the rice hulls and barley husks obtained in Example 1 were treated orally with 0.4 g / chicken / 12 hours for 3 days, and the results were observed. .

8 is a photograph of (a) chickens with Marek's disease (day 1), (b) chickens treated with Marek's disease after administration of the extract (day 5), as shown in Figure 8 the rice husk and barley of the present invention The husk extract was found to have an excellent therapeutic effect against poultry affected by Marek's disease.

Experimental Example 6. Activity against H1N1 (H1N1) virus

Two-fold dilution was performed to measure the titer of virus lines (A / Beijing / H1N1 / 95) distributed by the National Institutes of Health.

First, 5 μl of diluted virus strain (H1N1) was added to a 96 well V-type plate, and 50 μl of crude extract and water-soluble extract of rice husk and barley husk obtained in Example 1 were added. At this time, 50ul of PBS was put into the RBC control, and 50ul of 0.5% GP blood cells were added. And after shaking to mix well, it was left to stand at room temperature for one hour. Virus titers were then calculated to find 4 HA Unit virus.

HI test, after diluting the insoluble extract of the rice husk and barley husk obtained in Example 1 in two-fold step, and put into 25ul in V-type 96 well plate containing 5ul of viral strain (H1N1). Then, the mixture was allowed to stand at room temperature for 30 minutes, and 50 μl of 0.5% GP blood cells were put therein, followed by reading at room temperature.

As a result, as shown in Figure 9, it was confirmed that the crude extracts, water-soluble extracts and insoluble extracts of rice hulls and barley husks all have an activity to inhibit the growth of H1N1 virus, water-soluble extracts up to 16mg / ㎖, Crude extract up to 63 mg / ㎖, insoluble extract was confirmed to inhibit the H1N1 virus up to 0.25 mg / ㎖.

Experimental Example 7. Antiviral Activity against Pandemic Ulcer Syndrome

ANABAS TESTUDINEUS (Bangladesh: KOI) infected with Pandemic Ulcer Syndrome was treated by administering the rice husk and barley husk extract obtained in Example 1 at a dose of 0.05 g / day for 5 days. Was observed.

FIG. 10 is a photograph of a shark that is completely treated with epidemic ulcer syndrome at (a) a representative sample of idiosyncratic with ulcer syndrome, (b) 3 days after administration of extract, and (c) 7 days after administration of extract. As shown in Figure 10, the rice husk and barley husk extract of the present invention was confirmed that the treatment effect is excellent for the fish with epidemic ulcer syndrome.

Experimental Example 8. Antibacterial Properties on in vitro

Crude extracts of the rice hull and barley husk obtained in Example 1 on a RAW264.7 cell line having a Vibrio vulnificus bacterium (a mixture of a water-soluble component and an insoluble component, crude), Aqueous extract (water soluble) and insoluble extract (IS) were added at a concentration of 0.01%, respectively, to observe the degree of inhibition of NO production caused by Vibrio bullicus. At this time, PBS buffer solution was used as a control.

As shown in Figure 11, the rice husk and barley husk extract of the present invention was confirmed that completely inhibit the NO production, through the results of the present invention effectively inhibit the activity of the Vibrio bulpicus bacteria I could see.

Experimental Example 9. Antibacterial activity against nasal cold

48 chickens infected with nasal cold were divided into two groups of 24 dogs each, and one group contained the rice hull and barley husk extract obtained in Example 1 in an amount of 0.02 g / day, The dose was administered in an amount of 0.4 g / day for 3 days. As a result, as shown in FIG. 12, the chickens of the group administered with the extract at an amount of 0.4 g / day recovered, whereas the chickens of the group administered with the extract at an amount of 0.02 g / day were about 50%. The recovery rate was confirmed.

Through the above results, it was confirmed that the rice hull and barley husk extract of the present invention is excellent in the treatment effect against poultry disease caused by bacteria such as nasal cold.

Experimental Example 10. TNF-α Inhibitory Effect

In order to confirm the inhibitory effect on TNF-α, a tumor necrosis factor, of the rice husk and barley husk extracts obtained in Example 1, the method described in (Exp. Geront., 37, 235-247, 2002) In accordance with, the rice husk and barley husk extract obtained in Example 1 was evaluated for the ability to inhibit TNF-α production in rat peripheral blood monocytes (PBMC).

First, 6 ml of rat blood collected from the abdominal vein while being treated with ACD (acid / citrate / dextrose) was carefully added to a tube containing 6 ml of Ficoll 1077, and the tube was then subjected to 1500 rpm (415 G) at room temperature. Centrifuge for 30 minutes at. The cell fraction thus obtained (middle fraction) was suspended in a 50 ml tube containing phosphate buffered saline (PBS). The tube was centrifuged at 2,000 rpm (737 G) for 5 minutes. The resulting pellet was suspended in 6 mL PBS and 6 mL Picol 1077 and centrifuged at 1500 rpm (415 G) for 30 minutes at room temperature to remove residual plasma. The resulting cell fractions were suspended in PBS and centrifuged for 5 minutes at 2,000 rpm. This washing process was repeated twice. The recovered cells were then suspended in 10 ml RPMI-1640 containing 10% FBS and then diluted to contain 4 × 10 ⁶ cells per ml. Cell suspensions were dispensed at 100 μl per well of 96-well plates and pretreated with 1 μl of DMSO standard solution containing placebo or the indicated concentration of the compound of the invention. The plates were incubated for 1 hour at 37 ° C. under 5% CO ₂ and 95% O ₂ conditions and then RPMI-1640 100 containing 0.1 μg / ml LPS, 5U / ml IFN-γ and 10% FBS in each well. Μl was added. The plates were then incubated for 18 hours at 37 ° C. under 5% CO ₂ and 95% O ₂ conditions, and then the supernatants recovered from each well were analyzed by ELISA to measure TNF-α levels.

Mock of FIG. 13 is a control group without any component added, NBML-C50 is 50 μg / ml of crude extract, NBML-C100 is 100 μg / ml of crude extract, and LPS is lipopolysaccharide. It represents a substance that strongly promotes -α.

As a result, as shown in Figure 13, the rice hull and barley husk extract according to the present invention was confirmed that it can effectively lower the TNF-α level by inhibiting the production of TNF-α.

Experimental Example 11. Immune Enhancing Effect

Thirty day old chicks were born into three groups of 10 animals each, one group containing the rice hull and barley husk extract obtained in Example 1 in an amount of 0.02 g / day, the other group 0.4 It was administered for 8 days in the amount of g / day. No extract was administered to the rest of the group. Thereafter, all chicks were placed with the infected chickens to expose the disease.

As a result, as shown in Figure 14, the chicks of the group of rice hull and barley husk extract 0.4g / day all survived without infection with Gambo, whereas the chicks of the group administered at 0.02g / day 50% survival rate was confirmed. On the other hand, in the case of the group not administered the extract all died.

Through these results, it can be seen that the rice hull and barley husk extract of the present invention can protect poultry from viral and bacterial diseases with excellent immune effect.

Experimental Example 12 Cholesterol Inhibitory Activity

In order to measure the cholesterol inhibitory activity of the rice husk and barley husk extract obtained in Example 1, cholesterol and LDL-cholesterol levels in serum were measured in rats fed a high fat diet.

Positive Wister rats, about 80-100 g in weight, were supplied by the International Center for Diarrheal Disease Research in Bangladesh. In addition, the composition of the diet is beef (3 kg), starch (2.4 kg), sucrose (2.25 kg), flour (2 kg), hardened oil (1 kg), vitamin mix (10 kg), the rice pad of Example 1 Bark and barley husk extract (0%, 0.1%, 0.33%, 1% by weight) and carboxymethylcellulose (112.5 g total).

First, 40 positive Wister rats were separated into four groups of 10 rats, and then 0% by weight of the extract in the diet was added to the group 1, and 0.1, 0.33 and 1% by weight of the extracts were added to the groups 2 to 4, respectively. The diet was fed in 28 days ad libitum. It also did not limit the supply of water. After feeding for 28 days, the total cholesterol and LDL-cholesterol (triglyceride) levels in the serum of rats were measured, and the results are shown in Table 2 below.

The numerical values in Table 2 below are mean ± SEM values.

group Total cholesterol in serum LDL-cholesterol in serum One 86.4 ± 2.2 33.6 ± 1.9 2 67.9 ± 2.1 11.6 ± 1.3 3 64.3 ± 3.6 9.8 ± 1.9 4 57.9 ± 3.3 4.9 ± 0.7

As shown in Table 2, the cholesterol and LDL-cholesterol levels in the serum were lowered in the case of the 2-4 groups administered the rice husk and barley husk extracts according to the present invention compared to the one group without the extract. I could confirm it. In particular, as the concentration of the extract increased it was also found that the serum cholesterol and LDL-cholesterol inhibitory effect is also better.

Through these results, the rice husk and barley husk extract of the present invention was found to have excellent cholesterol inhibitory effect, it could be expected that it can be used as an excellent cholesterol inhibitor.

Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.

Claims (12)

A pharmaceutical composition for preventing and treating viral diseases, characterized in that it comprises rice husk and barley husk extract as an active ingredient. The method of claim 1,
The extract is a pharmaceutical composition for the prevention and treatment of viral diseases, characterized in that the extract from the mixture of rice husk and barley husk bark 1: 0.5 to 1: 5 mixed by weight ratio. The method of claim 1,
The extract is a pharmaceutical composition for preventing and treating viral diseases, characterized in that it comprises 0.1 to 99.9% by weight of the composition. The method of claim 1,
The extract is a step of drying after dipping a mixture of rice husk and barley husk in distilled water; Adding an aqueous NaOH solution to the dried raw material and heating it; Cooling the heated raw material to separate a solution dissolved in an NaOH aqueous solution; Adding and stirring HCl solution to the separated solution; Separating the precipitate produced by the stirring to obtain a crude extract; And vacuum drying the crude extract followed by freeze drying to obtain a water-soluble extract and an insoluble extract. The pharmaceutical composition for preventing and treating viral diseases, characterized in that it is extracted. The method of claim 1,
The composition is a pharmaceutical composition for preventing and treating viral diseases, characterized in that it further comprises any one or more components selected from rice, oats and soybean peel. The method of claim 1,
The composition is a pharmaceutical composition for preventing and treating viral diseases, characterized in that it exhibits antibacterial, anti-cancer, immune enhancing and cholesterol inhibitory activity. The method of claim 1,
The composition is a pharmaceutical composition for preventing and treating viral diseases, characterized in that it exhibits antiviral activity against poultry, fish or livestock The method of claim 1,
The composition is a pharmaceutical composition for preventing and treating viral diseases, characterized in that it exhibits activity against any one or more selected from Newcastle disease, Gamboro, chicken pox, poultry cholera, Marek's disease, pandemic ulcer syndrome and nasal cold. The method of claim 1,
The composition is a pharmaceutical composition for preventing and treating viral diseases, characterized in that it exhibits activity against H1N1 (H1N1) virus and H5N1 (Bird Flu) virus. Functional food composition for preventing and improving viral diseases, characterized in that the rice husk bark and barley bark extract as an active ingredient. 11. The method of claim 10,
The extract is a functional food composition for the prevention and improvement of viral diseases, characterized in that the extract from the mixture of rice hull and barley husks in a weight ratio of 1: 0.5 to 1: 5. 11. The method of claim 10,
The extract is a functional food composition for preventing and improving viral diseases, characterized in that it comprises 0.01 to 90% by weight of the composition.

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