KR101444098B1 - Novel use of rice, rice bran or rice hull extract as an histamine receptor antagonist - Google Patents

Abstract

The present invention provides a novel use as a histamine receptor antagonist of rice, rice bran or rice hull extract. Rice, rice bran or rice hull extract is used as a natural antihistamine, and can cause allergic rhinitis; Inflammatory bowel disease; asthma; bronchitis; throw up; Gastric and duodenal ulcers; Gastroesophageal reflux; Sleep disorders; unrest; And to prevent or treat depression, as well as to have substantially the same level or an improved effect of reducing the facial time and the sleeping time, as compared with the conventional diazepam, which is used as a sleep inducer. Rice, rice bran or rice husk extract is a natural product and has no side effect of forming cognitive impairment, tolerance or dependency even after prolonged use.

Description

A novel use of rice, rice bran or rice hull extract as a histamine receptor antagonist {Novel use of rice, rice bran or rice hull extract as an anti-histamine receptor antagonist}

The present invention relates to a novel use of a rice, rice bran or rice hull extract as a histamine receptor antagonist, and also to an allergic rhinitis containing rice, rice bran or rice hull extract as an active ingredient; Inflammatory bowel disease; asthma; bronchitis; throw up; Gastric and duodenal ulcers; Gastroesophageal reflux; Sleep disorders; unrest; And compositions for the prevention or treatment of depression.

Sleep constitutes one-third of human life and is the most fundamental and essential physiological phenomenon and is a very important factor in health maintenance and mental stability. Chronic sleep deprivation and disability negatively impacts overall physical and mental health, including cardiovascular disease and hypertension, memory and learning, metabolic control and weight, immunity and cancer resistance, diabetes, safety concerns, and mood. Currently, about 30% of the world's population is suffering from insomnia, and 10% is suffering from chronic sleep disorders.

According to a survey by the National Sleep Foundation USA, 2008, about 50% of US adults experience insomnia at least once a week, and about 50% of Americans are satisfied with their sleep . In recent years, the prevalence of insomnia and sleep disorders in Korea is rapidly approaching that of developed countries. The number of patients with sleep disorders has increased from 51,000 in 2001 to 228,000 in 2008, 4.5 times more than in the past eight years (National Health Insurance Corporation, 2009 ).

In Korea, there is not much interest in sleeping, and especially, there is no awareness of medical treatment or illness. On the other hand, according to statistics, about 15% of adults are known to need medication due to insomnia due to anxiety symptoms. Insomnia is caused by various factors such as stress, tension, and fear. Drugs include benzodiazepines Serotonin antagonists, etc. However, there is a serious problem in that cognitive impairment, tolerance, and dependence are formed when these drugs are used for a long period of time. In the United States, antihistamines and natural herbal products are used as non-prescription sleeping aids because of their low pharmacological side effects and dependence. In particular, antihistamines are antagonists to histamine receptors and are representative of cold medicines. It is the only sleeping pills that can be used.

Histamine [2- (4-imidazolyl) ethylamine] is one of the actinic neurotransmitters widely distributed throughout the body, for example, throughout the gastrointestinal tract [Burks 1994 in Johnson LR ed., Physiology of the Gastrointestinal Tract, Raven Press, NY, pp. 211-242]. Histamine has been implicated in gastric acid secretion, intestinal motility (Leurs et al., Br J. Pharmacol. 1991, 102, pp 179-185], vasomotor system response, intestinal inflammatory response and allergic response (Raithel et al., Int. Arch. Allergy Immunol. 1995, 108, 127-133) (Panula et al., Proc. Natl. Acad. Sci. USA 1984, 81, 2572-2576; Inagaki et al., J. Comp. Neurol 1988, 273, 283-300). To date, histamine mediates all of its actions in both the central nervous system and peripheral regions through the histamine receptors of the four families, the histamine H ₁ , H ₂ , H ₃ and H ₄ receptors. Histamine receptors and above in connection with the allergic and immune responses, gastric acid secretion, such as in the H _1, H _2, H ₃ and H ₄ receptor agonists alone or in combination, and allergic rhinitis, inflammatory bowel disease, asthma, bronchitis, vomiting Duodenal ulcer or gastroesophageal reflux, sedative in the brain and induction of sleep.

On the other hand, rice is a valuable food resource, one of the world's top three grains, which is half of the world's population, and is a key resource in the country that can not be confronted with other crops, especially in Asia. Rice, which is rice husk, which is the surface of rice, is called brown rice, rice rice with rice husk and rice husk is removed and rice is called white rice. Especially, rice bran is a very good material for nutrition though it is a by-product of Jeongbaekmi.

Rice bran (rice bran) contains about 95% of rice nutrients and contains high quality protein, dietary fiber and various vitamins and minerals. Antioxidants, antioxidants, anti-inflammatory, anti-arteriosclerosis, cholesterol-lowering, growth-promoting, digestive and immune-enhancing effects of rice bran have been found. However, the correlation between rice, rice bran or rice husk and histamine receptor antagonists, and furthermore, allergic diseases such as allergic rhinitis, asthma, diseases associated with excessive acid secretion, sleep, anxiety or depression have not been disclosed.

The present invention relates to histamine receptor antagonists, including allergic rhinitis; Inflammatory bowel disease; asthma; bronchitis; throw up; Gastric and duodenal ulcers; Gastroesophageal reflux; Sleep disorders; unrest; And to identify ingredients derived from natural materials that can replace conventional drugs that have been used in the prevention or treatment of depression.

In the present invention, " agonist ", when not otherwise indicated, interacts with histamine receptors, i.e., the H ₁ receptor, the H ₂ receptor, the H ₃ receptor and the H ₄ receptor to activate the histamine receptor and inhibit the physiological or pharmacological &Quot; antagonist " is a substance that competitively binds to a receptor at the same site as the agonist, does not activate the intracellular response initiated by the active form of the receptor, thereby inhibiting intracellular response by the agonist Of the material.

As histamine receptor antagonists, the present inventors have found that allergic rhinitis; Inflammatory bowel disease; asthma; bronchitis; throw up; Gastric and duodenal ulcers; Gastroesophageal reflux; Sleep disorders; unrest; And to produce a natural composition highly effective for the prevention or treatment of depression. As a result, the present invention has been completed by confirming that rice bran acts as a histamine receptor antagonist, and particularly has an effect of preventing or treating sleep disorder, anxiety or depression.

The present invention relates to a new use of rice, rice bran or rice hull extract as a histamine receptor antagonist, an allergic rhinitis comprising rice, rice bran or rice hull extract as an active ingredient; Inflammatory bowel disease; asthma; bronchitis; throw up; Gastric and duodenal ulcers; Gastroesophageal reflux; Sleep disorders; unrest; And a pharmaceutical composition for the prevention or treatment of depression, an allergic rhinitis comprising administering to a subject a therapeutically effective amount of rice bran extract or rice bran powder; Inflammatory bowel disease; asthma; bronchitis; throw up; Gastric and duodenal ulcers; Gastroesophageal reflux; Sleep disorders; unrest; And a method of preventing or treating depression.

In the present invention, improvement of sleep disorder, prevention or treatment may mean decrease in the time of the face, increase of the sleep duration, or increase of the sleeping surface.

In the following examples, rice, rice bran or rice hull extract exhibited almost the same level or an improved effect of reducing the facial time and the sleeping time as compared with the medicament diazepam, which is conventionally used as a sleeping agent, an anxiolytic or an antidepressant, . It is also known that the sleep effect of the pyrilamine maleate salt known as the histamine receptor antagonist is completely inhibited by 2-pyridylethylamine dihydrochloride, a histamine receptor agonist, , Rice bran or rice husk extract is suppressed and acts as a natural antihistamine agent. Therefore, rice, rice bran or rice husk extract is a histamine receptor antagonist, which is an allergic rhinitis; Inflammatory bowel disease; asthma; bronchitis; throw up; Gastric and duodenal ulcers; Gastroesophageal reflux; Sleep disorders; unrest; And compositions for the prevention or treatment of depression, and is particularly useful as a composition for preventing or treating sleep disorders, anxiety or depression. Unlike conventional sleeping pills, rice, rice bran or rice husk extract is harmless to humans, which is approved as food. It has no side effects, and has excellent induction effect on sleep and extension of sleeping. Therefore, it can prevent or treat sleep disorder, anxiety or depression Can be usefully used. In the examples of the present invention, an experiment for anxiety or depression was not performed separately. However, for example, ingredients used for improvement, prevention, or treatment of sleep disorders, such as diazepam, , Prophylactic or therapeutic, as is well known to those skilled in the art.

In the present invention, the rice, rice bran or rice hull extract may be extracted using rice bran water, an organic solvent, or a mixture thereof as an extraction solvent. The kind of the organic solvent used at this time and the mixing ratio of water and the organic solvent are not particularly limited.

For example, the organic solvent may be one or more solvents selected from the group consisting of lower alcohols, hexane, acetone, ethyl acetate, chloroform, and diethyl ether. The lower alcohol may be an alcohol having 1 to 6 carbon atoms. For example, as the lower alcohol, methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol can be used. The organic solvent may be a polar solvent such as acetic acid, dimethyl-formamide (DMFO) or dimethyl sulfoxide (DMSO), acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, 2,2,4-trimethylpentane, decane 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1- chloropropane, chlorobenzene , Non-polar solvents such as benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride and THF (tetrahydrofuran) may also be used.

As can be seen in the following examples, when rice, rice bran or rice husk is extracted with an organic solvent, the rice, rice bran or rice hull extract is separated into an upper layer liquid and a lower layer liquid. The upper liquid layer is an oil fraction of rice bran extract, Is a common solvent extract. When the lower layer is separated again through centrifugation or filter paper, it is divided into a liquid phase and a solid residue, which is a wax fraction of rice, rice bran or rice husk extract. Therefore, the rice, rice bran or rice husk extract of the present invention is interpreted as including all of the oil fraction of rice, rice bran or rice hull extract, the liquid fraction of rice bran extract, and the wax fraction of rice bran extract. In one embodiment, the rice, rice bran or rice husk extract may be at least one selected from the group consisting of rice, rice bran or rice husk extract oil fraction, rice bran extract fraction, liquid fraction of rice bran extract or chaff extract, and wax fraction of rice, . ≪ / RTI >

In one embodiment, the rice, rice bran or rice husk extract may be a lower alcohol extract of rice bran, preferably an ethanol extract of rice, rice bran or rice hull.

In one embodiment, the rice, rice bran or chaff extract may be a hexane extract of rice, rice bran or rice hull. The hexane extract of rice, rice bran or rice husk is generally used as a 'rice bran oil'. Thus, in one embodiment of the present invention, the rice bran extract may be uniquely rice bran. As used herein, the term " extract " also includes fractions obtained by further fractionating the extract. That is, the rice, rice bran or rice hull extract includes not only those obtained by using the above-mentioned extraction solvent, but also those obtained by further applying a purification process thereto. Further, fractions obtained by passing the above extract or fractions through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (manufactured for separation according to size, charge, hydrophobicity or affinity), and the like The fractions obtained through various purification methods are also included in the rice, rice bran or rice husk extract of the present invention.

In one embodiment, the rice, rice bran or rice hull extract may be a fraction obtained by fractionating an organic solvent extract of rice bran with a second organic solvent. Herein, the organic solvent extract of rice, rice bran or rice husk refers to an oil fraction of rice, rice bran or rice husk extract, a liquid fraction of rice, rice bran or rice husk extract, a wax fraction of rice, rice bran or rice hull extract And, in consultation, means liquid fraction of rice, rice bran or rice husk extract. Therefore, in one embodiment, the fraction obtained by fractionating the organic extract of rice, rice bran or rice husk with the second organic solvent may be a fraction obtained by fractionating the liquid fraction of the rice bran extract with the second organic solvent. In another embodiment, the rice, rice bran or rice husk extract may be a fraction obtained by fractionating a lower alcohol extract of rice, rice bran or rice husk with a second organic solvent. In another embodiment, the rice, rice bran or rice hull extract may be a fraction obtained by fractionating an ethanol extract of rice, rice bran or rice hull with hexane. As can be seen in the following examples, fractions of rice, rice bran or rice hull extract having a large amount of fat-soluble components of such rice, rice bran or rice hulls exhibit very good elevation and sleeping effect.

The term " extract " used herein to refer to rice, rice bran or rice husk includes not only crude extract obtained by treating the rice bran with an extraction solvent, but also processed products of rice, rice bran or rice hull extract. For example, rice, rice bran or rice husk extract may be prepared in powder form by further processes such as vacuum distillation and lyophilization or spray drying.

The rice, rice bran or rice hull extract of the present invention is meant to include rice, rice bran or rice husk processed products such as rice bran or rice husk powder formulated to be administered to animals in a broad sense, such as rice, rice bran or rice hull itself . Although the present invention has been conducted with rice, rice bran or rice husk extract, it is expected that those skilled in the art can achieve the desired effects in the form of rice, rice bran or rice husks.

In the present specification, the term "comprising as an active ingredient" means an amount sufficient to achieve the efficacy or activity of rice, rice bran or rice hull extract. In one embodiment of the present invention, the rice, rice bran or rice hull extract in the composition of the present invention may contain, for example, at least 0.001 mg / kg, preferably at least 0.1 mg / kg, more preferably at least 10 mg / More preferably at least 100 mg / kg, even more preferably at least 250 mg / kg, most preferably at least 0.1 g / kg. Since the rice, rice bran or rice husk extract is a natural product and there is no side effect on the human body even when administered in an excessive amount, the quantitative upper limit of the rice, rice bran or rice hull extract contained in the composition of the present invention can be selected by a person skilled in the art within a suitable range.

The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.

Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., .

The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient, Usually, a skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g / kg.

The pharmaceutical composition of the present invention may be formulated into a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Or by intrusion into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The present invention also provides a food composition for preventing or ameliorating sleep disorders, anxiety or depression comprising rice, rice bran or rice hull extract as an active ingredient.

The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectioneries, diet bars, dairy products, meat, chocolates, pizza, ram noodles, other noodles, gums, ice cream, .

The food composition of the present invention may contain, as an active ingredient, a rice bran extract or a rice bran powder, as well as a component that is ordinarily added during the manufacture of a food, for example, a protein, a carbohydrate, a fat, a nutrient, a seasoning agent, . Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared from a drink and a beverage, it additionally contains citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, various plant extracts and the like in addition to the rice, rice bran or rice hull extract of the present invention .

The present invention provides a health functional food comprising a food composition for preventing or ameliorating sleep disorder, anxiety or depression comprising the rice, rice bran or rice hull extract as an active ingredient. Healthy functional foods are foods made by adding rice bran extract or rice bran powder to food materials such as beverages, tea, spices, gum and confectionery, or by encapsulation, powdering, or suspension. However, unlike general medicine, there is an advantage that there is no side effect that can occur when a food is used as a raw material for a long period of taking the medicine. The health functional food of the present invention thus obtained is very useful because it can be ingested routinely. The amount of the rice bran extract or the rice bran powder added in such a health functional food can not be uniformly determined depending on the kind of the health functional food to be added, but may be added within a range that does not deteriorate the original taste of the food, Is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In the case of health functional foods in the form of pills, granules, tablets or capsules, they may be added usually in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

The present invention also provides the use of rice, rice bran or rice husk extract for the manufacture of medicaments or foods for the prevention, treatment or amelioration of sleep disorders, anxiety or depression. As described above, the rice bran extract or the rice bran powder can be used for prevention, treatment, or improvement of sleep disorder, anxiety or depression.

The invention also provides a method of preventing, treating or ameliorating sleep disorders, anxiety or depression, comprising administering to the mammal an effective amount of rice, rice bran or rice hull extract.

The term "mammal " as used herein refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.

As used herein, the term "effective amount" refers to the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in a tissue system, animal, or human, as contemplated by a researcher, veterinarian, physician or other clinician, ≪ / RTI > inducing a reduction of the symptoms of the disease or disorder. It will be apparent to those skilled in the art that the effective amount and the administration frequency of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the prevention, treatment, or improvement of the present invention, it is preferable to administer the rice, rice bran or rice husk extract at a dose of 0.001 mg / kg to 10 g / kg once or several times a day.

The composition comprising the rice, rice bran or rice hull extract as an active ingredient in the treatment method of the present invention can be administered orally or rectally, intravenously, intraarterally, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, And can be administered in a conventional manner.

Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described in detail below. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Is provided to fully convey the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

Rice, rice bran or rice hull extracts have been found to be effective as a medicament for diazepam, which is used as a conventional sleep inducer, The sleeping effect of rice, rice bran or rice husk extract is inhibited by the same mechanism that the sleeping effect of the maleate salt is completely inhibited by the histamine receptor agonist 2-pyridylethylamine dihydrochloride and acts as a natural antihistamine agent, It is derived from rice, rice bran or rice hull, and there is no side effect that cognitive impairment, resistance or dependency is formed even after long-term use.

Fig. 1 is a graph showing the effect of RWE on the surface time of a mouse administered with pentobarbital water surface dose (45 mg / kg, ip), Fig. 2 is a graph showing the effect of pentobarbital water surface dose (45 mg / FIG. 2 is a graph showing the effect of RWE on the sleeping time of the administered mice. FIG. After 45 minutes of oral administration (po), DZP (2 mg / kg) and RWE (50, 100, 250, 500 mg / kg) were administered to the control (0.5% CMC-saline 10 mL / kg) Each graph represents the mean value (means ± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP stands for diazepam.
FIG. 3 is a graph showing the effect of REE on the surface time of a mouse administered with pentobarbital water surface dose (45 mg / kg, ip), and FIG. 4 is a graph showing the effect of pentobarbital water surface dose (45 mg / FIG. 3 is a graph showing the effect of REE on the sleeping time of the mice administered. FIG. After 45 minutes of oral administration (po), DZP (2 mg / kg) and REE (50, 100, 250, 500 mg / kg) were administered to the control (0.5% CMC-saline 10 mL / kg) . Each graph represents the mean value (means ± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP represents diazepam.
FIG. 5 is a graph showing the effect of HWE on the surface time of a mouse administered with pentobarbital water surface dose (45 mg / kg, ip), and FIG. 6 is a graph showing the effect of pentobarbital water surface dose (45 mg / Lt; RTI ID = 0.0 &gt; HWE < / RTI &gt; After 45 minutes of oral administration (po) with DZP (2 mg / kg) and HWE (50, 100, 250, 500 mg / kg), pentobarbital was administered to the control (0.5% CMC-saline 10 mL / kg) Each graph represents the mean value (means ± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP stands for diazepam.
FIG. 7 is a graph showing the effect of HEE on the surface time of a mouse administered with pentobarbital water surface dose (45 mg / kg, ip), FIG. 8 is a graph showing the effect of pentobarbital water surface dose (45 mg / Lt; RTI ID = 0.0 &gt; HEE < / RTI &gt; to the sleeping time of the administered mice. After 45 minutes of oral administration (po) with DZP (2 mg / kg) and HEE (50, 100, 250, 500 mg / kg) in the control group (0.5% CMC-saline 10 mL / kg) . Each graph represents the mean value (means ± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP represents diazepam.
FIG. 9 is a graph showing the effect of BWE on the surface time of a mouse administered with pentobarbital water surface dose (45 mg / kg, ip), FIG. 10 is a graph showing the effect of pentobarbital water surface dose (45 mg / Lt; RTI ID = 0.0 &gt; BWE < / RTI &gt; to the sleeping time of the administered mice. After 45 minutes of oral administration (po) with DZP (2 mg / kg) and BWE (50, 100, 250, and 500 mg / kg), pentobarbital was administered in the control (0.5% CMC-saline 10 mL / kg) Each graph represents the mean value (means ± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP stands for diazepam.
FIG. 11 is a graph showing the effect of BEE on the time of face of a mouse to which pentobarbital water surface dose (45 mg / kg, ip) was administered, and FIG. 12 is a graph showing the effect of pentobarbital water surface dose (45 mg / Lt; RTI ID = 0.0 &gt; BEE < / RTI &gt; on the sleeping time of the administered mice. After 45 minutes of oral administration (po) with DZP (2 mg / kg) and BEE (50, 100, 250, 500 mg / kg) in the control group (0.5% CMC-saline 10 mL / kg) Each graph represents the mean value (means ± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP represents diazepam.
FIG. 13 is a graph showing the effect of BEE-Wax on the time of face of a mouse to which pentobarbital water surface capacity (45 mg / kg, ip) was administered, ) &Lt; / RTI &gt; on the sleeping time of mice. After 45 minutes of oral administration (po), DZP (2 mg / kg) and BEE-Wax (50, 100, 250, 500 mg / kg) were administered to the control (0.5% CMC-saline 10 mL / kg) . Each graph represents the mean value (means ± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP represents diazepam.
Fig. 15 is a graph showing the effect of BEE-Oil on the time of the face of a mouse to which pentobarbital water surface dose (45 mg / kg, ip) was administered, Fig. 16 is a graph showing the effect of pentobarbital water surface dose ) On the effect of BEE-Oil on the sleeping time of mice. After 45 minutes of oral administration (po), control (0.5% CMC-saline 10 mL / kg), DZP (2 mg / kg) and BEE-Oil (50, 100, 250, 500 mg / kg) Each graph represents the mean value (± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP represents diazepam.
17 is a graph showing the effect of BEE (500 mg / kg) on elevation times and total sleep time in SD rats. Each graph represents the mean value (± SEM, n = 8). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON represents a control group.
Figure 18 shows the effect of BEE (500 mg / kg) on sleep structure in SD rats. CON represents a control group.
19 is a graph showing changes in awakening, non-REM level and REM level according to time. CON represents a control group.
20 is a view showing the fractionation process of BEE (EtOH extract). EtOAc is ethyl acetate; n-BuOH is n-butanol; H2O represents distilled water.
FIG. 21 is a graph showing the effect of BEE fraction on the surface time of a mouse administered with pentobarbital water surface dose (45 mg / kg, ip), FIG. 22 is a graph showing the effect of pentobarbital water surface dose (45 mg / Lt; RTI ID = 0.0 &gt; BEE < / RTI &gt; (Control) (0.5% CMC-saline 10 mL / kg), DZP (2 mg / kg) BEE-H, BEE-B, BEE-W and BEE- Forty-five minutes later, pentobarbital was administered. Each graph represents the mean value (± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DZP is diazepam; H is hexane extract; B is butanol extract; W is water extract; E represents ethyl acetate extract.
23 is a view showing a process of separating a sub-fraction of n-hexane extract from BEE. EtOAc is ethyl acetate; H2O represents distilled water.
24 is a graph showing the effect of the sub-fraction obtained from BEE-H on the time of face of a mouse to which pentobarbital water surface capacity (45 mg / kg, ip) was administered and FIG. 25 is a graph showing the effect of pentobarbital water surface capacity / kg, &lt; / RTI &gt; ip) on the sleeping time of mice. Pentavaval was administered 45 minutes after oral administration (po) of control (0.5% CMC-saline 10 mL / kg), DZP (2 mg / kg) and BEE-H subfraction (50 mg / kg). Each graph represents the mean value (± SEM, n = 10). * Means p <0.05 compared with the control, ** means that there is a significant difference (Dunnet's test) at p <0.01 compared with the control group. C is the control; D represents diazepam.
FIG. 26 is a graph showing the effect of sub-fraction obtained from BEE-H on the time of face of a mouse to which pentobarbital water surface capacity (45 mg / kg, ip) was administered and FIG. 27 is a graph showing the effect of pentobarbital water surface capacity / kg, &lt; / RTI &gt; ip) on the sleeping time of mice. Pentavaval was administered 45 minutes after oral administration (po) of control (0.5% CMC-saline 10 mL / kg), DZP (2 mg / kg) and BEE-H subfraction (250 mg / kg). Each graph represents the mean value (± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. C is the control; D represents diazepam.
28 is a view showing a process of separating a small fraction of n-hexane extract from BEE. MeOH is methanol; EtOAc represents ethyl acetate.
FIG. 29 is a view showing a process of separating a small fraction of n-hexane extract. FIG. EtOAc is ethyl acetate; CHCl3 is chloroform; MeOH is methanol; H2O represents distilled water.
30 is a view showing a process of separating a small fraction of n-hexane extract. EtOAc is ethyl acetate; CHCl3 is chloroform; MeOH represents methanol.
FIG. 31 is a graph showing the effect of gamma oryzanol on the sleeping time of mice administered with pentobarbital water surface dose (45 mg / kg, ip), FIG. 32 is ferulic acid, FIG. 33 is octacosanol, , Fig. 35 is a graph showing the effect of beta-sitosterol, Fig. 36 is linoleic acid, and Fig. 37 is the effect of oleic acid. (0, 25, 50, 100 mg / kg), gamma oryzanol, ferulic acid, octacosanol, stigmasterol, beta-sitosterol, linoleic acid or oleic acid (30 mg / kg) Was administered orally (po) and pentobarbital was administered after 45 minutes. Each graph represents the mean value (means ± SEM, n = 10). * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control. CON is the control; DH represents dorsepepin hydrochloride.
FIG. 38 is a graph showing the effect of the BEE-H-2 fraction on the activity of GPCR (G-Protein Coupled Receptor).
FIG. 39 shows the time taken for the administration of the histamine receptor agonist (PD) to the rice water extract (RWE) of Production Example 1 and the antagonist (PMS) acting on the histamine receptor. FIG. (PDE) administration to extracts (RWE) and antagonists (PMS) acting on histamine receptors. Oral doses were given 45 minutes before pentobarbital administration (45 mg / kg) with control (CON, 0.5% CMC-saline 10 mL / kg), RWE (500 mg / kg) and PMS 70 mg / kg. * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control.
FIG. 41 shows the time taken for the administration of the histamine receptor agonist (PD) to the rice bran ethanol extract (BEE) of Production Example 3 and the antagonist (PMS) acting on the histamine receptor, and FIG. 42 shows the appearance time of rice bran ethanol (PD) administration to an extract (BEE) and an antagonist (PMS) acting on histamine receptors. Oral doses were given 45 minutes before pentobarbital administration (45 mg / kg) with control (CON, 0.5% CMC-saline 10 mL / kg), BEE 500 mg / kg and PMS 70 mg / kg. * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control.
FIG. 43 is a graph showing the onset time of diazepam (DZP) and PMS according to histamine receptor agonist (PD) administration, and FIG. 44 is a graph showing the time of onset of diazepam (DZP) and PMS according to histamine receptor agonist FIG. Oral doses were given 45 minutes before Pentavaval administration (45 mg / kg) with control (CON, 0.5% CMC-saline 10 mL / kg), DZP (2 mg / kg) PD (20 mg / kg) was intraperitoneally injected 10 minutes before PMS oral administration. * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control.
FIG. 45 is a graph showing the onset time of diazepam (DZP) and PMS according to the administration of the GABAA-benzodiazepine antagonist flumazenil (FLU), and FIG. 46 is a graph showing DZP ) And sleep time of PMS. Oral administration of pentobarbital (hypnotic dosage 45 mg / kg) 45 minutes before the control (CON, 0.5% CMC-saline 10 mL / kg), DZP (2 mg / kg) FLU (8 mg / kg) was intraperitoneally injected 10 minutes before oral administration of DZP or BEE. * Indicates p <0.05 compared to the control, ** indicates that there is a significant difference (Dunnet's test) at p <0.01 compared to the control.

[ Example ]

[Experimental Method]

Experimental animal

ICR mice (18-22 g, male) and SD rats (200-250 g, male) were distributed from Koatech and Orient Bio Co., respectively, and were adapted for one week in an experimental animal breeding box. Respectively. Animals were fed at a temperature of 23 ± 1 ℃, a humidity of 55 ± 5%, a light - dark cycle (9:00 am - 9:00 pm) and an illuminance of 3000 Lux. All animals were maintained in accordance with the Guidelines for the Use of Laboratory Animals by the Korea Food Research Institute (KFRI-IACUC, Korea Food Research Institutional Animal Care and Use Committee).

Pentobaby  Sleep induction experiment Pentobarbital - induced sleep test )

The pentobarbital sleep induction experiment was conducted within a certain time from 1 pm to 5 pm, and 10 mice (n = 10) per group were used after fasting for 24 hours before the experiment. All samples were prepared using 0.5% CMC-saline solution and administered orally (po) to mice 45 minutes before pentobarbital administration. In the general control group, 0.5% CMC-saline solution was treated at a concentration of 10 mg / kg, and diazepam was used as a positive control to compare the effect of the extract on sleep. Pentovavital (Hanlim Pharmaceutical Co., Ltd.) was administered via intraperitoneal injection (ip) at a concentration of 45 mg / kg (hypnotic dosage) according to the experimental design. After the pentobarbital treatment, each individual was moved to a separate space to measure sleep latency and sleep duration. The time of the elevation was set as the elapsed time until the righting reflex was lost for more than 1 minute after the peritoneal injection of pentobarbital, and the sleeping time was set as the time until the regular reflection was restored again. Mice that did not show sleep behavior after 10 minutes of pentobarbital administration were excluded from the experiment.

Sleep structure analysis

Electroencephalogram (EEG) and Electromyogram (EMG) measurements were performed after an adaptation of SD rats (Sprague-Dawley Rat, 200-250 g) for 1 week. The rats were anesthetized with pentobarbital (50 mg / kg, ip) and the head was fixed in a stereotaxic instrument. After cutting the subcutaneous connective tissue, a stainless-steel screw and a silver electrode line were inserted to measure EEG and EMG. After that, it was fixed with dental cement and sutured. Surgical site disinfection and antibiotic treatment were performed for 3 days to prevent the inflammation caused by the operation, and the recovery period was 7 days. In order to adapt to the measurement environment, the recording medium was connected with the 0.5% CMC-saline solution (po) used for the control group for 4 days prior to the measurement and was induced to conform to the experimental procedure. Electroencephalography (EEG) and electromyogram (EMG) were performed for 6 hours from 10:00 to 16:00 using PAL-8200 series (Pinnacle Technology Inc, Oregon, USA) after stabilization for 5 minutes after oral administration. ] Were measured. The sampling rate of the EEG and EMG was set to 200 Hz (epoch time: 10 s), the EEG was set to 0.1 to 25 Hz, and the EMG was set to a filter area of 10 to 100 Hz. Sleep structure analysis was performed by FFT (fast Fourier transform) algorithm and SleepSign program (Ver. 3.0, Kissei Comtec, Nagono, Japan) was used. The results of the analysis were divided into wake, REM sleep, rapid eye movement, theta band (6-10 Hz), NREM sleep, non-rapid eye movement, and delta band (0.65-4 Hz) . The elevation time was set as the time taken for a continuous 10 seconds epoch unit to appear more than 12 times.

Manufacturing example  1: Preparation of rice extract

Rice water extract (RWE) was prepared by adding 1 L of distilled water to 100 g of the ground sample and heating it to 100 ℃ for 1 hour. The rice ethanol extract (REE) was added to the ground sample 10 times (w / v) 50 ℃ incubator for 1 day and then ultrasonicated three times for 90 minutes per day. All the extracts were filtered and concentrated under reduced pressure, lyophilized and powdered for use in the experiment.

The yield of water extract (RWE) of rice was 1.16% by weight and the yield of ethanol extract (REE) of rice was 0.19% by weight.

Manufacturing example  2: Preparation of rice hull extract

(HWE) was prepared by adding 1 L of distilled water to 100 g of ground sample and heating it to 100 ℃ for 1 hour. The rice husk ethanol extract (HEE) was prepared by adding 10% (w / v) 50 ℃ incubator for 1 day and then ultrasonicated three times for 90 minutes per day. All the extracts were filtered and concentrated under reduced pressure, lyophilized and powdered for use in the experiment.

The yield of water extract (HWE) of rice hull was 5% and the yield of ethanol extract (HEE) of rice hull was 0.96%.

Manufacturing example  3: Preparation of rice bran extract

Brewer's water extract (BWE) was prepared by adding 1 L of distilled water to 100 g of ground sample and heating it at 100 ° C for 1 hour. The rice bran ethanol extract (BEE) was prepared by adding 10% (w / v) And extracted for 1 day in a 50 &lt; 0 &gt; C incubator. All the extracts were filtered and concentrated under reduced pressure, lyophilized and powdered for use in the experiment. The yield of water extract (BWE) of rice bran was 5.56% and the yield of ethanol extract (BEE) of rice bran was 7.02%.

BEE was identified as BEE-Wax and BEE-Oil in addition to BEE in addition to BEE. Specifically, the rice bran ethanol extract (BEE) was divided into three types during the manufacturing process. The first rice bran extract is divided into the supernatant and the lower layer. The supernatant is the oil fraction (BEE-Oil) of the rice bran extract, and the lower layer corresponds to the normal ethanol extract. When the lower layer is separated again through centrifugation or filter paper, it is divided into liquid phase (liquid fraction of rice bran extract) and solid residue, which corresponds to the wax fraction (BEE-Wax) of rice bran extract.

Experimental Example  1: Using rice extract Pentobaby  Sleep induction experiment

Water (RWE) and ethanol extract (REE) of rice of Preparation Example 1 were orally administered (po) at a concentration of 50, 100, 250 and 500 mg / kg and then treated with pentobarbital (45 mg / kg, ip) Induced sleep induction. As a result, as shown in FIG. 1 and FIG. 2, in the water extract of rice (RWE), the dose-dependent decrease in the face time and the increase in the sleep duration ( p <0.01) appear. Also, as can be seen in FIGS. 3 and 4, the ethanol extract (REE) of rice showed a dose-dependent decrease in the facial time and an increase in the sleep duration at concentrations other than 50 mg / kg. (REE) of rice showed higher increase of sleep duration than that of water extract of rice (RWE), while that of water extract (RWE) of rice and ethanol extract of rice (REE) Respectively.

Experimental Example  2: Using rice husk extract Pentobaby  Sleep induction experiment

(HWE) and ethanol extract (HEE) of Preparation Example 2 were orally administered (po) at a concentration of 50, 100, 250 and 500 mg / kg and then treated with pentobarbital (45 mg / kg, ip) Induced sleep induction. As a result, as can be seen from FIGS. 5 and 6, the water uptake time of the rice hull extract (HWE) was significantly decreased, and at the concentrations of 250 and 500 mg / kg, ( P <0.01), respectively. In addition, as can be seen from Figs. 7 and 8, the ethanol extract (HEE) of rice hull showed significant decrease in the face time ( p < 0.01) at concentrations except 50 mg / kg, Sleep duration was increased ( p <0.01). (HEE) of rice hulls showed a significant increase in the duration of sleep, while that of the rice hull extract (HWE) and rice husk ethanol extract (HEE) Respectively.

Experimental Example  3: Using rice bran extract Pentobaby  Sleep induction experiment

(BWE) and ethanol extract (BEE) of rice bran (BE) of Preparation Example 3 were orally administered (po) at a concentration of 50, 100, 250 and 500 mg / kg, followed by pentobarbital ip) were analyzed. As a result, it was found that all of the time of the elevation in the BWE was significantly decreased (p <0.01) (Fig. 9), and the elevation time decreased compared with the positive control DZP. In addition, an increase in dose-dependent sleep time was seen at concentrations of 250 mg / kg and 500 mg / kg except for 50 mg / kg and 100 mg / kg in BWE (Fig. 10). In BEE, as in BWE, there was an increase in the dose-dependent sleep time at concentrations of 250 mg / kg and 500 mg / kg, as well as reduced face time (Fig. 11) compared to DZP (Fig. 12). Sleep inducer BE showed significantly reduced elevation times and significantly increased sleep times at 250 mg / kg and 500 mg / kg without difference in water and ethanol extracts.

BEE-Wax was administered at a concentration of 50, 100, 250 and 500 mg / kg, BEE-Oil at a concentration of 500 and 1,000 mg / kg, and oral administration of BEE- After the administration (po), pentobarbital (45 mg / kg, ip) was intraperitoneally injected to measure the changes of the time of surface and sleeping time. Experimental results show that BEE-Wax has remarkable results. (P < 0.01) (Fig. 13) and increased sleep time ( p < 0.01) (Fig. 14), showing decreased incidence time and increased sleep time than BEE. ( P <0.01) (Fig. 15) and increase in sleeping time ( p <0.01) (Fig. 16) even at the concentration of BEE-oil 1,000 mg / kg. Although BEE and BEE-Wax did not induce much sleep inducing effect, BEE-Oil was able to confirm sleep enhancement effect, and BEE-Wax is expected to have a very large ripple effect in the future.

Experimental Example  4: Analysis of sleep structure using rice bran extract

Figure 17 shows the onset time and total sleep time after oral administration of BEE 500 mg / kg. The mean elevation time of the control group was 31.9 minutes and the BEE elevation time was 26.2 minutes, which was shortened by about 5.7 minutes. In the total sleep time, the control group was 165.4 minutes, and when BEE 500 mg / kg was orally administered, it increased by about 83 minutes to 248.6 minutes.

The awakening, RAM and non-time of SD rats were analyzed and shown in Fig. In the BEE-treated group, the increase in the number of nonremes decreased relative awakening, the control group had a normalization of 153.8 minutes, BEE 500 mg / kg of nonem increased 234.8 minutes by 81 minutes, and most of the increase in total sleep time .

FIG. 19 shows the ratio of awakening, non-remem, and rem in each hour while measuring brain EEG after oral administration of BEE 500 mg / kg. After 3 hours of oral administration, the nonremem increased significantly, then decreased slightly and then increased again. This led to sleeping in the first 3 hours of digestion of BEE, and overall the rate of nonem at all times was greater than in the control group, indicating that the sleep effect persisted for 6 hours. BEE did not significantly reduce sleep induction time, but it was judged to increase sleep persistence with longer total sleep time.

Manufacturing example  4: Preparation of solvent fraction of rice bran ethanol extract

BEE (liquid fraction of the rice bran ethanol extract of Preparation Example 3) was separated and purified by using four kinds of solvents such as hexane, butanol, water and ethyl acetate in order to obtain a more accurate single sleeping inducer by using the BEE having a high fat content 20).

200 kg of rice bran ethanol extract (BEE) was extracted at 60 ° C in a 60% aqueous EtOH solution for 43 hours. The extract was filtered and the residue was extracted three more times in the same manner. The resulting filtrate was combined and concentrated under reduced pressure to obtain 20 L of an aqueous EtOH extract. To the obtained EtOH extract, 7 L of H ₂ O was added and the mixture was extracted with n-hexane (27 L × 2) / H ₂ O (27 L) The H ₂ O fraction was extracted again with EtOAc (27 L × 2), and the H ₂ O fraction was again extracted with n-BuOH (25 L × 2). Concentration under reduced pressure afforded the respective layers, n- hexane fraction (BEE-H, 2176 g) , EtOAc fraction (BEE-E, 1458 g) and n-BuOH fraction (BEE-B, 1223 g) and H ₂ O fraction (BEE- W, 6.74 kg) was obtained (Figure 20)

Experimental Example  5: Solvent fraction of rice bran ethanol extract Pentobaby  Sleep induction experiment

In order to examine the effects of BEE-H (hexane), BEE-B (butanol), BEE-W (water) and BEE-E (ethyl acetate) on sleep induction, 50 mg / kg and 250 mg / After the administration (po), pentobarbital (45 mg / kg, ip) was intraperitoneally injected to examine changes in the time of the surface and sleeping time (FIGS. 21 and 22).

As a result of the experiment, BEE-E (ethyl acetate) showed a significant decrease in the time of the face, but no increase in the sleeping time was found, and it was confirmed that there was no improvement in the sleep induction. BEE-H (hexane) and BEE-B (butanol) showed a significant decrease in the surface time ( p <0.01) and an increase in the sleeping time ( p <0.01) ). ( P <0.01) and concentration-dependent sleeping time ( p <0.01) in BEE-H (hexane) and BEE-B (butanol) BEE-H (hexane) was found to be effective in promoting sleep induction. Finally, BEE-H (hexane) was used to proceed with more detailed separation and purification.

Manufacturing example  5: Rice bran extract of ethanol Hexane  Preparation of sub-fraction of fraction

In the solvent fraction of rice bran extract ethanol (BEE) In vivo experiments, a sub-fraction of the n-hexane fraction was prepared because the activity of the n-hexane fraction was significantly better than the other fractions. 365 g of n-hexane fraction (BEE-H, 2176 g) was subjected to SiO ₂ column chromatography (hereinafter referred to as cc), in which the column was packed with a silica gel resin having a diameter of 13 cm and a height of 15 cm. The elution solvent was gradually polarized from a non-polar solvent of n-hexane: EtOAc = 10: 1 and used at a ratio of n-hexane: EtOAc = 7: 1 2: 1. As a result, a total of 12 sub- To BEE-H-12) (see Fig. 23).

Experimental Example  6: BEE For the sub-fraction of -H fraction Pentobaby  Sleep induction experiment

To confirm the effect of BEE-H-1 to BEE-H-11 sub-fractions of the BEE-H fraction on the induction of sleep induction, oral administration (po) was carried out at a concentration of 50 mg / kg and 250 mg / kg, (45 mg / kg, ip) was injected intraperitoneally.

As shown in FIG. 24 and FIG. 25, at the concentration of 50 mg / kg, the increase in the sleeping time ( p <0.01) was significant only in the BEE-hexane extract 2 fraction, ( P <0.05) in extracts 2, 6, 10 and 11 fractions. As shown in FIG. 26 and FIG. 27, at the concentration of 250 mg / kg, significant increase in sleeping time ( p <0.01) was observed in the fractions of BEE-hexane extracts 1, 2, 4, 7, 10 and 11 BEE-hexane extract. ( P <0.05, p <0.01) in all fractions except 9 and 10 fractions. In the 11 fractions, it was found that the extracts of BEE-hexane extracts 2, 10 and 11 had a prolonged sleep inducing effect.

Manufacturing example  6: BEE From the sub-fraction of -H Fractional  Produce

(One) BEE -H-2 fraction

A total of 12 fractions (BEE-H-1 to BEE-H-12) were obtained by fractionation of BEE-H. About these in The activity of BEE-H-2, BEE-H-10 and BEE-H-11 fraction was higher than that of BEE-H-2. HEE-2 (8.2 g) was subjected to ODS cc (φ 4 × 7 cm, acetone: H ₂ O = 1: 1), and the same fraction was collected again to give five fractions (BEE- 1 to BEE-H-2-5), and substances were confirmed by SiO ₂ and ODS TLC (FIG. 28).

(2) BEE -H-10 fraction

A total of 12 fractions (BEE-H-1 to BEE-H-12) were obtained by fractionation of BEE-H. About these in The activity of the BEE-H-10 and BEE-H-11 fractions showed high activity in the BEE-H-2, BEE-H-10 and BEE-H-11 fractions. About BEE-H-10 (9.5 g ) SiO 2 cc (φ 6 × 17 cm, n-hexane: EtOAc = 10: 1 6: 1 4: 1 2: 1 1: 1 CHCl 3: MeOH = 8: 1 (BEE-H-10-1 to BEE-H10-23), and the material was identified by SiO ₂ and ODS TLC (see FIG. 29).

(3) BEE -H-11 fraction

A total of 12 fractions (BEE-H-1 to BEE-H-12) were obtained by fractionation of BEE-H. The activity of BEE-H-2, BEE-H-10 and BEE-H-11 fraction was higher than that of BEE-H-11. Respectively. About BEE-H-11 (810 mg ) SiO 2 cc (φ 5 × 10 cm, n-hexane: EtOAc = 10: 1 5: 1 3: 1 1: 1 CHCl 3 - MeOH = 8: 1 5: 1 (BEE-H-11-1 to BEE-H11-13), and the substance was identified by SiO2 and ODS TLC (see FIG. 30).

Experimental Example  7: Rice bran extract and BEE Analysis of surface components of -H-11 fraction

(BEE-E) of the rice bran ethanol extract of Preparation Example 4, the BEE-H-11 fraction of Preparation Example 5 and the rice bran ethanol extract (BEE) of Preparation Example 3, The contents of gamma oryzanol, myoinositol, ferulic acid, octacosanol, stigmasterol, beta - sitosterol and linoleic acid and oleic acid in fatty acids, which are known as useful components of rice or rice bran, were analyzed.

For the determination of gamma oryzanol, a standard sample was prepared for each concentration of oryzanol mixture using methylene chloride. The concentration of each oryzanol mixture and sample was adjusted to 20,000 ppm by using an HPLC column, Agilent TC-C18 (2) 4.6 x Acetonitrile and methanol were used as the mobile phase and acetonitrile and methanol used as the mobile phase for HPLC analysis were purchased from JT Baker (J.-Y. Cho et al. / Journal of Cereal Science 55 (2012) 337-343].

The standard samples required for GCMS were prepared at each concentration, and the standard samples and each sample were prepared at a concentration of 20,000 ppm. For the GC, the concentrations of the myoinositol, ferulic acid, octacosanol, stigmasterol, betasitosterol, linoleic acid and oleic acid Solvent (CHCl ₃ , pyridine, methanol) was used, and BSTFA for TMS was purchased from Sigma Aldrich and measured using agilent 6890 GC.

division The control (BEE-E) BEE
BEE-H-11 Gamma oryzanol (mg / g) 0 9.160 11.231 Myoinositol (占 퐂 / g) 0.518 0.616 0 Ferric acid ([mu] g / g) 0.023 0.403 0.222 Octacosanol (/ / g) 0 0.257 0.225 Stigmasterol (/ / g) 0 0.820 0.063 Beta-sitosterol ([mu] g / g) 0 3.393 0.147 Linoleic acid (占 퐂 / g) 0 0.173 1.481 Oleic acid (占 퐂 / g) 0 0.213 3.341

Gamma orzanol, ferulic acid, octacosanol, stigmasterol, beta-sitosterol, linoleic acid and oleic acid were commonly detected in extract (BEE) and fraction (BEE-H-11) exhibiting sleeping activity. In particular, it was confirmed that the contents of gamma oruzanol, ferulic acid and octacosanol were clearly distinguished from the control (BEE-E).

Therefore, the extract (BEE) or fraction (BEE-H-11) which exerts a sleeping activity is composed of 5 to 20 mg / g of gamma oryzanol, 0.1 to 1 μg / g of ferulic acid, 0.1 to 1 占 퐂 / g.

Experimental Example  8: Rice bran extract and BEE -H-11 fraction Of the resulting component Pentobaby  Sleep induction experiment

(BEE) and fractions (BEE) contained in the extract (BEE) and the fraction (BEE-H-11) of Experimental Example 7, which are Gamma orolzanol, Ferulic acid, Octacosanol, Stigmasterol, Beta sitosterol, Linoleic acid, Ferulic acid, octacosanol, stigmasterol, beta-sitosterol, linoleic acid, and oleic acid at concentrations of 10, 25, 50, and 100 mg / kg, respectively, (45 mg / kg, ip) after oral administration (po).

31 to 37, sleeping time tended to increase as the content of gamma oruzanol, ferulic acid, octacosanol, stigmasterol, beta-sitosterol, linoleic acid and oleic acid increased. However, in order to show a significant increase in sleeping time, It was confirmed that administration of 25 to 50 mg / kg was necessary.

Therefore, considering the content of single compounds such as gamma oryzanol, ferulic acid, octacosanol, stigmasterol, beta-sitosterol, linoleic acid and oleic acid contained in the extract (BEE) and the fraction (BEE- None of these single compounds could be regarded as an effective ingredient to bring about sleep activity associated with increased sleep time of extract (BEE) or fraction (BEE-H-11). Therefore, the water-surface activity of the extract (BEE) and fraction (BEE-H-11) of the present invention can be presumed to be due to the action of the compound in which the single compound is mixed or the compound not yet revealed.

Experimental Example  9: BEE - H2  Fraction GPCR (G- Protein Coupled receptor ) Active inhibition

Whether the BEE-H-2 fraction inhibited the activity of GPCR was determined according to the GPCRProfiler (TM) method, commissioned by Millipore Corporation (USA). As a result, there was an inhibitory effect of more than 50% on seven GPCR receptors (serotonin 1A receptor, adenosine 1 receptor, histamine 1 receptor, histamine 2 receptor, acetylcholine 2 receptor, vasopressin 1A receptor and neuropeptide Y2 receptor) ).

Experimental Example  10: Sleep mechanism of rice or rice bran extract

In order to confirm the sleeping action of the rice water extract (RWE) of Preparation Example 1 and the rice bran ethanol extract (BEE) of Preparation Example 3, 2-pyridylethyleneamine dihydrochloride, a histamine receptor agonist, Effects on the effects of

The effect of the histamine receptor antagonist on the sleep-inducing effect is inhibited by histamine receptor agonists. In this experiment, the mechanism of action of rice bran ethanol extract was demonstrated using pyrilamine maleate salt (PMS) and 2-pyridylethylamine dihydrochloride (PD) acting on histamine receptor.

(RWE) and rice bran ethanol extract (BEE) were 500 mg / kg and the antagonist PMS was 70 mg / kg, respectively, in order to investigate the effects of agonist PD on the induction effect of rice or rice bran extracts. Twenty - five minutes before oral administration of BEE and PMS, 20 mg / kg of agonist PD was administered orally before 45 minutes of hypnotic dosage (45 mg / kg), and sleep latency and sleep duration ) Were measured.

As a result of experiments, the sleeping effect of the histamine receptor antagonist PMS was completely inhibited by PD, which is an agonist, as is well known, as can be seen from FIG. 39 to FIG. Rice water extract (RWE) and rice bran ethanol extract (BEE) were also inhibited by PD, a histamine receptor agonist, like PMS.

Diazepam (DZP) was administered at a dose of 0.5 mg / kg, in order to investigate the effect of diazepam, which is one of the typical sleeping agents, on the well-known diazepam (DZP), an active agent of GABAA-benzodiazepine, BEE 500 mg / kg, antagonist PMS 70 mg / kg was given orally 45 minutes before the administration of pentobarbital (hypnotic dosage 45 mg / kg) and the histamine receptor agonist PD 20 mg / kg or GABAA - benzodiazepine antagonist Sleep latency and sleep duration were measured by intraperitoneal injection of flumazenil (FLU) 8 mg / kg 10 minutes before oral administration of DZP, BEE and PMS.

As shown in FIGS. 43 and 44, the histamine receptor agonist PD did not affect the onset time or sleeping period of diazepam (DZP), and as shown in FIGS. 38 and 39, GABAA - The benzodiazepine antagonist, flumazenil, was found to affect the sleep effect of diazepam but not the sleep effect of rice bran ethanol extract (BEE).

In the above results, rice bran ethanol extract (BEE) was found to exhibit a sleeping effect by antagonizing the histamine receptor rather than the GABAA-benzodiazepine receptor binding.

Claims (21)

A pharmaceutical composition having an antihistaminic activity characterized by containing rice, rice bran or rice husk extract as an active ingredient and acting as a histamine H1 receptor antagonist or a histamine H2 receptor antagonist and for preventing or treating sleep disorders. delete The method according to claim 1,
Wherein the prevention or treatment of the sleep disorder is for reducing the onset time, increasing the sleep duration, or increasing the sleeping surface. The method according to claim 1,
Wherein the rice, rice bran or rice hull extract is an extract of rice, rice bran or rice hull water, an organic solvent or a mixture thereof. 5. The method of claim 4,
Wherein the organic solvent is at least one solvent selected from the group consisting of lower alcohols having 1 to 6 carbon atoms, hexane, acetone, ethyl acetate, chloroform, and diethyl ether. 6. The method of claim 5,
Wherein the organic solvent is ethanol. The method according to claim 1,
Wherein the rice, rice bran or rice hull extract comprises at least one selected from the group consisting of an oil fraction of rice, rice bran or rice hull extract, a liquid fraction of rice, rice bran or rice hull extract, and a wax fraction of rice, rice bran or rice hull extract Lt; RTI ID = 0.0 &gt; antihistaminic &lt; / RTI &gt; 8. The method of claim 7,
Wherein the oil fraction of the rice bran extract is rice bran oil. The method according to claim 1,
Wherein the rice, rice bran or rice hull extract is a fraction obtained by fractionating an organic solvent extract of rice, rice bran or rice husk with a second organic solvent. The method according to claim 1,
Wherein the rice, rice bran or rice hull extract is a fraction obtained by fractionating a lower alcohol extract having 1 to 6 carbon atoms in rice bran with a second organic solvent. The method according to claim 1,
Wherein the rice, rice bran or rice hull extract is a fraction obtained by fractionating an ethanol extract of rice bran with hexane. The method according to claim 1,
Wherein the rice, rice bran or rice hull extract comprises 5 to 20 mg / g of gamma oryzanol, 0.1 to 1 μg / g of ferulic acid and 0.1 to 1 μg / g of octacosanol. A food composition for preventing or ameliorating sleep disorders through antihistamine activity, which comprises rice, rice bran or rice husk extract as an active ingredient and acts as histamine H1 receptor antagonist or histamine H2 receptor antagonist. 14. The method of claim 13,
Wherein the prevention or amelioration of the sleep disorder is for reduction of the sleeping time, increase of the sleeping time or increase of the sleeping surface. 15. The method of claim 14,
Wherein the rice, rice bran or rice hull extract is an extract of rice, rice bran or rice hull water, an organic solvent or a mixture thereof. 16. The method of claim 15,
Wherein the organic solvent is at least one solvent selected from the group consisting of lower alcohols having 1 to 6 carbon atoms, hexane, acetone, ethyl acetate, chloroform, and diethyl ether. 17. The method of claim 16,
Wherein the organic solvent is ethanol. 15. The method of claim 14,
Wherein the rice, rice bran or rice hull extract comprises at least one selected from the group consisting of an oil fraction of rice, rice bran or rice hull extract, a liquid fraction of rice, rice bran or rice hull extract, and a wax fraction of rice, rice bran or rice hull extract To &lt; / RTI &gt; 19. The method of claim 18,
Wherein the oil fraction of the rice bran extract is rice bran oil. 15. The method of claim 14,
Wherein the rice, rice bran or rice hull extract is a fraction obtained by fractionating an ethanol extract of rice, rice bran or rice husk with hexane. 15. The method of claim 14,
Wherein the rice, rice bran or rice hull extract comprises 5 to 20 mg / g of gamma oryzanol, 0.1 to 1 μg / g of ferulic acid and 0.1 to 1 μg / g of octacosanol.

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