KR20210057635A - Composition for preventing or treating tinnitus comprising extract of Castanopsis echinocarpa - Google Patents

Abstract

The present invention provides a pharmaceutical composition for preventing or treating tinnitus containing an extract or a fraction of Castanopsis echinocarpa. In addition, the present invention provides health functional food for preventing or alleviating tinnitus, containing the extract or the fraction of Castanopsis echinocarpa.

Description

Composition for preventing or treating tinnitus comprising extract of Castanopsis echinocarpa}

The present invention relates to a composition for the prevention, improvement or treatment of tinnitus comprising an extract or fraction of Castanopsis echinocarpa , which is a plant of the genus buckwheat pine.

Tinnitus is often referred to as tinnitus (perception of sound in the absence of an external source of acoustic signals). Tinnitus (耳鳴症), tinnitus, or ringing in the ears is the recognition of sounds in a person's ears without a corresponding external sound. To put it simply, sounds that do not occur outside are heard inside (Jastreboff et al., J Am Acad Audiol,. 11(3), pp162-177, 2000).

This tinnitus may appear and disappear for a while, may persist, or may be felt as a persistent sound. The pitch of the sound also varies, and usually appears in one ear, or can be heard in both ears. Statistically, about 15 to 20% of adults experience various tinnitus, of which 4% have severe tinnitus. It is also reported that 70-80% of people with hearing loss experience tinnitus (Demeester K et al., B-ENT., l7, 37-49, 2007).

Tinnitus can be divided into objective tinnitus (or objective tinnitus) that can be heard from outside and subjective tinnitus (or subjective tinnitus) that only the tinnitus patient can hear and is not heard from outside. Also, tinnitus can be classified into peripheral tinnitus and central tinnitus on the basis of differences in methods perceived by the diseased individual. Peripheral (or cochlear) tinnitus is presumed to originate from the peripheral nervous system and cochlea, and central tinnitus is presumed to originate from the auditory cortex.

Perceptual tinnitus, in which the doctor can hear the actual sound coming out of the patient's ear, can make a'click' or'tap' sound due to spasm of the muscles around the middle ear. Some people even hear the sound at the pulse. Pulsatile tinnitus may be caused by changes in blood flow or the flow of veins around the ear (Chandler JR, Laryngoscope, 93, pp892-895, 1983), but may be a subjective symptom caused by increased interest in blood flow in the ear. To date, the pathophysiological characteristics of subjective tinnitus are insufficiently understood, and the exact pathogenesis is unknown.

Tinnitus is usually a subjective phenomenon, so it is difficult to measure it with the general scientific hearing evaluation method. This was considered as a factor that did not succeed in the development of tinnitus treatment as it acted as a limiting point in animal experiments for tinnitus. In clinical practice, the evaluation of tinnitus is evaluated by scoring the subjective pain of the patient through a questionnaire. The condition is often clinically evaluated on a simple scale from'minor' to'extreme', depending on the practical difficulties such as sleep, meditation, or disruption of daily behavior. Persistent tinnitus can lead to irritability, fatigue, and sometimes clinical depression, and hallucinations may also be heard (McCombe et al., Clin Otolaryngol., 26(5), pp388-393, 2001.; Davis et al. ., Epidemiology of Tinnitus. In: Tyler R, editor. Tinnitus Handbook. San Diego: Singular Publishing Group; 2000. pp1-23).

There is no objective evaluation tool for tinnitus in clinical practice, and tinnitus is evaluated by relying on the subjective evaluation method appealed by the patient. Most of the tinnitus patients belong to subjective tinnitus, and the treatment for subjective tinnitus has been focused on rehabilitation to alleviate tinnitus. For example, a tinnitus masking method in which a sound similar to the patient's tinnitus is heard at a louder volume than the tinnitus from the outside through a hearing aid-like instrument. There are tinnitus retraining therapy, which treats tinnitus without hearing loss by listening to it. To date, no treatment for tinnitus has been approved by the US Food and Drug Administration (FDA).

Castanopsis echinocarpa (Castanopsis echinocarpa) is an evergreen broad-leaved tree belonging to the genus buckwheat. Indigenous to the tropical and subtropical regions of Korea, China, Japan, and Asia, but castanopsis ecinocarpa There is hardly any research on the material. In particular, there is a need for research and development on the material, considering that only recently research on seeds is in progress.

On the other hand, as research on the treatment or prevention of tinnitus is insufficient, the development of ingredients effective in the treatment and improvement of tinnitus needs to be preceded for related studies.

Under this background, the present inventors have tried to find an ingredient that can prevent, improve, and treat tinnitus targeting natural materials with excellent safety, and as a result, castanopsis ecinocarpa extract or fraction effectively prevents, improves, and treats tinnitus. And completed the present invention.

It is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of tinnitus comprising an extract or fraction of Castanopsis echinocarpa.

Another object of the present invention is to provide a food composition for preventing or improving tinnitus comprising an extract or fraction of castanopsis ecinocarpa.

This will be described in detail as follows. Meanwhile, each description and embodiment disclosed in the present application may be applied to each other description and embodiment. That is, all combinations of various elements disclosed in the present application belong to the scope of the present application. In addition, it cannot be considered that the scope of the present application is limited by the specific description described below.

In order to solve the above problems, one aspect of the present invention provides a pharmaceutical composition for the prevention or treatment of tinnitus comprising an extract or fraction of Castanopsis echinocarpa as an active ingredient.

In the present invention, " castanopsis echinocarpa " is an evergreen broad-leaved arboreous tree as a species belonging to the genus Castanopsis. It lives in the tropical and subtropical regions of Korea, China, Japan, and East Asia.

In the present invention, Castanopsis echinocarpa extract or fraction may be extracted or fractionated from Castanopsis echinocarpa grown in China, Vietnam or Laos.

In the present invention, "extract" refers to a resultant product such as a liquid component obtained by immersing a target substance in various solvents and then extracting it for a certain period of time at room temperature or warming state, and a solid component obtained by removing the solvent from the liquid component. . In addition, in addition to the resultant product, the extract itself, such as a dilution of the resultant, a concentrate thereof, a preparation thereof, a purified product, etc., may include all formulations of the material that can be prepared using the extract.

The extraction method for preparing the extract is not particularly limited, and may be performed according to a method commonly used in the art. As for the extraction temperature, a person skilled in the art may adopt various temperature ranges suitable for the extraction method, but is not limited thereto. In addition, the extraction time is different depending on the extraction method, and a person skilled in the art may adopt an appropriate extraction time, and may be performed once or multiple times. The extract obtained by performing the extraction with the primary extraction solvent is obtained in a liquid form from which impurities are removed by filtration according to a conventional method, or the obtained liquid extract is concentrated under reduced pressure and/or dried in a powder form according to a conventional method. You can get it. In addition, the extraction process may further include obtaining a fraction having a high content of the active ingredient, if necessary.

The Castanopsis echinocarpa extract may be prepared by extracting from Castanopsis echinocarpa according to a conventional extract preparation method. For example, castanopsis echinocarpa (Castanopsis echinocarpa) of about 1 to 20 times, preferably about 3 to 10 times the primary extraction solvent, preferably water, C ₁ to C ₄ lower It can be carried out by extraction with alcohol (eg, methanol, ethanol, propanol, isopropanol, butanol, etc.) or a mixed solvent thereof. Preferably, it was prepared by extraction with an aqueous ethanol solution. Specifically, it may be extracted with about 50 to 90% aqueous ethanol solution, more preferably about 60 to 80% ethanol aqueous solution, and even more preferably about 70% ethanol aqueous solution.

Extraction may be performed by an extraction method known in the art, for example, cold sedimentation, hot water extraction, ultrasonic extraction, reflux cooling extraction, or the like, but is not limited thereto.

The extraction temperature may be adopted by a person skilled in the art in various temperature ranges suitable for the extraction method, and may be performed at, for example, 20° C. to 100° C., but is not limited thereto. In addition, the extraction time is different depending on the extraction method, and a person skilled in the art may adopt an appropriate extraction time, but is not limited thereto, but may be performed once or multiple times in the range of about 1 hour to 10 days. The extract obtained by performing the extraction with the primary extraction solvent is obtained in a liquid form from which impurities are removed by filtration according to a conventional method, or the obtained liquid extract is concentrated under reduced pressure and/or dried in a powder form according to a conventional method. You can get it.

Before the extraction process, an enzyme treatment such as a cell wall degrading enzyme for removing the cell wall, etc., an ultrasonic treatment, a vortexing treatment, etc. may be performed, and then extraction may be performed.

In addition, the extraction process may further include obtaining a fraction having a high content of the active ingredient, if necessary. That is, after dispersing the extract obtained by extracting with the primary extraction solvent in water, extracting with an appropriate secondary extraction solvent, for example, water saturated C ₁ -C ₄ alcohol, is performed to increase the content of the active ingredient. I can.

Castanopsis echinocarpa (Castanopsis echinocarpa) of the present invention based on the total weight of the composition containing the extract as an active ingredient 0.1 to 95% by weight castanopsis echinoccarpa echinocarpa ) It is preferable to include an extract, but is not limited thereto.

In the present invention, "fraction" means a result obtained by performing fractionation in order to separate a specific component or a specific component group from a mixture containing several different constituents.

The fractionation method for preparing the fraction is not particularly limited, and may be performed according to a method commonly used in the art. For example, a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing through an ultrafiltration membrane having a certain molecular weight cut-off value, and various chromatography (separation according to size, charge, hydrophobicity or affinity) It may be a chromatographic fractionation method, and a combination thereof. In the present invention, the type of solvent used to obtain the fraction is not particularly limited, and any solvent known in the art may be used. As a non-limiting example of the fractionation solvent, water, an organic solvent, or a mixed solvent thereof may be used, and the organic solvent is an alcohol having 1 to 4 carbon atoms, a polar solvent such as ethyl acetate or acetone, hexane or dichloro. It is possible to use a non-polar solvent of methane or a mixed solvent thereof.

In the present invention, the extract or fraction may be obtained from castanopsis echinoccarpa outpost. In addition, specifically, it may be obtained from any one selected from the group consisting of leaves, stems, and combinations thereof of Castanopsis echinoccarpa, but is not limited thereto.

In the present invention, Castanopsis echinocarpa (Castanopsis echinocarpa) When the stem and leaf extracts or fractions are included together, Castanopsis echinocarpa stems and leaves are separately extracted and the combined extract or It may be a fraction, or an extract or fraction obtained by extracting the stem and leaves of Castanopsis echinocarpa together.

In the present invention, "Tinnitus" refers to the perception of sound in the absence of an external source of an acoustic signal, and may include perceptual tinnitus, subjective tinnitus, peripheral tinnitus, and central tinnitus, but is not limited thereto. Specifically, it may include subjective tinnitus caused by various causes such as noise, drugs, aging, trauma, and viruses, but is not limited thereto.

The subjective tinnitus (or objective tinnitus) refers to a tinnitus that can be heard from outside, and the subjective tinnitus (or subjective tinnitus) refers to a tinnitus that is heard only by the tinnitus patient and is not heard from outside. Also, tinnitus can be classified into peripheral tinnitus and central tinnitus on the basis of differences in methods perceived by the diseased individual. Peripheral (or cochlear) tinnitus is presumed to originate from the peripheral nervous system and cochlea, and central tinnitus is presumed to originate from the auditory cortex.

However, if it corresponds to tinnitus, it is not limited to the scope of the tinnitus of the present invention and may be included in the disease range.

The pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier, and oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories and It can be formulated in the form of a sterile injectable solution.

The pharmaceutically acceptable carriers are those commonly used in the art, such as lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like. In addition, the pharmaceutical composition of the present invention may include diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and other pharmaceutically acceptable additives, but is not limited thereto.

When the pharmaceutical composition of the present invention is formulated as an oral solid preparation, it includes tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient such as starch, calcium carbonate, It may include sucrose, lactose, gelatin, and the like, and includes, but is not limited to, lubricants such as magnesium stearate and talc.

When the pharmaceutical composition of the present invention is formulated in a liquid form for oral use, it includes suspensions, liquids, emulsions, syrups, and the like, and includes, but is not limited to, diluents such as water and liquid paraffin, wetting agents, sweetening agents, fragrances, preservatives, and the like.

When the pharmaceutical composition of the present invention is formulated for parenteral use, it includes sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories, and non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, and olives. Vegetable oils such as oils, injectable esters such as ethyloleate, and the like are included, but are not limited thereto. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used, but are not limited thereto.

Castanopsis ecinocarpa contained in the pharmaceutical composition of the present invention The dosage of the extract or fraction varies depending on the patient's condition and weight, age, degree of disease, drug type, administration route and duration, but may be appropriately selected by those skilled in the art. For example, the castanopsis echinoccarpa extract can be administered at a dose of 0.0001 to 100 mg/kg per day, preferably 0.001 to 10 mg/kg, and the administration may be administered once or several times a day. May be.

The pharmaceutical composition of the present invention can be injected into mammals such as rats, mice, livestock, humans, and fish such as zebrafish by various routes, for example, oral, intraperitoneal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injection. Can be administered by.

The pharmaceutical composition of the present invention is castanopsis ecinocarpa based on the total weight of the composition. The extract or fraction may be included in an amount of 0.01 to 95% by weight, preferably 1 to 80% by weight. In addition, castanopsis ecinocarpa contained in the pharmaceutical composition of the present invention The extract or fraction may be obtained by the same or similar method as the above-described extraction method or fractionation method, but is not limited thereto.

The pharmaceutical composition for preventing or treating tinnitus of the present invention has an excellent therapeutic effect on tinnitus by reducing abnormal behaviors caused by tinnitus.

In particular, castanopsis ecinocarpa extract or fraction has a remarkable effect on the prevention and treatment of tinnitus, and has high economical efficiency by using a material that has not been used as a medicinal or health functional food until now.

Another aspect of the present invention provides a method for preventing or treating tinnitus comprising administering the pharmaceutical composition to an individual.

The pharmaceutical composition and tinnitus are as described above.

The term "individual" of the present invention refers to all animals except humans who have or may develop tinnitus, and typically, it is beneficial by treatment with a pharmaceutical composition comprising an extract or fraction of Castanopsis ecinocarpa as an active ingredient. It may be an animal capable of exhibiting an effect, but includes without limitation any individual who has symptoms of tinnitus or is likely to have such symptoms. As described above, the tinnitus can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to an individual. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent, or may be administered in combination with an existing tinnitus therapeutic agent, and may be administered sequentially or simultaneously with the conventional therapeutic agent.

The term "administration" of the present invention means introducing a predetermined substance to a patient by an appropriate method, and the route of administration of the composition may be administered through any general route as long as it can reach the target tissue. In addition, the pharmaceutical composition of the present invention may be administered by any device capable of moving the active substance to a target tissue.

Administration of the pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent. Considering all of the above factors, it may be administered in an amount capable of obtaining the maximum effect in a minimum amount without side effects, and this can be easily determined by a person skilled in the art.

Another aspect of the present invention provides a food composition for preventing or improving tinnitus comprising an extract or fraction of castanopsis ecinocarpa as an active ingredient.

The term "improvement" of the present invention means any action in which tinnitus is improved or advantageously changed by administration of the composition of the present invention.

The castanopsis ecinocarpa, extract, fraction, and tinnitus are as described above.

The food composition of the present invention may contain conventional food additives, and the suitability as the "food additive" is, unless otherwise specified, according to the general rules and general test methods of food additives approved by the Ministry of Food and Drug Safety. It is judged according to the standards and standards for the item.

Items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigments, licorice extract, crystalline cellulose, high color pigments, and guar gum, Mixed preparations such as sodium L-glutamate preparation, an alkali additive for noodles, a preservative preparation, and a tar color preparation.

In addition, the food composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of preventing and/or improving tinnitus.

The food composition of the present invention can be used as a health functional food. The term "health functional food" refers to a food manufactured and processed using raw materials or ingredients that have functions useful for the human body according to the Health Functional Food Act, and the term "functional" refers to the structure and function of the human body. On the other hand, it means ingestion for the purpose of obtaining useful effects for health purposes such as controlling nutrients or physiological effects.

For example, the health functional food in the form of a tablet is a mixture of castanopsis ecinocarpa extract or fraction, excipients, binders, disintegrants, and other additives are granulated by a conventional method, and then a lubricant is added. Compression molding, or the mixture can be directly compression molded. In addition, the health functional food in the form of a tablet may contain a mating agent or the like, if necessary, and may be coated with a suitable coating agent if necessary.

Among the health functional foods in the form of capsules, hard capsules can be prepared by filling a conventional hard capsule with a mixture of castanopsis ecinocarpa extract or fraction, and additives such as excipients, or granular or skinned granules thereof. , Soft capsules can be prepared by filling a mixture of castanopsis ecinocarpa extracts or fractions and additives such as excipients into a capsule base such as gelatin. The soft capsule agent may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

Ring-shaped health functional foods can be prepared by molding a mixture of castanopsis ecinocarpa extract or fraction, excipient, binder, disintegrant, etc. by appropriate method, and if necessary, coating with a white sugar or other suitable coating agent, or Starch, talc, or a suitable substance can also be used to coat the ring.

The health functional food in the form of granules can be prepared in granular form by a mixture of castanopsis ecinocarpa extract or fractions, excipients, binders, and disintegrants, etc., and may contain flavoring agents, flavoring agents, etc., if necessary. have. For health functional foods in the form of granules, use No. 12 (1680 μm), No. 14 (1410 μm) and No. 45 (350 μm) sieves. It is less than 5.0% and passing through No. 45 may be less than 15.0% of the total amount.

The definitions of terms for the excipients, binders, disintegrants, lubricants, flavoring agents, flavoring agents, etc. are described in documents known in the art and include those having the same or similar functions (the Korean Pharmacopoeia commentary, Moonseongsa, Korea College of Pharmacy Council, 5th edition, p.33-48, 1989).

There is no particular limitation on the kind of the food, and examples of foods to which the castanopsis ecinocarpa extract or fraction of the present invention can be added include beverages, gums, vitamin complexes, drinks, etc., and health functions in the usual sense. Includes all food.

The food composition of the present invention may include 0.01 to 95% by weight, preferably 1 to 80% by weight, of a castanopsis ecinocarpa extract or fraction based on the total weight of the composition. In addition, castanopsis ecinocarpa contained in the food composition of the present invention The extract or fraction may be obtained by the same or similar method as the extraction method or fractionation method mentioned in the preparation of the pharmaceutical composition, but is not limited thereto.

Another aspect of the present invention provides a health functional food comprising the food composition.

The food composition and health functional food are as described above.

The present invention also provides a castanopsis ecinocarpa for use in the treatment of tinnitus. It provides a composition comprising an extract or fraction.

The present invention is also in the manufacture of a medicament for the treatment of tinnitus castanopsis echinoccarpa Provides the use of extracts or fractions.

Castanopsis echinocarpa extract or fraction according to the present invention can be effectively used in a pharmaceutical composition or health functional food for the treatment or prevention of tinnitus by inhibiting behaviors caused by tinnitus in a tinnitus-causing animal model. .

1 shows the time passed through the transparent tube in water and the number of rotations in the transparent tube when evaluating the behavior of Castanopsis echinocarpa extract in the tinnitus animal model (NOR: a control group that did not cause tinnitus, SS: tinnitus alone inducing group, CAE: castanopsis ecinocarpa extract 100mg/L treatment experimental group).
2 shows the results of the experimental group treated with the castanopsis ecinocarpa extract in the tinnitus alone inducing group and the tinnitus inducing group when evaluating the effect of the castanopsis ecinocarpa extract in a tinnitus animal model (SS: tinnitus alone inducing group, SS+CAE: Experimental group treated with castanopsis ecinocarpa extract in the tinnitus inducing group).

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited thereto.

Example 1. Castanopsis echinoccarpa in the tinnitus zebrafish model induced by salicylic acid ( Castanopsis echinocarpa ) Effect of extract

Castanopsis Echinoccarpa was purchased and used at the Overseas Biological Materials Center (https://www.ibmrc.re.kr/) operated by the Korea Research Institute of Bioscience and Biotechnology.

The castanopsis ecinocarpacastanopsis used above is of Chinese origin and is a 70% ethanol extract of castanopsis ecinocarpa extracted by 70% ethanol extraction method.

Castanopsis Echinoccarpa extract (CAE) was exposed to adult zebrafish, and the effect on tinnitus was confirmed.

Specifically, 10 adult zebrafish were put in a water tank (2L) and exposed to 1 L of 0.03% sea salt solution or 3 mM salicylic acid solution for 5 hours. After 5 hours, it was replaced with 0.03% sea salt solution or 100 mg/L CAE, and after 17 hours exposure, it was washed with 0.03% sea salt solution for 1 hour. An acrylic tube (outer diameter 25 mm, inner diameter 21 mm, total length 60 cm) was placed in a water tank (75(W) x 45(D) x 45(H) cm), and the water temperature was 28℃ by installing an underwater heater in the water tank. Kept as. Each zebrafish was passed through the acrylic tube to analyze the time and the number of rotations in the acrylic tube.

As a result, it was found that the tinnitus zebrafish exposed to 100 mg/L CAE significantly decreased the acrylic tube passage time and the number of rotations compared to the untreated control group (FIG. 1).

Example 2. Comparison of results of skinner behavioral response when evaluating the effect of castanopsis ecinocarpa extract in a tinnitus rat model induced by salicylic acid

When evaluating the skinner behavioral response in trained rats, the reaction of pressing the trigger when there is no cue tone tends to increase in animals with tinnitus. At this time, the animal misunderstands the tinnitus sound as a cue tone and shows the action of pressing the trigger, from which it can be confirmed whether or not tinnitus is caused. True positive refers to the number of times the trigger is pressed to acquire food when cue tone is provided, and false positive refers to the number of times the trigger is pressed when there is no cue tone.

Using CAE, the experimental group in which tinnitus was induced (SS) and the CAE-administered experimental group (SS+CAE) were divided into two groups, each of 5 mice, and evaluated. The SS group and CAE group used salicylic acid, which is generally used in tinnitus inducing animal models, to induce tinnitus, and 350 mg/kg was administered orally 3 hours before the test. In the SS group, only salicylic acid was administered orally, and the test was conducted in the SS+CAE group after the induction of tinnitus.The CAE study schedule was 1 day before administration (Base), and 3 days after simultaneous administration of salicylic acid (100 mg/kg). It was administered orally 1 hour and 30 minutes before.

Behavioral reactions were performed during the tinnitus induction period (1 to 3 days) and the test during the tinnitus recovery period (4 to 6 days). In the case of the SS group, salicylic acid is administered 3 hours before the test on days 1 to 3, and then water is administered 1 hour and 30 minutes before the test, and on days 4 to 6, water is administered orally only 1 hour and 30 minutes before the test. In the case of the SS+CAE group, on days 1 to 3, salicylic acid was administered 3 hours before the test, then CAE was administered 1 hour and 30 minutes before the test, and on days 4 to 6, only CAE was administered orally 1 hour and 30 minutes before the test.

As shown in FIG. 2, for the behavioral response analysis, values of a Silence activity ratio (SA ratio) and a false response rate (False positive ratio, FP ratio) were used. The SA ratio is calculated by dividing the number of reactions during the time that the cue tone was provided from the number of reactions when there is no cue tone, and the FP ratio is calculated as the ratio of the number of reactions when there is no cue tone from the total number of reactions. .

As a result, as shown in FIG. 2, it can be seen that the SS group significantly increased the SA ratio and the FP ratio compared to 0 Day, resulting in tinnitus. In the case of SS+CAE group, it was confirmed that tinnitus was suppressed by significantly decreasing the SA ratio and FP ratio compared to the SS group at 2, 3 and 4 days (*p<0.05 vs SS, **p<0.01 vs SS), the symptoms of tinnitus tended to decrease compared to the SS group, although it did not decrease significantly on the 5th day.

Example 3. Icastanopsis ecinocarpa ( Castanopsis echinocarpa ) Preparation of extract

1. Preparation of hot water extract

Castanopsis Echinoccarpacastanopsis was used as it is or after drying. 100 g of outpost was put in an extraction cloth made of non-woven fabric, and then extracted with 1,300 mL (13 times) of water in an extractor equipped with a reflux cooler at 90 to 100°C for 12 to 48 hours. After cooling this extract to room temperature (1 ~ 30 ℃), the filtrate was taken. The filtrate was combined by repeating this extraction and filtration operation twice, and the concentration of Brix (%) before concentration was measured, and then concentrated to a concentration of 15 to 25% after concentration using a vacuum rotary evaporator (Heidolph). . The concentrate was prepared with a hot water (water) extract powder using a spray dryer.

2. Preparation of ethanol extract

Castanopsis Echinoccarpacastanopsis was used as it is or after drying. 100 g of outpost was put in an extraction cloth made of a nonwoven fabric, and then extracted for 6 to 48 hours at 60 to 80°C in an extractor equipped with a reflux cooler with 1,300 mL of ethanol (13 times water). After cooling the extract to room temperature (1 ~ 30 ℃), the filtrate was taken. The extraction and filtration operations were repeated twice to combine the filtrate, measure the brix (%) concentration before concentrating, and then concentrate using a vacuum rotary evaporator (Heidolph) to achieve a brix concentration of 15 to 30%. . The concentrate was prepared in an ethanol extract powder using a spray dryer.

[Preparation Example]

1. Preparation of tablets

The extracts of Example 1 and/or Example 3 were selected according to the particle size, uniformly mixed with crystalline cellulose lactose, starch, etc., granulated together, mixed with magnesium stearate, sucrose fatty acid ester, etc., and then compressed to prepare tablets. . Constituents used in tablets and their usage are as follows.

Composition of tablets Example 1 and/or Example 3 extract 245 mg Crystalline Cellulose (KP) 225mg Lactose (KP) 12.5mg Starch (KP) 12.5mg Magnesium stearate (KP) 10mg Sucrose fatty acid ester (KP) 5mg

2. Preparation of liquid

0.15% by weight of the extract of Example 1 and/or Example 5, 10% by weight of liquid fructose, 2% by weight of honey, 2% by weight of apple concentrated fruit juice (60bx), 0.5% by weight of Guarana extract powder, 0.5% by weight of hydrated citric acid, A composition containing 0.1% by weight of sodium citrate and 0.1% by weight of taurine was prepared, and then purified water was added to prepare a liquid formulation.

Claims (8)

Castanopsis echinocarpa (Castanopsis echinocarpa) A pharmaceutical composition for the prevention or treatment of tinnitus comprising an extract or fraction.
The pharmaceutical composition of claim 1, wherein the tinnitus is any one selected from the group consisting of objective tinnitus, subjective tinnitus, peripheral tinnitus, and central tinnitus.
The method of claim 1, wherein the castanopsis ecinocarpa The extract or fraction is an extract of water, C ₁ to C ₄ of lower alcohol or a mixed solvent thereof, a pharmaceutical composition for the prevention or treatment of tinnitus.
The method of claim 3, wherein the extract or fraction is castanopsis ecinocarpa A pharmaceutical composition for the prevention or treatment of tinnitus, which is an ethanol aqueous solution extract.
Castanopsis echinocarpa (Castanopsis echinocarpa) A food composition for the prevention or improvement of tinnitus comprising an extract or fraction as an active ingredient.
The food composition of claim 5, wherein the tinnitus is any one selected from the group consisting of objective tinnitus, subjective tinnitus, peripheral tinnitus, and central tinnitus.
The method of claim 5, wherein the castanopsis ecinocarpa The extract or fraction is an extract of water, C ₁ to C ₄ of lower alcohol or a mixed solvent thereof, a food composition for preventing or improving tinnitus.
The method of claim 6, wherein the extract or fraction is castanopsis ecinocarpa A food composition for preventing or improving tinnitus, which is an aqueous ethanol extract.

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