WO2014157803A1 - Composition comprising shellfish extract as active ingredient which is for anxiety relief, improvement of convulsant, sedative action, or induction or improvement of sleep - Google Patents

Abstract

The present invention relates to a composition for a GABA_A-benzodiazepine receptor activation, which includes, as an active ingredient, an extract of at least one shellfish selected from among Mytilus edulis, Tegillarca granosa and Mercenaria stimpsoni, wherein the shellfish extract has affinity with the GABA_A-benzodiazepine receptor and is a substance acquired from a natural product, and thus does not cause side effects and secures safety; and the shellfish extract can be used, by being included as an active ingredient, in forming a pharmaceutical composition for anti-anxiety, anti-convulsant and sedation and for treating insomnia, or a food composition for anxiety relief, improvement of convulsant, sedative action and induction or improvement of sleep.

Description

Anxiety relief, spasm improvement, sedation, or sleep inducing or improving composition comprising shellfish extract as an active ingredient

The present invention relates to a composition for activating the GABA _A -benzodiazepine receptor containing shellfish extract as an active ingredient, more specifically, having an affinity for the GABA _A -benzodiazepine receptor to alleviate anxiety, improve spasm, calm It relates to a pharmaceutical composition or food composition having an action, and sleep inducing and improving effect.

Many people in modern society suffer from sleep disorders due to stress, anxiety and nervousness. According to statistics, about 15% of adults are in need of drug treatment because of insomnia caused by anxiety symptoms.There are various causes of insomnia such as stress, tension, and fear. Drugs and the like are used, but these drugs have a severe problem of side effects of forming resistance and dependence when used for a long time.

Therefore, the necessity of natural supplements as a means to replace the long-term sleepers and limiters of sleeping pills is increasing, the demand for sleep-enhancing health functional food is increasing.

In addition, GABA _A receptor is a phantomic protein that forms membrane ion channels, and is closely related to regulation of sedation, sleep, anxiety, tense muscles, convulsions, memory loss, and so on, through which GABA (gamma-aminobutyric acid) acts. Done. Many drugs, including benzodiazepines, act as anxiety agents, sedatives or sleep-inducing agents to bind to these receptors and perform their pharmacological action, and the binding of drugs or adjuvants to the benzodiazepine site enhances GABA _A receptor affinity for GABA and Increasing the intracellular influx of chlorine ions has been shown to reduce anxiety, improve spasm, sedation, and induce and improve sleep. However, they still show drug dependence and cause serious problems such as muscle relaxation and forgetfulness.

Thus, the act binds to the benzodiazepine site of the GABA _A receptor by GABA _A - benzodiazepine relax while having an affinity to the receptor using material obtained from no risk of side effects, safe natural - that sleep effects adjuvant development is urgently required It is true.

Acting on the benzodiazepine site of the GABA _A receptor to GABA _A receptor in response to GABA to activate a chloride ion channel, and chlorine is introduced into the cells of the via over-polarization of the membrane potential to slow the neuronal cellular (neuronal) activity land There have been studies on herbal and plant extracts, seaweed extracts, and the like.

Shellfish, on the other hand, is one of the most commonly used ingredients, and clam flour, along with other natural ingredients, has been used as a traditional remedy in various combinations [Biol. Rev. Camb. Philos. Soc. 85, 757-775. Benkendorff, 2010]. Shellfish powder has been reported to be used for the treatment of wound improvement, kidney disease, osteoporosis, headache, dizziness, uterine bleeding, liver disease, arthritis and skin troubles, shellfish extract has anti-cancer, antihypertensive and It is known to have an anticoagulant effect [Adv. Food Nutr. Res. 65, 153-169. De Zoysa, 2012]. For example, conopeptide isolated from Conus geographus represents a component of Ziconotide / Prialt, which has been approved by the FDA in the United States as well as Europe and is used for the treatment of chronic diseases. [Biochemistry 23, 5087-5090 . Olivera et al., 1984].

However, there have been few examples in the world of marine biomaterials, especially marine functional materials with central nervous system calming effects from shellfish.

Therefore, the first problem to be solved by the present invention is to provide a pharmaceutical composition for the treatment of anti-anxiety, anti-convulsant, sedation or insomnia containing shellfish extract.

The second problem to be solved by the present invention is to provide a food composition for anxiety relief, spasm improvement, sedation, and sleep induction and improvement comprising shellfish extract.

The present invention provides an anti-anxiety, anti-convulsant, sedation or insomnia treatment or prevention pharmaceutical composition using the shellfish extract as an active ingredient in order to achieve the first object.

According to an embodiment of the present invention, the composition may be a composition for activating GABA _A -benzodiazepine receptor.

According to one embodiment of the invention, the shellfish extract is mussel (Mytilus edulis), Cockle (Tegillarca granosa) And lilies (Mercenaria stimpsoni) Any one selected from It may be an extract of the shellfish.

According to one embodiment of the invention, the shellfish extract may be a mussel extract.

According to an embodiment of the present invention, the shellfish extract may be a shellfish extract by water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.

According to an embodiment of the present invention, the shellfish extract may be a shellfish water extract prepared by extracting for 2 to 48 hours with water of 40 ~ 100 ℃.

The shellfish extract may be a fraction obtained by re-fractionation of shellfish extract by water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof with hexane or ethyl acetate.

The present invention provides a food composition for anxiety relief, spasm improvement, sedation, and sleep induction and improvement using shellfish extract as an active ingredient to achieve the second object.

According to an embodiment of the present invention, the composition may be a composition for activating GABA _A -benzodiazepine receptor.

According to one embodiment of the invention, the shellfish extract is mussel (Mytilus edulis), Cockle (Tegillarca granosa) And lilies (Mercenaria stimpsoni) Any one selected from It may be an extract of the shellfish.

According to one embodiment of the invention, the shellfish extract may be a mussel extract.

According to an embodiment of the present invention, the shellfish extract may be a shellfish extract by water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof.

According to an embodiment of the present invention, the shellfish extract may be a shellfish water extract prepared by extracting for 2 to 48 hours with water of 40 ~ 100 ℃.

The shellfish extract may be a fraction obtained by re-fractionation of shellfish extract by water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof with hexane or ethyl acetate.

According to an embodiment of the present invention, the composition may be a health functional food formulated in any one form of tablets, powders, granules and capsules.

Shellfish extract according to the present invention has an affinity for the GABA _A -benzodiazepine receptor, the composition comprising it is effective in alleviating anxiety, improving spasm, sedation, and inducing and improving sleep, side effects because it uses a substance obtained from natural products It can ensure safety without causing.

1 is a graph showing the average price of shellfish in the last five years from 2007 to 2011. In shellfish AN, Argopecten nucleus (scallops), respectively; AP, Atrina pectinata (key shell); CS, Cyclina sinensis (permanent); ME, Mytilus edulis (mussels); MS, Mercenaria stimpsoni (lilies); MV, Mactra veneriformis ( dongle ); NC, Neptunea cumingi ( cockle ); RV, Ruditapes variegatus (crotch); SP, Saxidomus purpuratus ( patches ); TG, Tegillarca granosa .

Figure 2a is a graph showing the effect of shellfish water extract on the elevation of the mice administered the pentobarbital sleeping dose (45 mg / kg, ip), Figure 2b is a pentobarbital sleeping dose (45 mg / kg, ip ) Is a graph showing the effect of shellfish water extract on the sleep time of mice administered. Each graph shows the mean value (means ± SEM, n = 10). * Indicates a significant difference at p <0.05 compared to the control and ** at p <0.01 compared to the control (Dunnet's test). 10 mL / kg of the control group (0.5% CMC-saline), 30 mg / kg diazepam and 1,000 mg / kg of shellfish water extract were orally administered (po), and pentobarbital was administered after 45 minutes. C is a control; D is diazepam, AN, Argopecten nucleus (scallop), respectively, in shellfish; AP, Atrina pectinata (key shell); CS, Cyclina sinensis (permanent); ME, Mytilus edulis (mussels); MS, Mercenaria stimpsoni (lilies); MV, Mactra veneriformis ( dongle ); NC, Neptunea cumingi ( cockle ); RV, Ruditapes variegatus (crotch); SP, Saxidomus purpuratus ( patches ); TG, Tegillarca granosa .

Figure 3a is a graph showing the effect of mussel water extract on the elevation of the mice administered a pentobarbital (hypnotic) dose (45 mg / kg, ip), Figure 3b is a pentobarbital sleeping dose (45 mg / kg, ip) is a graph showing the effect of mussel extract on the sleep time of mice. 10 mL / kg of control (0.5% CMC-saline), 2 mg / kg of diazepam and 125, 250, 500, 1000 mg / kg of mussel water extracts were orally administered (p.o.) and pentobarbital was administered after 45 minutes. Each graph shows the mean value (means ± SEM, n = 10). * Indicates a significant difference at p <0.05 compared to the control and ** at p <0.01 compared to the control (Dunnet's test). CON is the control; DPZ is diazepam and ME is mussel extract.

Figure 4a is a graph showing the effect of mussel extract on the sleep initiation rate of mice administered pentobarbital sub-hypnotic dose (30 mg / kg, ip), Figure 4b is a pentobarbital sub -It is a graph showing the effect of mussel extract on the sleep time of the mice administered the sub-hypnotic dose (30 mg / kg, ip). 10 mL / kg of control (0.5% CMC-saline), 2 mg / kg of diazepam and 125, 250, 500, 1000 mg / kg of mussel water extracts were orally administered (p.o.) and pentobarbital was administered after 45 minutes. Each graph shows the mean value (means ± SEM, n = 10). * Indicates a significant difference at p <0.05 compared to the control group and ** at p <0.01 compared to the control group (Dunnet's test). CON is the control; DPZ is diazepam and ME is mussel extract.

Figure 5a shows the elevation time according to the administration of histamine receptor agonists (PD, 2-pyridylethylamine dihydrochloride) for the mussel water extract (ME) and the antagonist (DH, doxepin dihydrochloride) acting on the histamine receptor Figure 5b is a graph showing the sleep time according to the administration of histamine receptor agonists (PD) to the mussel water extract (ME) and antagonists (DH) acting on the histamine receptor. Controls (CON, 0.5% CMC-saline 10 mL / kg), DH (30 mg / kg), ME (1500 mg / kg) were administered orally 45 minutes prior to pentobarbital administration (hypnotic dosage 45 mg / kg). * Indicates a significant difference at p <0.05 compared to the control and ** at p <0.01 compared to the control (Dunnet's test).

Figure 6a is a graph showing the elevation time of diazepam (DZP) and mussel extract (ME) according to the administration of flumazenyl (FLU), a GABA type benzodiazepine receptor antagonist, Figure 6b is flumazenyl (FLU) It is a graph showing the sleep time of diazepam (DZP) and mussel extract (ME) according to the administration. The control group (CON, 0.5% CMC-saline 10 mL / kg), DZP (2 mg / kg), ME (1500 mg / kg) was orally administered 45 minutes prior to pentobarbital administration (hypnotic dosage 45 mg / kg), FLU (8 mg / kg) was intraperitoneally injected 10 minutes prior to oral administration of DZP, ME. * Indicates a significant difference at p <0.05 compared to the control and ** at p <0.01 compared to the control (Dunnet's test).

Figure 7 shows the elevation time effect according to the dose of the mussel water extract (ME). Elevation time when ME 500, 1000, 1500 mg / kg, DZP 6 mg / kg is administered. * Indicates a significant difference at p <0.05 compared to the control and ** at p <0.01 compared to the control (Dunnet's test).

8 shows the effect of mussel water extract (ME) on the sleep structure. Wake, nonrem sleep (NREMS) and rem sleep (REMS) times when ME 500, 1000, 1500 mg / kg and DZP 6 mg / kg are administered. * Indicates a significant difference at p <0.05 compared to the control and ** at p <0.01 compared to the control (Dunnet's test).

Figure 9a is a graph showing the change in dialysis (DPZ, 6 mg / kg) of the hourly awakening, non-remem sleep and rem sleep, Figure 9b is the hourly awakening of mussel water extract (ME, 1500 mg / kg), non-remem sleep and REM sleep It is a graph showing the change. Vehicle represents a control group.

Figure 10a is a graph showing the change in diazepam (DPZ, 6 mg / kg), average time change of each stage of non-remem sleep and rem sleep, Figure 10b is the awakening, non-rem sleep and mussel water extract (ME, 1500 mg / kg) This graph shows the average time change of each stage of REM sleep. Vehicle represents a control group, * means a significant difference at p <0.05 compared to the control (Dunnet's test).

FIG. 11A is a graph showing EEG (elctroencephalogram) power density and Delta activity at nonrem sleep of diazepam (DPZ, 6 mg / kg), and FIG. 11B is a mussel water extract (ME, 1500 mg / kg) It is a graph showing EEG (elctroencephalogram) power density and delta activity in non-rem sleep. Vehicle represents a control group, * indicates a significant difference at p <0.05 compared to the control group, and ** at p <0.01 compared to the control group (Dunnet's test).

12 is a schematic diagram illustrating a process of preparing a solvent fraction from mussel water extract.

Figure 13a is a graph showing the effect of the mussel water extract solvent fraction on the elevation time of mice administered a pentobarbital hypnotic dose (45 mg / kg, ip), Figure 13b is a pentobarbital sleeping dose (45 mg / kg, ip) is a graph showing the effect of the mussel water extract solvent fraction on the sleep time of the mice administered. 10 mL / kg of control (0.5% CMC-saline), 2 mg / kg of diazepam and water fraction (DW), hexane fraction (HX), ethyl acetate fraction (EA), butanol fraction (BT) 250 mg / Kg was orally administered (po), and 45 minutes later, pentobarbital was administered. Each graph shows the mean value (means ± SEM, n = 10). * Indicates a significant difference at p <0.05 compared to the control and ** at p <0.01 compared to the control (Dunnet's test). CON is the control; DPZ is diazepam and ME is mussel extract.

Figure 14a and 14b shows the effect of flumazenil (FLU) on the sleep induction effect of the hexane fraction of the mussel extract (ME-HX) ME-HX is 25, 50, 100, 250 mg / kg, DZP was orally administered 2 mg / kg 45 minutes prior to pentobarbital (hypnotic dosage 45 mg / kg), flumazenil (FLU) was 8 mg / kg ME-HX 250 mg / kg, DZP is a graph measured for sleep latency and sleep duration by intraperitoneal injection 10 minutes before oral administration of 2 mg / kg. * Indicates significant difference at p <0.05, ** at p <0.01 compared to the control (Dunnet's test), # at p <0.05, ## at p <0.01 This means that there is a significant difference between one group and the untreated group (Unpaired Student's t-test). NS means no significant difference.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

GABA _A receptors are associated with anxiety relief, spasms, sedation, and sleep induction and amelioration effects. Drugs that act on GABA _A receptors include agonists such as benzodiazepines, diazepam (DZP), and sol. Zolpidem and the like, and antagonists include flumazenil (FLU). As mentioned above, in the case of drugs acting on the GABA _A receptor, there is a problem that causes dependency and side effects.

In the present invention, the extract of any one or more shells selected from mussels ( Mytilus edulis ), scapula ( Tegillarca granosa) and lilies ( Mercenaria stimpsoni ) among the shell extracts act on the GABA _A -benzodiazepine receptors to alleviate anxiety, improve spasm, It was confirmed that it has a sedating effect and sleep inducing and improving effects. The sleep improvement may be an effect of reducing sleep latency, increasing sleep duration, or increasing non-REM sleep (NREMS), but the scope of the present invention is not limited thereto.

The shellfish extract is preferably a mussel ( Mytilus edulis ) extract.

The shellfish extract may be extracted using water, a lower alcohol having 1 to 4 carbon atoms or a mixture thereof as an extraction solvent. The lower alcohol may be an alcohol having 1 to 6 carbon atoms. For example, methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol or normal-butanol may be used as the lower alcohol.

For example, the preparation of the water extract may be prepared by extracting the shellfish with water of 40 ~ 100 ℃ for 2 to 48 hours.

For example, the production of ethanol extract may be prepared by extracting shellfish with 35 to 75% by weight of ethanol for 2 to 36 hours at 20 to 60 ℃, preferably 2.5 to 6 hours at 40 to 50 ℃, more Preferably it is prepared by extraction for 3 hours at 45 ℃ with 70% by weight of ethanol aqueous solution.

For example, the preparation of methanol extract can be prepared by extracting shellfish with 35 to 85% by weight of methanol for 2 to 36 hours at 20 to 60 ℃, preferably for 22 to 26 hours at 20 to 30 ℃ It can be prepared by, and more preferably, extracted with 80% by weight of methanol for 24 hours at 25 ℃.

The shellfish extract may be a fraction obtained by re-fractionation of shellfish extract by water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof into an organic solvent. The organic solvent may be one or more organic solvents selected from the group consisting of lower alcohols having 1 to 4 carbon atoms, hexane, acetone, ethyl acetate, chloroform, and diethyl ether, preferably hexane or ethyl acetate.

As used herein, the term 'extract' also includes purified products in which the extract is further concentrated, purified or separated. That is, the shellfish extract is not only obtained by using the above-described extraction solvent, but also includes a concentrate obtained by applying a purification process to it. In addition, fractions obtained by passing the extract or fraction 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), etc. The fraction obtained through the various purification methods described above is also included in the shellfish extract of the present invention.

GABA _A -benzodiazepine receptor active composition comprising the shellfish extract of the present invention as an active ingredient may be a pharmaceutical composition for the treatment of anti-anxiety, anti-convulsant, sedation and insomnia.

Pharmaceutical dosage forms of the pharmaceutical compositions for the treatment of anti-anxiety, anti-convulsant, sedation and insomnia can be used in the form of their pharmaceutically acceptable salts, and can be used alone or in combination with other pharmaceutically active compounds. Can be used in the form of a set.

In addition, the pharmaceutical composition for the treatment of anti-anxiety, anti-convulsant, sedation and insomnia according to the present invention is oral preparations such as powders, granules, capsules, capsules, suspensions, emulsions, syrups, aerosols, etc. It can be formulated and used in the form of formulations, external preparations, suppositories, and sterile injectable solutions, and can include suitable carriers, excipients or diluents commonly used in the manufacture of pharmaceutical compositions for formulation.

The carrier or excipient or diluent may be lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicide, cellulose, methyl cellulose, undetermined. And various compounds or mixtures including vaginal cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like.

When formulated, it may be prepared using conventional diluents or excipients, such as fillers, weights, binders, wetting agents, disintegrating agents, surfactants.

Solid preparations for oral administration may be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin and the like in the extract. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.

Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, preservatives, etc., in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol gelatin and the like can be used.

Preferred dosages of the pharmaceutical compositions for the treatment of anti-anxiety, anti-convulsant, sedation and insomnia according to the present invention vary depending on the condition, weight, extent of disease, drug form, route of administration and duration of the patient. Can be appropriately selected. However, for the desired effect, it may be administered at 0.0001 to 2,000 mg / kg, preferably at 0.001 to 2,000 mg / kg. Administration may be once a day or may be divided several times. However, the scope of the present invention is not limited by the above dosage.

The pharmaceutical composition for the treatment of anti-anxiety, anti-convulsant, sedation and insomnia according to the present invention can be administered to mammals such as rats, mice, livestock, humans by various routes. All modes of administration may be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

In addition, the present invention is characterized by a food composition for anxiety relief, spasm improvement, sedation, and sleep induction and improvement using shellfish extract as an active ingredient.

When the extract according to the present invention is used as an active ingredient additive of a health functional food or a general food, the extract according to the present invention may be added as it is or used with other foods or food ingredients, and may be appropriately used according to a conventional method. . The mixed amount of the active ingredient can be appropriately determined depending on the purpose of use, such as prevention, health or treatment.

In general, the extract according to the invention in the preparation of food or beverage may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less with respect to the raw material. However, in the case of long-term intake for health and hygiene or health control purposes, the amount may be below the above range, and the present invention has no problem in terms of safety in terms of using an extract from natural products. The above amount can also be used.

There is no restriction | limiting in particular in the kind of said food, The foodstuff which can add the said substance is a dairy product including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, etc. , Various soups, beverages, teas, drinks, alcoholic beverages and vitamin complexes, etc., may include all foods in a conventional sense.

The beverage food of the health functional food according to the present invention may contain various flavors or natural carbohydrates as an additional ingredient, as in general drinks. The natural carbohydrates described above may be glucose, monosaccharides such as fructose, disaccharides such as maltose, sucrose and polysaccharides such as dextrin, cyclodextrin, sugar alcohols such as xylitol, sorbitol, erythritol and the like. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL functional food according to the present invention.

In addition to the above, the food composition for anxiety relief, convulsion improvement, sedation, and sleep induction and improvement according to the present invention are various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, Protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages. In addition, the composition for improving sleep of the present invention may contain fruit flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not limited, but is generally selected in the range of 0.01 to 0.1 parts by weight relative to 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in more detail with reference to preferred examples. However, these examples are intended to illustrate the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited thereby.

Reference Example 1: GABA _A Selection of Shellfish for benzodiazepine Receptor Binding Activity

In order to develop a material exhibiting anxiety relief, convulsion improvement, sedation, and sleep induction and improving activity from shellfish, 10 kinds of shellfish that are consumed in Korea are selected and the production (unit: tons) is shown in [Table 1]. (See National Statistical Office 2012).

Table 1

division	2007	2008	2009	2010	2011	Average
ME	99,741	69,890	58,434	57,711	72,642	71,684
RV	27,459	36,302	40,393	36,248	37,929	35,666
AP	11,767	6,805	8,688	10,972	8,351	9,317
NC	-	5,592	5,799	7,242	6,913	5,109
TG	28,853	2,957	6,831	5,114	3,197	9,390
SP	3,422	2,672	1,918	1,950	2,314	2,455
CS	808	1,105	938	2,424	1,659	1,387
MV	2,604	1,657	698	1,393	1,444	1,559
MS	2,843	1,454	1,460	1,146	800	1,541
AN	564	436	398	385	454	447

Abbreviations used in Table 1 above are AN, Argopecten nucleus (scallop); AP, Atrina pectinata (key clam ); CS, Cyclina sinensis (permanent); ME, Mytilus edulis (mussels); MS, Mercenaria stimpsoni (lilies); MV, Mactra veneriformis ( dongle ); NC, Neptunea cumingi ( cockle ); RV, Ruditapes variegatus (crotch); SP, Saxidomus purpuratus ( shellfish ); TG, Tegillarca granosa .

In the last five years, mussels were the highest at 71,684 tons, followed by clam, oyster and cockle.

As a result of examining the production cost of shellfish selected for the development of calming material for marine life, the production cost of shellfish was the lowest as mussels 501 won per 1 kg, followed by Dongjuk, clam clam, and cockle [Fig. 1].

Ten types of shellfish were selected for industrial utilization in consideration of production volume and unit price. The selected shellfish are shown in [Table 2].

TABLE 2

Civilization	Scientific name	English name
Clam	Ruditapes variegatus	Manila clam
Key	Atrina pectinata	Comb pen shell
Cone	Neptunea cumingi	Gastropods
Cock	Tegillarca granosa	Cockle
Remodel	Saxidomus purpuratus	Purple washington clam
Rhythm	Cyclina sinensis	Venus clam
Dongjuk	Mactra veneriformis	Surf clam
scallop	Argopecten nucleus	Scallop
Lily	Mercenaria stimpsoni	Common orient clam
mussel	Mytilus edulis	Mussel

Reference Example 2: GABA of Oral Solution _A -Benzodiazepine Receptor Binding Activity Screening

In general, extracts contain a large amount of fat-soluble components and are often used in animal experiments by dissolving or suspending them in CMC and DMSO solutions. GABA _A -benzodiazepine receptor binding activity experiments were performed to determine whether the frequently used CMC-saline and DMSO solution itself in the preparation of extract solutions for oral administration directly affects the sedation or sleep of animals.

GABA _A -benzodiazepine receptor binding activity was measured and homogenized in 20 mL of 30 mM Tris-HCl buffer (pH 7.4, keep at 4) immediately after extraction of cerebral cortex of SD rats and sonicated for 10 seconds. After centrifugation for 15 minutes at 27,000 × g and 4 ℃ conditions, the supernatant was discarded and centrifuged again with 20 mL of buffer, this process was repeated three times. In order to remove the GABA in the brain tissue incubation in water bath at 37 ℃ for 30 minutes, centrifuged and pellets were collected and used frozen at -80 ℃. After thawing the frozen membrane, centrifuge for 10 minutes at 27,000 × g and 4 ℃, discard the supernatant, add 20 mL of 50 mM Tris-citrate buffer (binding buffer, pH 7.1, keep at 4 ℃), and centrifuge. It was. After repeating this process three times, the membrane was suspended in binding buffer at a concentration of 500 mL buffer / original tissue g and used for the evaluation of binding strength. Into a 96-well plate, add 180 µl of membrane suspension, 10 µl of each extract sample, and 10 µl of [3H] flumazenil (1 nM, final concentration) (Ro 15-1788; PerkinElmer Life and Analytical Sciences, Waltham, MA, USA). Incubation for 40 minutes in ice-bath. Thereafter, the glass fibers were collected using a filter (GF / C, Whatman). Radioactivity of the samples was measured by Tri-Carb Liquid Scintillation Analyzers (Perkin-Elmer, Shelton, CT, USA). Nonspecific binding (NSB) was determined using clonazepam (1 uM, final concentration), and the binding displacement value (%) was calculated as in Equation (1) below.

Formula (1)

Binding displacement (%) = [1-((DPM-NSB DPM) / (TB DPM-NSB DPM))] × 100

(DPM: disintegrations per minute, TB: total binding, NSB: nonspecific binding)

Three solutions (water, 0.5% CMC-saline, 10% DMSO) were orally administered (po) at a concentration of 10 mL / kg, followed by ip injection of 45 mg / kg of pentobarbital. As a result, all three solutions did not show significant difference between sleep latency and sleep duration (data not shown).

From these results, it was confirmed that any one of the three solutions did not affect the sedation or sleep-induced animal experiment. In this study, 0.5% CMC-saline was selected as the solvent used in the in vivo experiment.

Preparation Example 1 Preparation of Shellfish Water Extract

Ten scallop shellfish that are widely consumed in Korea shown in [Table 2] of Reference Example 1 (Dumpling (MV), Modified Shell (SP), Scallop (AN), Gnarled (NC), Clam (RV), Key clam (AP) ), Lily (MS), Kokmak (TG), Gamu Lac (CS), Mussel (ME) were purchased from Seoul Garak Market.

After the shells were all removed, they were washed thoroughly with water and crushed equally into 3 to 5 pieces. Distilled water 5 times (v / w) per 100 g of the raw material was added and heated at 95 ℃ for 6 hours. All extracts were filtered and concentrated under reduced pressure, lyophilized, and powdered and used in the experiment.

Experimental Example 1: GABA of Shellfish Extract _A -Benzodiazepine Receptor Binding Activity Screening

Copper porridge (MV), modified dog (SP), scallops (AN), buttocks (NC), clam (RV), key clam (AP), lily (MS), scapula (TG), lychee (CS), Mussel (ME) water extracts were used to investigate their in vivo soothing or sleep inducing effects. 10 selected candidates were orally administered (po) at a single concentration of 1,000 mg / kg, followed by intraperitoneal injection of pentobarbital (45 mg / kg, ip) to compare the sedation or sleep induction effects. [FIG. 2A] and [FIG. 2B. ].

Experimental results showed that the elevation time tended to decrease significantly ( p <0.01) in lily (MS), scapula (TG), shamrock (CS) and mussel (ME).

On the other hand, sleep time was increased to p <0.01 level in lily (MS), scapula (TG) and mussels (ME).

Mussels (ME) is a shellfish with a warm nature, it is fragile, enhances energy, and controls the intestines to enhance digestion. In addition, there is a rejuvenating effect that treats the lower back and keratin disease and drives the cold air of the stomach to enhance yang [J. Ease Asian Soc. Dietary Life 14, 187-195. Baek and Choi, 2004]. Mussels among domestic shellfish are high in production and low in price, so they are widely used as food for consumption. They are not only closely related to the national diet, but also the soothing and sleep-promoting effects found in this research project. Since it is considered to be of sufficient value, in the following experiments, mussels (ME) were selected and detailed studies were conducted.

Experimental Example 2: Sedation or Sleep Induction Effect of Mussel Water Extract in Mice Induced by Pentobarbital

1) Experimental material

Pentobarbital was purchased from Hallym Pharmaceutical Company, and diazepam (DZP), an antagonist of GABA _A -benzodiazepine receptor, was used as a positive control of sedative and sleep inducing effects.

ICR mouse (18-22 g, male) and C57BL / 6N mouse (28-30 g, male) were distributed by Coatech Co., Ltd. and were used for experiments after being adapted for one week in the breeding box for experimental animals. Animals were reared under conditions of temperature 23 ± 1 ℃, humidity 55 ± 5%, light dark cycle (light on from 09:00 to 21:00) and illuminance 3000 Lux. Feed and water were freely fed. All animals were managed according to the Guidelines for the Use of Laboratory Animals by KFRI-IACUC (Korea Food Research Institute, Institutional Animal Care and Use Committee).

2) Experiment Method

Sleep induction experiments were conducted within a period of time between 1 pm and 5 pm. Ten mice (n = 10) were used per group, and the mice were fasted for 24 hr before the experiment. All samples were prepared using 0.5% CMC-saline solution and administered orally (p.o.) 45 minutes before pentobarbital administration. The general control (control) was treated with a 0.5% CMC- saline solution at a concentration of 10 mg / kg, Diazepam (Diazepam) was used as a positive control drug to compare the sleep promoting effect of the extract as one of the typical sleeping pills. Pentobarbital is used by intraperitoneal injection (ip) at a concentration of 45 mg / kg (hypnotic dosage) after intraperitoneal injection of 30 mg / kg of sub-hypnotic dosage and 45 mg / kg of hypnotic dosage, depending on the experimental design. It became. After pentobarbital treatment, each individual was moved to an independent space to measure sleep latency and duration. Elevation time was regarded as the elapsed time from the pentobarbital to the abdominal injection to lose more than 1 min of the righting reflex, the sleep time was set to the time until the recovery again. Mice that showed no sleep behavior even after 10 minutes of pentobarbital were excluded from the experiment. The method of calculating sleep onset in the sub-hypnotic dosage of pentobarbital-treated experiment is shown in Equation (2) below.

Formula (2)

Sleep onset (%) = (No.falling asleep) / (Total No.) × 100

3) Experimental results when administering pentobarbital hypnotic dose

Mussel (ME) water extract of Preparation Example 1 125, 250, 500, oral administration (po) at a concentration of 1,000 mg / kg after pentobarbital 45 mg / kg intraperitoneal injection effect [Fig. 3a ] And [FIG. 3B].

Mussels water extract was reduced the elevation time significantly (p <0.01), 500, is increased (p <0.01) the sleep time at a concentration of 1,000 mg / kg significantly exhibited a sleep promoting effects.

4) Results of pentobarbital sub-hypnotic dose administration

The mussel (ME) water extract of Preparation Example 1 was orally administered (po) at a concentration of 125, 250, 500, 1,000 mg / kg, and pentobarbital 30 mg / kg was intraperitoneally injected to sleep onset and sleep. Sleep duration results are shown in [FIG. 4A] and [FIG. 4B], respectively.

As a result, in the pentobarbital induction experiment with sub-hypnotic concentration, the control group did not induce 100% sleep as the sleep initiation rate was 30%, while the control group treated with DZP (2 mg / kg) was 100%. All were induced to sleep.

Mussel (ME) water extracts also showed a dose-dependent increase in sleep initiation rate and sleep time, especially at concentrations of 500 and 1,000 mg / kg. Significant increases ( p <0.05) were shown, respectively, at 46.3 ± 4.9 min and 58.3 ± 2.0 min.

These results confirm that mussel (ME) water extract has a calming or sleep-inducing effect as well as diazepam.

Experimental Example 3: In Vivo Mechanism of Mussel Water Extract

1) Mechanism of action as histamine receptor antagonist

Animal experiments were performed using histamine receptor agonist, 2-pyridylethyleneamine dihydrochloride (PD). Pentobarbital (45 mg / kg) after intraperitoneal injection (ip) of 150 mg / kg PD 10 minutes prior to oral administration of 30 mg / kg of doxepin hydrochloride (DH) and 1500 mg / kg of mussel water extract (ME) kg, ip) induced sleep.

According to FIGS. 5A and 5B, the control group CON was hardly affected by PD administration, and histamine 1 receptor antagonist DH significantly sleeps due to antagonism by PD as expected. Latency was increased ( p <0.01) and sleeping time was decreased ( p <0.01). However, ME showed significant decrease in elevation time ( p <0.01) and increase in sleep time ( p <0.05) regardless of PD administration. As a result, it was confirmed that mussel extract (ME) did not act on histamine 1 receptor.

2) GABA _A -Mechanism of action as benzodiazepine activity agonist

GABA _A - the in vivo mechanism of action of benzodiazepines active efficacy of diazepam (DZP) claim (agonist) - act on benzodiazepine receptors to relax or sleep promoting mussels water extract is expected to exhibit the effect (ME) and the GABA _A Animal experiments were performed using flumazenil (FLU), a GABA _A -benzodiazepine receptor antagonist.

Flumazenyl is a water-soluble imidazobenzodiazephine, which antagonizes the competition by diazepam with GABA _A receptors in the central nervous system, and is used as an antidote when diabetic expression occurs.

Oral administration of mussel water extract (ME) at a concentration of 1,500 mg / kg (po) 10 minutes prior to intraperitoneal injection (ip) of 8 mg / kg of flumagenyl, followed by pentobarbital (45 mg / kg, ip) Induced sleep.

According to FIG. 6A and FIG. 6B, the control group CON was hardly affected by flumazenyl administration. DZP (2 mg / kg) and ME (1,500 mg / kg) showed significant decreases in elevation ( p <0.01) and increase in sleep time ( p <0.01) as expected. On the other hand DZP and ME of administration of Flu Flu horseshoe horseshoe carbonyl was due to antagonism by carbonyl significantly elevation time is increased (p <0.01) was the reduction of sleep time (p <0.01).

Taken together, these results indicate that ME contains components that act as agonists of the GABA _A receptor, such as DZP.

Experimental Example 4: sleep structure analysis of mussel water extract

1) Experiment Method

C57BL / 6N mice (28-30 g) were acclimated for 1 week and then electrode implanted surgery was performed to measure electroencephalogram (EGE) and electromyogram (EMG). Mice were anesthetized with pentobarbital (50 mg / kg, i.p.) and the head was fixed in a stereotaxic instrument. After dissection of the head subcutaneous connective tissue, Mouse EEG / EMG Headmount (Pinnacle Technology Inc, Oregon, USA) was inserted for EEG and EMG measurements. After fixing with dental dental cement and sutured. Disinfection of the surgical site and administration of antibiotics were performed for 3 days to prevent inflammation due to the surgery and a recovery period was given for 10 days. In order to adapt to the measurement environment, 0.5% CMC-saline solution used in the control group was orally administered (po) 4 days before the measurement, and then the recording device was connected to induce compliance with the experimental procedure (Fig. 3 (B)). . EEG and EMG were stabilized for 5 minutes after oral administration of samples, and then measured for 24 hours using the PAL-8200 series (Pinnacle Technology Inc, Oregon, USA). The sampling rate of EEG and EMG was set to 200 Hz (epoch time: 10 s), EEG set to 0.1-25 Hz, and EMG set the filter area of 10-100 Hz to record data. Sleep structure analysis was performed by a fast Fourier transform (FFT) algorithm, using the SleepSign program (Ver. 3.0, Kissei Comtec, Nagono, Japan). The analysis results were divided into wake, REM sleep (rapid eye movement, theta band: 6-10 Hz), and NREM sleep (non-rapid eye movement, delta band: 0.65-4 Hz). Sleep latency was set as the time taken for NREM sleep in 10 s epoch units to continuously appear more than 12 times.

2) Experiment result

The sleep-promoting effect of mussel water extract (ME) was compared with the well-known sleeping agent diazepam (DZP). DZP is a benzodiazepine (BZD) that acts on the GABA _A -BZD receptor [PNAS. 98, 6464-6469. Tobler et al, 2001.

As shown in FIG. 7, the mussel water extract (ME) did not reduce the elevation time at the 500 mg / kg concentration, but the elevation time was significantly (p <0.05) at the 1000 mg / kg and 1500 mg / kg concentrations. There was also a significant decrease (p <0.01) of elevation time in the DZP 6 mg / kg concentration.

In addition, the non-lamb sleep time of the mussel extract (ME) did not increase as much as 6 mg / kg of DZP, but increased concentration-dependently, and showed no significance at 500 mg / kg concentration and 34.1% at 1000 mg / kg and 1500 mg / kg concentration, respectively. And a significant (p <0.05) increase of 32.9% (see FIG. 8).

Changes in nonremnant sleep time per hour during the measurement of EEG in C57BL / 6N mice after oral administration of mussel hydrothermal extract (ME) 1,500 mg / kg and DZP 6 mg / kg, respectively, are shown in FIGS. 9A and 9B. Indicated. ME showed a significant (p <0.05) increase in nonrem sleep for the first hour after oral administration, and DZP showed a significant increase in nonrem sleep for 6 hours than the control group. These results showed that ME induces natural sleep after sleep-promoting effect in the early stage of sleep, while DZP showed sleep-promoting effect in early stage of sleep, but the effects persisted to confirm typical side effects of sleeping pills.

To better understand the sleep-promoting effects of mussel hydrothermal extracts, the mean duration and EEG power density of each sleep stage were determined.

According to FIGS. 10A and 10B, the ME and DZP showed 47.6% and 69.2% decreases in mean duration of wake, respectively, but there was no change in non-REM and REM sleep. Reducing the mean time of awakening without changing ME and non-REM sleep suggests that ME has reduced the duration of awakening [Mol. Nutr. Food Res. 56, 304-308. Masaki et al., 2012].

Changes in delta activity during nonrem sleep are a physiological indicator of sleep quality and depth [Behav. Brain Res. 210, 5456. Huang et al., 2010]. DZP has also been reported to increase nonremem sleep and decrease delta power during nonremem sleep [PNAS. 101, 3674-3679. Kopp et al., 2004]. These typical DZP properties are also evident in this study. As expected, DZP significantly reduced delta activity during non-remem sleep (p <0.01) (see FIG. 11A), and no significant change in ME's delta activity (see FIG. 11B). These results indicate that ME increases NREM sleep without affecting physiological sleep. The ideal sleeping agent is a drug that has fast sleep time, maintains sleep, and does not change physiological sleep [Sleep. 31, 259-270. Alexandre et al., 2008]. Therefore, mussel water extract (ME) of the present invention is believed to be able to promote safer and more natural sleep than BDZ and non-BDZ drugs with side effects.

Experimental Example 5: Soothing or Sleep Inducing Effect of Mussel Extract Fraction

1) Preparation of Mussel Extract Fraction

Mussels were extracted in water at 95 ° C. for 6 hours. The extract was filtered and the remaining one was extracted three more times in the same way. The obtained filtrates were combined and concentrated under reduced pressure to obtain a water extract. 7 L of H ₂ O was added to the obtained water extract, and the mixture was extracted with n- hexane (27 L × 2) / H ₂ O (27 L), and H ₂ was extracted. O was again extracted with EtOAc (27 L × 2) and H ₂ O was extracted with n- BuOH (25 L × 2). Each layer was concentrated under reduced pressure to obtain a hexane fraction (ME-HX), an ethyl acetate fraction (ME-EA), a butanol fraction (ME-BT) and a water fraction (ME-DW) (see FIG. 12).

2) Results of sedation or sleep induction effect of mussel extract fraction

In order to confirm the sedation or sleep induction enhancement effect of the mussel extract fraction orally administered (po) at a concentration of 250 mg / kg (45) each pentobarbital 45 mg / kg intraperitoneally injected to change the change in sleep time and sleep time Confirmed.

13A and 13B, the ME-DW fractions showed no sedative or sleep enhancing effect, and significant elevations in fractions ME-HX, ME-EA and ME-BT except ME-DW fractions. A decrease in time ( p <0.01) was shown. In sleep time, the sleep-promoting effect of ME-HX and ME-EA fractions was significantly increased ( p <0.01), and the sedation and sleep-promoting effects of ME-HX fractions were higher than those of ME-EA fractions. Confirmed.

Experimental Example 6: Mussel Extract Hexane Fraction (ME-HX) in vivo Mechanism of Action

Sleep dose of mussel hexane fraction (ME-HX) to investigate pentobarbital induced sedation and sleep promotion effects, oral administration of ME-HX at concentrations of 25, 50, 100 and 250 mg / kg Sleep induction effect by the slope (45 mg / kg, ip) is shown in Figure 14a and 14b.

According to FIGS. 14A and 14B, significant elevation decreases ( p <0.01) and increased sleep times ( p <0.01) were observed in 250 mg / kg of ME-HX fractions, 25, 50 and 100 At concentrations of mg / kg no sedative or sleep-promoting effects were seen.

Therefore, the ME-HX fraction could be considered to have a sedative and sleep-promoting effect at the concentration of 250 mg / kg, and thus the ME-HX fraction, which is expected to have a sleep-promoting effect by acting on the GABA _A -benzodiazepine receptor and GABA _a - benzodiazepine GABA _a receptor agonist to determine the in vivo mechanism of action of diazepam (DZP) - using the benzodiazepine antagonist chain flu horseshoe carbonyl (fLU) were conducted animal experiments.

Oral administration of the ME-HX fraction at a concentration of 200 mg / kg (ip) 10 minutes prior to intraperitoneal injection (ip) of 8 mg / kg of flumagenyl followed by sleep with pentobarbital (45 mg / kg, ip) Induced. Experimental results showed that the control group (CON) was hardly affected by flumazenyl administration. The DZP (2 mg / kg) and ME-HX fractions showed significant decreases in elevation ( p <0.01) and sleep times ( p <0.01) as expected. On the other hand flu horseshoe in the DZP (2 mg / kg), and ME-HX fraction treated with carbonyl flu horseshoe due to antagonism by carbonyl significantly the elevation time increase (p <0.01) was the sleep time reduction (p < 0.01) (see FIG. 14A and FIG. 14B).

Taken together, these results indicate that the ME-HX fraction contains components that act as agonists of the GABA _A receptor, such as DZP. As a result, the active ingredients of the n- hexane fraction of mussels are derived from the GABA _A receptor. It was confirmed that it acts as an agonist.

Experimental Example 7: Statistical Analysis

The results of all experiments were expressed as mean and standard error (means ± SEM), and statistical comparison of each group and control was evaluated by One-way-analysis of variance (ANOVA) by Dunnet's test. The significance level was marked with an asterisk indicating the results showing significance at p <0.05 (*), P <0.01 (**), and p <0.001 (***) levels. Data comparison between the two groups was analyzed by Unpaired Student's t-test, and statistically significant differences between the two groups at the levels of p <0.05 (#), p <0.01 (##), and p <0.001 (###) Indicated that there is. Statistical analysis program was performed using Prism 5.0 software.

Hereinafter, a preparation example of a composition containing an extract of the present invention will be described, but the present invention is not intended to be limited thereto but only to be described in detail.

Formulation Example 1 Preparation of Powder

20 mg of mussel water extract (ME) powder of Preparation Example 1

Lactose 100 mg

Talc 10 mg

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

Formulation Example 2 Preparation of Tablet

10 mg of mussel water extract (ME) powder of Preparation Example 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

Formulation Example 3 Preparation of Capsule

10 mg of mussel water extract (ME) powder of Preparation Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

10 mg of mussel water extract (ME) powder of Preparation Example 1

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO _4, 12H ₂ O 26 mg

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

Formulation Example 5 Preparation of Liquid

20 mg of mussel water extract (ME) powder of Preparation Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve, the lemon flavor is appropriately added, the above components are mixed, the purified water is added, the whole is adjusted to 100 by adding purified water, and then filled into a brown bottle and sterilized. To prepare a liquid solution.

Formulation Example 6 Preparation of Health Functional Food

1,000 mg mussel water extract (ME) powder of Preparation Example 1

Vitamin Mixture

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinamide 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5 mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

15 mg potassium monophosphate

Dicalcium Phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is a composition that is relatively suitable for the health functional food, the composition is mixed in a preferred embodiment, but the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health functional food manufacturing method. Then, the granules may be prepared and used for preparing the nutraceutical composition according to a conventional method.

Formulation Example 7 Preparation of Functional Beverage

1,000 mg mussel water extract (ME) powder of Preparation Example 1

Citric Acid 1,000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 mL of purified water

After mixing the above components according to the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated Used to prepare functional beverage compositions of the invention.

Although the composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

Shellfish extract according to the present invention has an affinity for the GABA _A -benzodiazepine receptor, the composition comprising it is effective in alleviating anxiety, improving spasm, sedation, and inducing and improving sleep, side effects because it uses a substance obtained from natural products It is possible to secure safety without causing, and is very useful as a food composition such as pharmaceutical composition or health functional food.

Claims (15)

1. mussel(Mytilus edulis), Cockle (Tegillarca granosa) And lilies (Mercenaria stimpsoni) Any one selected from Anti-anxiety, anti-convulsant, sedation or insomnia treatment or prevention pharmaceutical composition comprising the extract of the shellfish as an active ingredient.
2. The method of claim 1, wherein the shell extract is anti-anxiety, anti-convulsant, sedation or insomnia treatment or prevention pharmaceutical, characterized in that to activate the GABA (gamma-aminobutyric acid) type A-benzodiazepine receptor Composition.
3. The pharmaceutical composition for treating or preventing insecure, anti-convulsant, sedation or insomnia according to claim 1, wherein the shellfish is mussel ( Mytilus edulis ).
4. [Claim 2] The anti-anxiety, anti-convulsant, sedative or insomnia treatment or prevention according to claim 1, wherein the shell extract is a shell extract by water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. Pharmaceutical composition for.
5. The anti-anxiety, anti-convulsant, sedation or insomnia treatment or prevention according to claim 4, wherein the shellfish extract is a shellfish water extract prepared by extracting for 2 to 48 hours with water at 40 to 100 ° C. Pharmaceutical composition for.
6. [5] The anti-anxiety and anticonvulsant according to claim 4, wherein the shellfish extract is a fraction obtained by re-fractionation of the shellfish extract by water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof into an organic solvent. ), A pharmaceutical composition for treating or preventing sedation or insomnia.
7. The method of claim 6, wherein the organic solvent is at least one solvent selected from the group consisting of lower alcohols having 1 to 4 carbon atoms, hexane, acetone, ethyl acetate, chloroform, and diethyl ether. anti-convulsant), pharmaceutical composition for the treatment or prevention of sedation or insomnia.
8. 8. The pharmaceutical composition for treating or preventing insecure, anti-convulsant, sedation or insomnia according to claim 7, wherein the organic solvent is hexane or ethyl acetate.
9. mussel(Mytilus edulis), Cockle (Tegillarca granosa) And lilies (Mercenaria stimpsoni) Any one selected from Anxiety relief, convulsion improvement, sedation, or sleep inducing or improving food composition comprising the extract of the shellfish as an active ingredient.
10. 10. The food composition of claim 9, wherein the shellfish extract activates GABA (gamma-aminobutyric acid) type A-benzodiazepine receptor.
11. The food composition of claim 9, wherein the shellfish is a mussel ( Mytilus edulis ).
12. The method of claim 9, wherein the shell extract is an anxiety relief, spasm improvement, sedation, or sleep inducing or improving food composition, characterized in that the shell extract by water, lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof. .
13. The method of claim 12, wherein the shell extract is an anxiety relief, spasm improvement, sedation, or sleep inducing or improving food composition, characterized in that the shell water extract prepared by extracting for 2 to 48 hours with water of 40 ~ 100 ℃ .
14. The method of claim 12, wherein the shell extract is an anxiety relief, spasm improvement, calming, characterized in that the fraction re-fractionated with water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof shellfish extract with hexane or ethyl acetate Food composition for action, or sleep induction or improvement.
15. 10. The method of claim 9, wherein the composition is a dietary supplement formulated in any one form of tablets, powders, granules and capsules anxiety relief, spasm improvement, sedation, or food composition for inducing or improving sleep.

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