KR101906720B1 - Composition comprising the combination extract of Lycium chinensis, Glycyrrhiza urlaensis FISCH, Foeniculum vulagare MILL, Glycine max MERR, and Pueraria thunbergiana BENTH for preventing and treating menopause-related disease and depression - Google Patents

Abstract

The present invention relates to a composition for the prophylaxis and treatment of female menopausal diseases and depression comprising, as an active ingredient, a combination herbal extract of Gugija, licorice, fennel, soybean and grenache. In detail, the combination extract of Gujia, Licorice, Fennel, Soybean, and Pueraria of the present invention strongly suppresses depression as compared with a conventional antidepressant, based on results of antidepressant effects of female menopause in a forced swimming test and a tail suspension test of mice. Therefore, the composition can be used in pharmaceutical compositions and health functional foods useful for prevention and treatment of the female menopausal diseases including depression.

Description

The present invention relates to a composition for prevention and treatment of female menopausal diseases and depression, which comprises a combination of a herbal extract, a licorice, a fennel, a fennel, a soybean, MERR, and Pueraria thunbergiana BENTH for < RTI ID = 0.0 > and &

The present invention relates to a composition for the prophylaxis and treatment of female menopausal diseases and depression comprising, as an active ingredient, a herbal medicine extract of a combination of Gugija, licorice, fennel, soybean and grenache.

[Patent Document 1] Cho YJ, et al., Journal of Korean medical science ., 26 (2), pp279-83, 2011

[Literature 2] Arborelius L, et al., The Journal of endocrinology. , 160 (1), pp 1-12, 1999.

[Document 3] Brady LS. et al., T he Journal of clinical investigation. 87 (3), pp 831-7, 1991

[Literature 4] Butterweck V. et al., Molecular psychiatry ., 6 (5), pp547-64, 2001

[Literature 5] Graeff FG. et al., Pharmacology, biochemistry, and behavior ., 54 (1), 129-41, 1996

[Literature 6] Garakani A. et al., Journal of affective disorders . 2012.

[Literature 7] Rovin ML et al., Neuropeptides . 46 (2), pp81-91, 2012

[Literature 8] Falowski SM. et al., Neurosurgery . 69 (6), pp1281-90, 2011

[Literature 9] Paxinos G. et al., Journal of neuroscience methods . 13 (2), pp139-43, 1985

The term "climacteric" refers to a type of endocrine syndrome that is characterized by a general and gradual decrease in ovarian function, resulting in a decrease in physiological function and sexual function. The term "climacteric" refers to a period of menopause which is a permanent stoppage of menstruation (menopause) will come. Postmenopausal women may have acute chronic symptoms due to changes in hormones, such as decreased estrogen production, follicular stimulating hormone (FSH), and elevated luteininzing hormone (LH). For example, symptoms due to vascular changes such as facial flushing, tachycardia, sweating, and headache; Symptoms due to musculoskeletal, joint pain, back pain, and other musculoskeletal changes: Symptoms due to urinary genital changes such as urinary frequency and urinary incontinence: Symptoms of changes in the brain nervous system such as memory loss, depression, And the like.

Postmenopausal estrogen production sites are the ovarian matrix, adrenal gland, subcutaneous fat, and are converted to estrone by the orientation of androstendione. In menopause, a decrease in estrogen secretion below physiologically necessary level causes various symptoms such as facial flushing, genital atrophy, dysuria, osteoporosis, lipid metabolism disorder, depression, insomnia, skin aging, (Han et al., Reproductive Sci, 2016, 23, 670). In particular, hot flushes can cause sweating, flushing, palpitation, anxiety, irritability and cold sweating (Vered et al., The Lancet, 2002, 360; 1851). Menopausal syndrome is not only a decline in physical function but also a psychological and social contraction.

Hormone replacement therapy and nonsteroidal remedies have been developed as therapeutic therapies for improving the symptoms of women's menopausal symptoms. Among them, estrogen replacement therapy is known to be the most effective method. However, the use of drugs for a long period of time according to the therapeutic regimen may cause endometrial cancer, breast cancer, hypertension, thrombosis, biliary stones, and also may cause a mild bleeding, and owing to side effects such as ovarian hyperstimulation, There is a problem that it can not be taken without. (Korean Patent No. 10-0419121)

Currently, many drug therapies have been developed for the improvement and treatment of these menopausal disorders, and hormone replacement therapy has been widely used. These hormone replacement therapies include combined / sequential estrogen and progestin therapy, continuous / combined estrogen and progestin therapy, and estrogen alone.

However, these alternative therapies are known to increase the incidence of breast cancer, including endometrial cancer and heart disease, due to excessive estrogen. Recently, there has been a growing interest in safer vegetable estrogen.

Depression is a type of psychiatric disorder that is lost in despair and loses motivation and becomes obsessed with incompetence, isolation, impatience, guilt, and suicidal impulse. It is accompanied by somatic symptoms such as dyspepsia, headache, insomnia, constipation, diarrhea and loss of libido.

Depression is highly prevalent worldwide, is accompanied by a variety of dysfunctions, and because sociocultural factors have a significant impact on the manifestation of depression and the health-seeking pattern, it is necessary to establish measures, including optimal mental health policies, For this, precise and reliable epidemiological data is essential rather than anything.

In addition, Korea is experiencing many changes such as nuclear family, personalization, rapid economic growth, rapid aging, and suicide rate is already the number one place in OEDC countries in 2010, and the importance of depression is emphasized more than ever.

Until now, the mechanism of depression has not been clearly elucidated, and the lack of serotonin, norepinephrine, and dopamine, which are monoamine neurotransmitters in central nervous system synapses, is considered as the most likely hypothesis. Thus, most antidepressants have a pharmacological action that increases the concentration of the monoamine ganglioside substance in the central nervous system synapse.

Representative depressive therapies currently in use include benxodiazepine, diazepam, oxazepam, lorazepam, alprazolam, helazepam, clonazepam, and the like. These drugs are also used as sedatives and sleep inducers.

Depression has been reported to cause depression in addition to serotonin due to the loss of neurotransmitters of dopamine and epinephrine. Serotonin regulates the excess secretion of aggressive norepinephrine, addictive endorphins and dopamine (7: Rovin ML. et al., Neuropeptides . 46 (2), pp81-91, 2012). Those who lack serotonin for a long time seem to be easily aggressive or prone to emotional excitement. Tyrosine hydroxylase (TH) is used to indirectly detect the expression of dopamine and norepinephrine as a rate-limiting enzyme of dopamine and norepinephrine (Falowski SM et al., Neurosurgery 69 (6), pp1281-90, 2011).

Gugija refers to the mature fruit of Lycium chinensis, a vine shrub belonging to Solanaceae. It contains substances such as carotene, nicotinic acid and the like, and contains glucose and cholesterol lowering effects, Blood pressure lowering action, etc. (Information Sourcing, Illustrated Dictionary of Korean Medicine, Yeongrim, pp826-828, 1998).

Licorice refers to the roots and rootstocks of the perennial herb licorice (Glycyrrhiza urlaensis FISCH) belonging to the leguminosae and contains substances such as gycyrrhizin, liquiritigenin, liquiritin and the like It is known to have a detoxifying action, a deep-sea gut action, a gastric secretion-inhibiting action, and a liver protective action (Information Sourcing, Dietary Encouragement, Yonglim, pp684-687, 1998).

Fennel refers to the seed of the perennial herb Fenniculum vulgare MILL belonging to the Umbelliferae and contains an essential oil such as anethole, fenchone, linene and the like, , And antimicrobial activity. (Information, Outline, and Illustrated Dictionary, Yeonglim, pp424-425, 1998).

Soybean refers to black seeds of Glycine max MERR, an annual herb belonging to Leguminosae, which contains substances such as protein, fat, carbohydrate, caotene and nicotinic acid, It is known to have a function (Information Extension, Illustrated Dictionary of Contemplation, Younglim Corp, pp682-683, 1998).

Pueraria thunbergiana BENTH is a perennial plant belonging to the genus Leguminosae, which is called Puerarin, puerarin xylose, daidzein, sitosterol sitosterol), and it has been known to have a blood flow increasing action, a perspiration action, and a fever-releasing action (Information Sourcing, Diagrams of Dietary Encouragement, Young Lim, pp. 704-706, 1998).

However, none of the above references discloses or discloses the efficacy of the combination extract of Ganoderma lucidum, licorice, fennel, soybean, and grenache as an antidepressant.

Accordingly, the present inventors have searched natural products for the development of a new female menopausal disease treatment and antidepressant drugs from natural products, and found that a combination extract of Gugija, licorice, fennel, soybean, It was confirmed that the immobility duration was significantly reduced in the concentration-dependent manner in the Fored swimming test and the tail suspension test, thereby completing the present invention.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of female menopausal diseases, which comprises an extract of a combination of Gugija, licorice, fennel, soybean, and bulgaria as an active ingredient.

The present invention also provides a health functional food for preventing and ameliorating female menopausal diseases, which comprises an extract of a combination of Gugija, licorice, fennel, soybean, and bulgaria as an active ingredient.

Combination herbal extracts as defined herein include water containing purified water, C1 to C4 lower alcohols or mixtures thereof, preferably water-soluble extracts.

The preferred combination weight of the combined herbal extracts as defined herein is the weight parts (w / w) of 1 to 10 grams of licorice, 1 to 10 licorice, 1 to 10 of fennel, 1 to 10 of soybean and 1 to 20 of propolis, (W / w), more preferably 1 to 5, licorice 1 to 4, fennel 1 to 10, licorice 1 to 5, fennel 1 to 5, soybean 1 to 5 and pomegranate 1 to 10 To 2 parts by weight of soybean flour, 1 to 5 parts by weight of soybean flour, and parts by weight (w / w) of flour of 1 to 10 parts by weight.

Specifically, the female menopausal disease as defined herein includes libido, hypogonadism, hypogonadism, sexual dyslipidemia, sexual maturity and the like, thereby causing nervousness, emotional instability, depression, dizziness, hot flashes, sweating, A reduction in work capacity, a decrease in physical strength, a decrease in athletic ability, a decrease in body size, a skin aging, and an increase in visceral fat, more specifically, nervousness, emotional disturbance, or depression.

Hereinafter, the present invention will be described in detail.

Combination herbal extracts of the present invention can be obtained as follows.

The combined herbal medicine extract of the present invention is obtained by washing the herbal medicine ingredients gugija, licorice, fennel, soybean, and gangrene each with water and shredding after shrimp treatment to about 1 to 20 times, preferably about 2 to 10 times Preferably at a temperature of from 0 to 120 ° C, preferably from 20 to 100 ° C, for a period of from about 1 hour to 1 day, preferably from 2 hours to 1 hour, After 8 hours of extraction, the extract is filtered after cooling, and the filtrate is lyophilized to obtain the individual extracts, which are then subjected to extraction by hot water extraction, cold extraction, reflux cooling extraction, ultrasonic extraction or supercritical extraction, And the individual extracts are mixed at a predetermined mixing ratio to obtain the combined herbal medicine extract of the present invention.

The present invention provides a pharmaceutical composition and a health functional food for the prevention and treatment of female menopausal disease, which comprises the combination extract obtained by the above-mentioned production method as an active ingredient.

Forced swimming test and Tail suspension test, which are in vivo search methods of the antidepressant drug efficacy of the combination extract of the present invention, showed that the extract was immobilized in a concentration- (Immobility duration), which is useful for the prevention and treatment of female menopausal disease.

The pharmaceutical composition containing the extract of the present invention contains 0.1 to 50% by weight of the extract, based on the total weight of the composition.

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

Since the extract of the present invention has little toxicity and side effects, it can be safely used for long-term use for preventive purposes.

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

The composition of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions, Examples of carriers, excipients and diluents that can be included in the composition containing the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions, and syrups. Various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. may be included in addition to water and liquid paraffin which are simple diluents used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.5 g / kg to 5 g / kg per day, preferably 1 g / kg to 3 g / kg per day. The administration may be carried out once a day or divided into several doses. Accordingly, the dosage is not limited in any way to the scope of the present invention.

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

Also, the present invention provides a health functional food or a health supplement food for preventing and improving female menopausal disease, which comprises an extract of Ganoderma lucidum, Licorice, Fennel, Soybean and Pigment Extract as an active ingredient.

Also, the present invention provides a health functional food or a health supplement food for preventing and improving female menopausal disease, which comprises an extract of Ganoderma lucidum, Licorice, Fennel, Soybean and Pigment Extract as an active ingredient.

Accordingly, the present invention also provides a food and food additive for preventing and improving female menopausal disease, which contains an extract of Ganoderma lucidum, licorice, fennel, soybean, and bulgaria as an active ingredient.

The composition containing the extract of the present invention can be used variously for medicines, foods and beverages for prevention and improvement of depression. Examples of the foods to which the extract of the present invention can be added include various foods, beverages, gums, tea, vitamin complexes, health supplements and the like, and they can be used as powders, granules, tablets, capsules or beverages have.

The extract of the present invention can be added to food or beverage for the purpose of preventing and improving depression. At this time, the amount of the extract in the food or beverage is generally from 1 to 5% by weight of the total food weight of the health food composition of the present invention, and the health beverage composition is preferably 0.02 to 10 g based on 100 ml, Can be added at a ratio of 0.3 to 1 g.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient in ordinary beverages, in addition to containing the above extract as an essential ingredient in the indicated ratio. Examples of the above-mentioned natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose such as maltose and sucrose and polysaccharides such as dextrin and sludge dextrin Conventional sugars and sugar alcohols such as xylitol, sorbitol, and erythritol. (Taurine, etc.), stevia extract (e.g., glyburide), and synthetic flavorings (saccharin, aspartame, etc.) are advantageously used as flavorings other than those described above . The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 mL of the composition of the present invention.

In addition to the above-mentioned composition, the composition of the present invention can be used as a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and intermediates (cheese, chocolate etc.), pectic acid and its salts, Salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the compositions of the present invention may contain flesh for the production of natural fruit juices and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

The combination extract of the present invention showed antidepressant effects by the forced swimming test and the tail suspension test of the mice, and as a result, the depression was suppressed more strongly than the existing antidepressant drugs, And can be used in pharmaceutical compositions and health functional foods useful for the prevention and treatment of female menopausal diseases such as non-harmful depression.

Figure 1 shows the effect of a drug on a forced swimming test;
Figure 2 shows the effect of the drug on the tail suspension test;
Figure 3 shows the effect of the drug on the level of oxytocin
Figure 4 shows the effect of the drug on the level of corticosterone;
Figure 5 shows the effect of the drug on estrogen levels.

Hereinafter, the present invention will be described in detail by reference examples, examples and experimental examples.

However, the following Reference Examples, Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Reference Examples, Examples and Experimental Examples.

Example  1. Preparation of drug combination extract

1-1. Manufacture of dried Gugija

Prepare Gugija (Domestic product, Daehan Pharmaceutical) which passed the quality inspection and prepare 50kg of the prepared gugije precisely weighed, put it in the extraction tank (Daesung Machinery), put the constant of 10 times volume of Gugija sample into the extraction tank, For 4 hours, and then filtered and concentrated (soft X-ray) until the concentration in the concentrator (Daesung Machinery) was 60 Brix or more. The dried gugija extract obtained by putting the soft-drink extract in a vacuum dryer (Daesung Machine) at a temperature of 60 ° C and a degree of vacuum of -60 mmHg for 9 hours under reduced pressure was pulverized with a grinder (Daesung Machinery, DS-PM-100) (Yield: 24.7%).

1-2. Manufacture of dried licorice extract

Prepare a licorice (Uzbekistan, Daehan Pharmaceutical) that has passed its own quality inspection and weigh exactly 50kg of prepared gugija, put it in an extraction tank (Daesung Machinery), put a constant of 10 times volume of gugija sample in the extraction tank, C for 4 hours, and then filtered and concentrated (soft X-ray) until the concentration in the concentrator (Daesung Machinery) was 60 Brix or more. The dried licorice extracts obtained by putting the soft-drink extracts in a vacuum drier (Daesung Machine) at a temperature of 60 ° C and a vacuum degree of -60 mmHg for 9 hours were pulverized with a pulverizer (Daesung Machinery, DS-PM-100) ≪ / RTI > (Yield: 12.9%).

1-3. Manufacture of dried fennel

(50kg) of the prepared fennel is weighed accurately, placed in an extraction tank (Daesung Machinery), and a constant 10 times volume of the sample volume of the gugija is put into the extraction tank, For 4 hours, and then filtered and concentrated (soft X-ray) until the concentration in the concentrator (Daesung Machinery) was 60 Brix or more. The dried fennel extract obtained by putting the soft-drink extract in a vacuum dryer (Daesung Machine) at a temperature of 60 ° C and a degree of vacuum of -60 mmHg for 9 hours under reduced pressure was pulverized with a pulverizer (Daesung Machinery, DS-PM-100) (Yield: 8.2%).

1-4. Manufacture of soybean dry extract

Prepare the soybeans (domestic and Daehan Pharmaceuticals) that have passed the quality inspection and weigh 50kg of the prepared soybeans exactly after weighing, put the constant volume of 10 times volume of the soybean sample into the extraction tank (Daesung Machinery) For 4 hours, and then filtered and concentrated (soft X-ray) until the concentration in the concentrator (Daesung Machinery) was 60 Brix or more. The dried soybean extract obtained by putting the soft-drink extract in a vacuum dryer (Daesung Machine) at a temperature of 60 ° C and a degree of vacuum of -60 mmHg for 9 hours under reduced pressure was pulverized with a pulverizer (Daesung Machinery, DS-PM-100) (Yield: 7.5%).

1-5. Manufacture of Drying Drying Extract

Prepare pupae (domestic, Daehan Pharmaceutical) that passed the quality inspection and prepare 50kg of the prepared pupa weighed exactly after weighing and put the constant volume of 10 times volume of the pupa into the extraction tank (Daesung Machinery) and put it in the extraction tank at 80-100 ℃ For 4 hours, and then filtered and concentrated (soft X-ray) until the concentration in the concentrator (Daesung Machinery) was 60 Brix or more. The dried soft mushroom extract obtained by putting the soft mushroom extract in a vacuum dryer (Daesung Machine) at a temperature of 60 ° C and a vacuum degree of -60 mmHg for 9 hours was pulverized with a pulverizer (Daesung Machinery, DS-PM-100) (13.1%) < RTI ID = 0.0 >

1-6. Drug combination manufacturing

(3) Fennel Drying Extract 5.0%; (4) Soybean Dry Extract 23.75%; (3) Fennel Dry Extract 5.0%; %, (5) Pigmented dried extract (35.0%), weighed precisely at this compounding ratio and uniformly mixed to obtain a K2 combination, which was used as a sample in the following experimental examples.

Reference example  1. Preparation of experimental animals

1-1. Ready

Sprague Dawley female rats (220-250 g, 8 weeks old) were used in this study in Sam Taco Co., Ltd. (Gyeonggi Province, Korea). The experimental animals were adapted for 7 days in an animal room and used as a control group and an experimental group. The animals were housed in a polycarbonate mice cage at a temperature of 23 ± 3 ° C and a humidity of 50 ± 10% and maintained at a constant 12-hour cycle. The animals were housed in a cage (polycarbonate mice cage) received.

1-2. Establish an animal model

Female white rats were anesthetized by intraperitoneal injection of anesthetic (pentobarbital sodium, 50 mg / kg, Sigma, U.S.A) and incised 1 cm in the median line of the middle part of the spinal cord. The ovaries were exposed and the vessels were ligated with sutures and ovariectomized.

1-3. Drug treatment

Samples of the examples were orally administered at 200, 400 mg / kg for two weeks, 30 minutes before stress application, at the individual compounding ratios of 23.75, 12.5, 5, 23.75,

1-4. Stress imposed

The chronic stress model introduces the most commonly used immobilization stress. Floating stress is caused by floating stress in a certain time (10:00 am) every day using a disposable scalloped plastic bag.

The experimental group consisted of three groups: (I) normal saline, (II) saline + stress + ovariectomy group, (III) drug treatment + stress + drug-treated group do.

Reference example  2. Statistical processing

All the results were statistically analyzed using one way analysis of variance (ANOVA). Significance was tested at P <0.05 level using Turkey's post hoc when significance was recognized.

Experimental Example  1. Identification of antidepressant activity by forced swimming

In order to measure the antidepressant activity of the combination extract according to the forced swimming method, the following test was conducted using the standardized test method FST (RD Porsolt, et al. Behavioral despair in mice: a primary screening test for antidepressants, Arch Int Pharmacodyn Ther, 229, pp. 327-336, 1977).

Using a transparent cylindrical water tank with a depth of 50 cm and a diameter of 30 cm, the rat was put into the water tank for 15 minutes on the first day, and after 24 hours of this test (performed on the second day), climbing, swimming ), And immobility, respectively. At this time, immobility is used as an index of depressive behavior. After 15 minutes of forced swimming, the first drug is administered and the drug is administered 5 hours and 1 hour before the test. After 24 hours, forced swimming in the same environment for 5 minutes, where Climbing, Swimming, and Immobility are measured. A typical immobility is a state in which the mouse is exposed to the surface of the upper body, including the face, while maintaining body balance and no movement of the limbs. Swimming, on the other hand, is a state in which mice move around on the surface of the water and sometimes dive under water. Climbing is the most intense exercise, with the forehead being actively used to move the limb over the acrylic cylinder. To reduce errors and errors caused by human errors, we measured the data with a video camera. Each group consisted of 5-7 animals.

EXAMPLES To determine the antidepressant activity of the samples, the immobility time was measured using FST.

As shown in FIG. 1, the immobility time was 121.0 ± 21.0 sec in the control group, 80.0 ± 13.0 sec in the 200 mg / kg group, and 45.0 ± 15 sec in the 400 mg / kg group, ( P < 0.05).

Experimental Example  2. Tail Suspension Experiment  Confirming antidepressant activity by

(Steru et al., The tail suspension test: a new method for screening antidepressants in mice. Psychopharmacology 85, pp.367 -370, 1985).

2.3.2 Tail suspension test (TST) - Learned helplessness measurement (depression behavior measurement)

Width x Length (50 x 50 cm) Head in the downward direction with a regular hexagonal barrel and fix the tail to the top of the barrel. At this time, let the head fall about 5cm from the floor, but not touch the floor. The experiment is carried out for a total of 6 minutes and immobility is measured for the last 5 minutes.

The antidepressant activity by TST was measured by Steru et al. Tailed mice showed alternating activity and immobility. Anti-depressant drugs decreased the time to show a dose-dependent immobility status, so they measured this time to confirm antidepressant activity. The self-produced TST is used to measure the antidepressant effect after drug withdrawal. In this experiment, the anti-depressive effect was measured by measuring the immobility duration for 10 minutes after attaching a tape 1 cm from the tail tip of the mouse to the edge of the table having a height of 50 cm. The samples were administered at a dose of 200 mg / kg and 400 mg / kg. Each group consisted of 10 animals. TST was used to confirm the antidepressant effect of the sample.

As shown in FIG. 2, the floating time was 115.0 ± 15.0 sec in the control group, 22.0 ± 11.2 sec in the 200 mg / kg group, and 24.0 ± 12.1 sec in the 400 mg / kg group I could. ( P < 0.05).

Experimental Example  3. ELISA analysis

To analyze the immunohistochemical changes of rats after the behavioral tests of the drug extracts of the above examples, ELISA analysis was used as follows. After the behavioral studies in normal animal or depressed animal models, blood is collected from the heart and serum is separated using a high-speed centrifuge (Avanti30, Beckman centrifuge) and stored at -20 ° C. The following day, the concentration of oxytocin in the serum was measured using an ELISA kit (ELISA development system, Cayman, San Antonio, TX, USA).

As a result of the ELISA test,

Results of serum oxytocin measurement

As shown in FIG. 3, serum oxytocin was increased to 216.9 ± 12.0 in the normal group, 189.0 ± 11.0 in the control group, 212.3 ± 14.0 in the 200 mg / kg group and 254.3 ± 12.0 ml / L in the 400 mg / kg group .

serum corticosterone measurement  result

As shown in FIG. 4, the serum corticosterone level was 121.0 ± 11.0% in the control group, 2115.0 ± 12 in the 200 mg / kg group, and 104.3 ± 11.0% in the 400 mg / kg group there was.

Results of serum estradiol measurements

As shown in FIG. 5, the serum estradiol tended to increase to 42.0 ± 12.0% in the control group, 49.0 ± 14 in the 200 mg / kg group, and 58.3 ± 15.0% in the 400 mg / kg group in comparison with the normal group.

Experimental Example  4. Acute toxicity test

An acute toxicity test was conducted by oral administration of the K2 sample obtained in the above Example using the mouse of Reference Example 1. As a result of the acute toxicity test, no case of death was observed at the dose of 2 g / kg of K2 sample extract, and no significant abnormality was found in weight gain, feed intake and the like. Therefore, it was found that the K2 sample extract was a safe drug.

Hereinafter, formulation examples of the composition comprising the K2 sample extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically described.

Formulation example  One. Sanje  Produce

K2 sample ------------------------------------------------ 20 mg

Lactose ------------------------------------------------- - 100 mg

Talc ------------------------------------------------- - 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

Formulation example  2. Preparation of tablets

K2 sample ------------------------------------------------ 10 mg

Corn starch -------------------------------------------- 100 mg

Lactose ------------------------------------------------- - 100 mg

Magnesium stearate ------------------------------------- 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

Formulation example  3. Preparation of capsules

K2 sample ------------------------------------------------ 10 mg

Crystalline cellulose --------------------------------------- 3 mg

Lactose --------------------------------------------- 14.8 mg

Magnesium stearate 0.2 mg &lt; RTI ID = 0.0 &gt;

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

Formulation example  4. Preparation of injections

K2 sample ------------------------------------------------ 10 mg

Mannitol ------------------------------------------------ 180 mg

Sterile sterile distilled water used - 297 mg

Na ₂ HPO ₄ , 12H ₂ O ------------------------------------------ - 26 mg

(2 ml) per 1 ampoule according to the usual injection preparation method.

Formulation Example 5. Preparation of a liquid preparation

K2 sample ------------------------------------------------ 20 mg

Isseong Party ------------------------------------------------ 10 g

Mannitol ------------------------------------------------- - 5 g

Purified water ------------------------------------------------- - Suitable amount

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Formulation example  6. Of health food  Produce

K2 Samples --------------------------------------------- 1000 mg

Vitamin mixture -------------------------------------------

Vitamin A Acetate ----------------------------------- 70 [mu] g

Vitamin E --------------------------------------------- 1.0 mg

Vitamin B1 ------------------------------------------- 0.13 mg

Vitamin B2 - 0.15 mg

Vitamin B6 -------------------------------------------- 0.5 mg

Vitamin B12 ------------------------------------------- 0.2 g

Vitamin C ---------------------------------------------- 10 mg

Biotin ------------------------------------------------ 10 G

Nicotinic acid amide 1.7 mg

Folic acid ------------------------------------------------- - 50 [mu] g

Calcium pantothenate ---------------------------------------- 0.5 mg

Inorganic mixture --------------------------------------------------------------------------

Ferrous sulfate 1.75 mg &lt; RTI ID = 0.0 &gt;

Zinc oxide - 0.82 mg

Magnesium carbonate ---------------------------------------- 25.3 mg

Potassium phosphate monohydrate 15 mg

Secondary Calcium 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 mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Formulation example  7. Manufacture of health drinks

K2 sample ---------------------------------------- 1000 mg

Citric acid ----------------------------------------- 1000 mg

Oligosaccharides ----------------------------------------- 100 g

Plum concentrate ----------------------------------------- 2 g

Taurine --------------------------------------------- 1 g

Add purified water - 900 ml total

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated for about 1 hour at 85 DEG C with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, &Lt; / RTI &gt;

Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.

Claims (7)

(W / w) of a combination herbal medicine comprising, as an active ingredient, a water extract of a combination herb medicine formulated in an amount of 1 to 10 parts by weight, a licorice 1 to 10, a fennel 1 to 10, a soybean 1 to 10 and a pomegranate 1 to 20 parts by weight A pharmaceutical composition for prevention and treatment. delete delete (W / w) of a combination herbal medicine comprising, as an active ingredient, a water extract of a combination herb medicine formulated in an amount of 1 to 10 parts by weight, a licorice 1 to 10, a fennel 1 to 10, a soybean 1 to 10 and a pomegranate 1 to 20 parts by weight Health functional foods for prevention and improvement. (W / w) of a combination herbal medicine comprising, as an active ingredient, a water extract of a combination herb medicine formulated in an amount of 1 to 10 parts by weight, a licorice 1 to 10, a fennel 1 to 10, a soybean 1 to 10 and a pomegranate 1 to 20 parts by weight Prevention and improvement health supplements. delete delete

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