KR20110073653A - Pharmaceutical composition for preventing or treating anxiety syndrome comprising 4-o-methylhonokiol as active ingredient - Google Patents

Abstract

PURPOSE: A medicinal composition containing 4-O-methylhonokiol is provided to prevent and treat cranial nerve system-associated anxiety. CONSTITUTION: A medicinal composition for preventing and treating cranial nerve system-associated anxiety contains 4-O-methylhonokol of chemical formula 1 or pharmaceutically acceptable salt thereof as an active ingredient. The medicinal composition further contains pharmaceutically acceptable carrier, diluent, or excipient.

Description

Pharmaceutical composition for preventing or treating anxiety syndrome comprising 4-O-methylhonokiol as active ingredient

The present invention relates to a pharmaceutical composition for anti-anxiety prevention and treatment containing 4-O-methylhonokiol (4-O-methylhonokiol) as an active ingredient, more specifically 4-O-methylhonokiol or a pharmaceutical thereof The present invention relates to a pharmaceutical composition or a health functional food for preventing and treating anxiety caused by abnormalities of various cranial nervous systems containing a scientifically acceptable salt as an active ingredient.

As the economic growth rate has recently declined, more and more people are feeling depressed or anxious, and the number of patients with depression or anxiety is increasing in relation to rapidly changing family relationships such as divorce and child problems. Anxiety is not only a widespread very unpleasant or vague feeling of anxiety, but also physical symptoms associated with it, accompanied by symptoms such as palpitations, sweating, and behavioral symptoms such as irritability and pacing (Robert F. Scmidt., Human Physiology , pp 366; Min Sung Gil, Modern Psychiatry , pp238-40; Argyropoulos SV et al., Pharmacol. Ther. 88 , pp213-27, 2000). Increasingly, patients are receiving specialized psychiatric counseling at hospitals for depression or anxiety, and anti-anxiety sales increased by 200 percent in 2002. It is no exaggeration to say that depression or anxiety can account for most of the mental illnesses of modern people, and many people suffer from it.

Anxiety is a type of defense system to protect the human body. Excite the autonomic nervous system, and when the anxiety is too frequent, fatigue or excessive activation of the sympathetic nerve due to stress and overwork, it causes anxiety in life and develops anxiety disorder. If anxiety persists or worsens, it develops into anxiety disorders. Anxiety disorders include panic disorder, obsessive compulsive disorder, stress disorder, social phobia, and panic anxiety disorder. In particular, social phobia and panic anxiety disorders are suffering from many people even though they are not well known.

In the US, 13% of the population has social phobia, and 2% of the population in Korea have severe social phobia who needs treatment.At least 20% of the population suffers from social phobia. It is assumed that there is. Symptoms of panic anxiety disorder can be mistaken for cardiovascular disease, hyper-insulinemia, hyperthyroidism, etc., which can increase the cost of examination for diagnosis.

Therefore, in the United States, it has been reported that the social cost of panic anxiety disorder, including the cost of these tests, reaches $ 26 billion (3.1 billion won) a year, and panic anxiety disorder often requires more than one year of drug treatment. There is a need to reduce the pain caused by anxiety disorders and to improve the quality of human life, and to reduce the social or personal costs caused by anxiety disorders.

When there is anxiety, the entire brain enters an arousal state, thus disrupting peripheral behavior, autonomic nervous system, sensations, and perception. The main organs involved are the cerebral limbic system (especially the hippocampus and the subject society), the cerebral cortex (frontal lobe, temporal lobe), hypothalamus, ascending reticular body and pituitary gland. Recent brain imaging studies have shown that anxiety is associated with disorders in the right hemisphere and other frontal, temporal, and occipital lobe disorders (Robert F. Scmidt., Human Physiology , pp366; Min Sung-gil, Modern Psychiatry , pp238-40). .

The ideal anti-anxiety drug will not result in excessive drowsiness during the day, cause physical and mental dependence, and calm the patient.

Representative anti-anxiety agents currently available include benzodiazepine-based drugs, including diazepam, oxazepam, prazepam, lorazzepam, alprazolam, and helazepam ( helazepam) and clonazepam; these drugs are also used primarily for the purpose of sedation and sleep (Yoon Do Joon, Side Effects of Psychiatric Drugs, Journal of the Korean Medical Association , 38 (10), pp1196-1202, 1995) .

Benzodiazepine-based medications are most commonly by using anti-anxiety, mechanism of action of this drug is typically an inhibitory neurotransmitter, gamma in the central nervous system - open more often the passage Cl ^- adjacent to increase the affinity of the amino acid (GABA) receptor-Cl ^- It has been found to increase the permeability of ions. These benzodiazepines are effective immediately, but they are addictive and addictive.If you don't use the medicine according to the doctor's treatment, symptoms may recur or withdraw, and other side effects include drowsiness, ataxia, orthostatic hypotension, respiratory depression, headache , Chronic sleep disorders, and liver disease have been reported (Mary J. Mycek, et al., Pharmacology 2nd edition , Lipincott Williams & Wilkins, pp89-93, 2000). The development of new structures of drugs is required.

Oriental medicine recognizes depression, anxiety, and nervousness as being caused by fever, blood, heart, heart, and nasal congestion. It is known that depression, anxiety, insomnia, dreams, fears, cramps, frenzy, falsification. In addition, symptoms such as major (臟 躁), air force (氣 逆), dividers (경계), alert (驚悸) is mainly classified as a panic disorder. Significant mental disorders in response to intense stress, also called giran, are included in the category of depression or anxiety in modern medicine.

These mental nervous system diseases are mainly treated with a drug group called anxin drug (安神 藥物), which reduces depression or anxiety, and has a sedative hypnotic and anticonvulsive effect. About this treatment, herbal medicine says that it can be cured by using mental and physical stabilization in case of surprise, fear, depression or anxiety. Drugs belonging to Ahnsin are known to have the effect of nerve stabilization, although they have their own characteristics (Songbon polar acid, synergistic consideration of neurosis, Oriental Medicine , pp65-8, 1986) ).

Numerous neuropharmacological studies have been reported on drugs with antagonism . In particular, Sanjoin ( Zyziphus jujuba Mill., 酸棗仁 ) is known to have anti-anxiety effects at low doses and to be sedative at high doses (Wu SX, et al. , China Journal of Chinese Materia Medica ., 18 , pp685-704), In addition to the Anxin drug group, Gastrodia elata Blume. (天麻) has been shown to reduce spontaneous motility and pentobarbital induced in mouse studies. It is known to have sedative effects such as prolonged sleep (Huang JH, Yi Xue Ke Xue Yuan Xue Bao. , 11 , pp147-50, 1989).

Thick leaf has been used as a health agent for thousands of years in Korea, Japan, and China, and leaves have been eaten for food in Korea and Japan. Japanese humbac (M. obovata THUNB.) Contains 1% of essential oils. The essential oils include beta-eudesmol and beta-pinene (β). -pinene is known and contains phenolic magnolol, honokiol, etc. (Information Sup, Shin Mingyo, Pharmacological Dictionary, Younglimsa, pp469-471, 1998)

The pharmacological effects of 4-O-methyl-honochiol of such thick foil have already been disclosed in various patent documents, and the treatment of amyloid-related diseases containing '4-0-methyl-honochiol of Korea Patent Registration No. 10-926466 Alternatively, the prophylactic pharmaceutical composition discloses that 4-O-methylhonokiol is effective for treating or preventing amyloid-related diseases such as Alzheimer's disease, cognitive impairment, memory loss and amyloidosis.

On the other hand, the Republic of Korea Patent Publication No. 10-2008-105526 'anti-inflammatory, anti-allergic and anti-wrinkle composition containing 4-O-methyl honokiol as an active ingredient in the thick extract or separated therefrom 4-O-methyl hono It is disclosed that the chiol can be used for external application of the skin for anti-inflammatory, anti-allergic and wrinkle improvement by showing antihistamine activity enhancement, elastase inhibitory effect, and the like.

However, the anti-anxiety preventive and therapeutic effects of 4-O-methylhonokiol have not been disclosed in the literature so far. Accordingly, the present inventors measured the anti-anxiety effect of 4-O-methylhonokiol by using an elevated plus-maze model, and the degree of anti-anxiety effect shown at this time was diazepam, a drug used as an existing anti-anxiety agent. As a control drug, the behavioral pharmacological comparison result showed that 4-O-methylhonokiol showed more than equivalent activity compared to diazepam, and also confirmed that it did not show the sleep effect of the experimental animal which is a side effect of diazepam. The present invention has been completed.

The problem to be solved by the present invention is to measure the anti-anxiety effect of 4-O-methyl honokiol using an elevated plus-maze model. In addition, diazepam, a drug used as an anti-anxiety drug, was used as a control drug to compare behavioral pharmacology.

An object of the present invention is to provide a pharmaceutical composition for the prevention and treatment of cerebral nervous system-related anxiety, containing 4-O-methylhonokiol represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

Formula 1

In another aspect, the present invention is characterized in that the active ingredient contains 4-0-methylhonokiol by 0.001 ~ 90% by weight relative to the total weight of the composition, and comprises 4-0-methylhonokoiol and a pharmaceutically acceptable carrier, diluent or excipient do.

In addition, the cerebral nervous system-related anxiety is characterized by including panic disorder, obsessive compulsive disorder, stress disorder, social phobia, panic anxiety disorder.

It is another object of the present invention to provide a health functional food for the prevention and treatment of cerebral nervous system-related anxiety, which contains 4-0-methylhonokiol, which is isolated from grobac extract or manufactured synthetically as a functional ingredient.

At this time, the health functional food is characterized in that the form of pills, powder, granules, acupuncture, tablets, capsules or beverages.

The effect of the present invention is that the composition containing the 4-O-methyl honokiol compound as an active ingredient exhibits anti-anxiety pharmacological effect, the composition containing the 4-O-methyl honokiol compound due to various brain nervous system abnormalities It is to provide a pharmaceutical composition and dietary supplement for the prevention and treatment of anxiety.

The present invention is described in more detail below.

4-O-methyl honokiol used in the present invention was obtained from Bioland Co., Ltd., and the separation method of 4-O-methyl honokiol from the thick extract is disclosed in Korean Patent Application Publication No. 10-2009-94916. It was isolated by the method disclosed in the following "Hair extract or composition for promoting hair growth or containing 4-O-methyl-honokiol separated therefrom".

The present invention provides a pharmaceutical composition for the prevention and treatment of cerebral nervous system-related anxiety, which contains 4-O-methylhonokiol compound, which is a compound isolated from Japanese hawthorn leaf extract, as an active ingredient.

In addition, Japanese hummus leaves have been used for a long time as herbal medicines and edible extracts of the present invention or compounds isolated therefrom also have no problems such as toxicity and side effects.

Compounds having a prophylactic and therapeutic effect on cerebral nervous system-related anxiety of the present invention can be synthesized through synthesis and fractionation of conventional substituents. (Herbert O. House: Modern Synthetic Reactions , 2nd Ed ., The Benjamin / Cummings Publishing Co., 1972).

The compounds of the present invention can be prepared with pharmaceutically acceptable salts and solvates according to methods conventional in the art.

As the pharmaceutically acceptable salt, acid addition salts formed by free acid are useful. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equal molar amounts of the compound and acid or alcohol (eg, glycol monomethyl ether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

In this case, organic acids and inorganic acids may be used as the free acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. may be used as the inorganic acid, and methanesulfonic acid, p -toluenesulfonic acid, acetic acid, trifluoroacetic acid, citric acid may be used as the organic acid. , Maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid, and the like.

Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salts, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Pharmaceutically acceptable salts of the compounds of the invention include salts of acidic or basic groups which may be present in the compounds of the invention, unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, and methods or processes for preparing salts known in the art It can be prepared through.

In order to examine the anti-anxiety effect of the compound of the present invention obtained by the above method, the 4-O-methyl honokiol of the present invention is anti-anxiety using an levated plus-maze model and a hole board In comparison with the behavioral pharmacological study using diazepam, the existing drug, 4-O-methylhonokiol showed a similar effect at a lower concentration than diazepam. It was confirmed that the sleep effect is not shown.

The pharmaceutical composition having a prophylactic and therapeutic effect of cerebral nervous system-related anxiety of the present invention comprises 0.001 to 90% by weight of the compound, and preferably 0.1 to 50% by weight, based on the total weight of the composition.

Pharmaceutical compositions comprising the compounds of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The pharmaceutical compositions comprising the compounds according to the present invention may be prepared in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used.

Carriers, excipients and diluents that may be included in the composition comprising the compound include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, or the like. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

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

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried 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, glycerogelatin and the like can be used.

Preferred dosages of the compounds of the present invention depend on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention is preferably administered at 0.0001 to 100 mg / kg body weight, preferably 0.01 to 10 mg / kg body weight per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The compounds of the present invention can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intra-cerebroventricular injection.

The present invention provides a dietary supplement for the prevention of cerebral nervous system-related anxiety comprising the 4-O-methyl-honokinool compound and a food acceptable additive.

The composition containing the 4-O-methylhonokiol compound of the present invention may be used in a variety of drugs, foods and beverages for the prevention of cerebral nervous system-related anxiety. Examples of the food to which the 4-O-methylhonokiol compound of the present invention can be added include various foods, for example, beverages, gums, teas, vitamin complexes, health supplements, and the like. , Tablets, capsules or beverages.

In this case, the amount of the 4-O-methylhonokiol compound in the food or beverage may generally be added in an amount of 0.001 to 30% by weight of the total food weight of the health food composition of the present invention, and preferably 0.01 to 15% by weight. . In the case of a health beverage composition it may be added in a ratio of 0.02 to 10 g, preferably 0.3 to 1 g based on 100 ml.

Food supplement additives as defined herein include food additives customary in the art, such as flavorings, flavoring agents, colorants, fillers, stabilizers, and the like, and are exemplified below.

The health beverage composition of the present invention has no particular limitation on the liquid component except for containing the extract as an essential ingredient in the indicated ratio, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; Polysaccharides such as dextrin, cyclodextrin; Conventional sugars such as and the like and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), 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, and the like. The compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is usually selected in the range of 0 to 20 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention will be described in more detail with reference to the following examples. However, these examples do not limit the scope of the present invention.

Example 1 Anti-Anxiety Effect of 4-O-Methyl Honokiol by Elevated Plus-Maze Laboratory Animal Model

5-6 week old ICR male mice (20-28 g) (Korea Biolink) were raised in an animal room maintaining moderate temperature (22 ± 2 ° C) temperature and circadian (12 hours), and 10 animals were group 1 The cage was freely supplied with water and food. Experimental mice were used for 1 week prior to experiment.

In order to investigate the anti-anxiety effect of 4-O-methylhonokiool, the experiment was performed by applying the elevated plus-maze experimental animal model described in the existing literature (Dawson, GR et al., Trends in Pharmarcological). Sciences, 16 , pp 33-36, 1995).

The plus-maize device has a central plane (5 x 5 cm) crosswise at a height of 38 cm from the bottom, with two open arms (30 x 5 x 15 cm) and two closed arms connected to it. , 30 x 5 x 15 cm). The planar layers of each branch are made of wood, and the two closed arms have a protective wall made of transparent material to keep the animals from falling.

Elevated Plus-Maze model experiments were conducted in a dim room in a dim room, and the experimental rats were placed on the plus-mages one hour after a single dose of sample, and the number of open arms entered and stayed for five minutes was measured. Plus-Maze was dehumidified and wiped with a dry towel every time it was measured. All experiments were recorded with a video camera mounted on plus-maze. The activity against open cancer was expressed as a percentage of 1) total residence time in open arm / plus-maze and 2) entry and exit number in open and closed arm (100 x open / total entries). Each arm was considered to have entered four feet and two feet to come out. Behavioral experiments were conducted between 13: 30-16: 30 in quiet, low light (50 lux).

As a result of the experiment, as shown in Figures 1a and 1c, the experimental rats administered diazepam (2 mg / kg), a positive control drug increased the percentage of open cancer access, 4-O-methyl honokiol (0.1 , 0.25, 0.5 mg / kg), like diazepam, it was confirmed that the percentage increase in the number of entry and exit of open cancer in a concentration-dependent manner. In addition, as shown in Figures 1b and 1d, diazepam (2 mg / kg) showed a tendency to increase the retention time percentage of open cancer, 4-O-methyl honokiol (0.1, 0.25, 0.5 mg / kg) In addition, the percentage of retention time of open cancer increased concentration-dependently. It was confirmed that 4-O-methyl honokiol showed a strong anti-anxiety effect by confirming that the percentage of entry and residence time of open cancer increased at a lower dose than diazepam, a positive control drug. .

The experimental results were expressed as mean error, and the significance of the compound effect was evaluated by the Variance Analysis (ANOVA). p0.05, p0.01, p0.005 are shown in comparison with the control.

Example 2 Expression Changes of GABA _A Receptor Subunits by 4-O-Methyl Honokiol in Elevated Plus-Maze Laboratory Animal Models

Mouse brains in lysis buffer [50 mM Tris pH 8.0, 150 mM NaCl, 0.02% sodium azide, 0.2% SDS, 1 mM PMFS, 10 μl / ml aprotinin, 1% igapel 630 ( Sigma Chem. Co. St. Louis, MO, USA), 10 mM NaF, 0.5 mM EDTA, 0.1 mM EGTA, and 0.5% sodium deoxycholate], and centrifuged at 23,000 g for 15 minutes. It was. Equal amounts of protein (50 μg) were separated on SDS / 12% -polyacrylamide gels and then transferred to nitrocellulose membranes (Hybond ECL, Amersham Pharmacia Biotech Inc., Piscataway, NJ, USA). The blot was blocked for 2 hours at room temperature with 5% (w / v) dry skim milk dissolved in Trisbuffered saline [10 mM Tris (pH 8.0) and 150 mM NaCl] containing 0.05% Tween-20. . Membranes were incubated for 5 hours at room temperature with the following specific antibodies: mouse polyclonal antibodies for α-subunit and β-subunit (1: 500 dilution; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). The blot was then incubated with the corresponding conjugated anti-rabbit immunoglobulin G-horse radish peroxidase (1: 4,000 dilution, Santa Cruz Biotechnology Inc.). Immunoreactive proteins were measured with an ECL western blotting detection system.

As shown in FIG. 1A, the expression of GABA _A receptor subunits a-subunit and b-subunit in the cerebral cortex and hippocampus of rats administered with the positive control diazepam (2 mg / kg) was shown in FIG. 4-O-methylhonokiol (0.5 mg / kg), like diazepam, increased the expression of α-subunit and β-subunit. The relative density of protein bands was scanned with a MyImage (SLB) densitometry and quantified by Labworks 4.0 software (UVP Inc.). The graph was calculated with α-subunit / β-actin and β-subunit / β-actin.

Example 3 Anti-anxiety Effect of 4-O-Methyl Honokiol by Head Dips Experimental Animal Model

The Hall board test actively connects to the novelty and is used as an indicator of anti-anxiety effect. When an animal is placed in a box cage with four holes, it is frightened as a probe and looks into the hole.

As a result of the experiment, as shown in Figure 3, the experimental rats administered the positive control diazepam (2 mg / kg) significantly increased the number of head dip compared to the control group, 4-O-methyl honokiol (0.1, 0.25, 0.5 mg / kg), like diazepam, it was confirmed that 4-O-methyl-honokiol showed a strong anti-anxiety effect by increasing the number of head dips in a concentration-dependent manner.

The experimental results were expressed as mean error, and the significance of the compound effect was evaluated by the Variance Analysis (ANOVA). p0.05, p0.01, p0.005 are shown in comparison with the control.

Example 4 Cl ⁻ Influence of 4-O-Methyl Honokiol in Cortical Neurons

Cultured cortical neurons from the brains of rats are isolated and measured for Cl ^- concentration in resting cells using standard Cl ^- solution. As shown in FIG. 4, Cl ⁻ concentrations in resting cells were increased in the cortical neurons treated with the positive control diazepam (5 mM) as compared to the control group, and 4-O-methylhonokiol (2 , 5, 10 mM), like diazepam, was confirmed that 4-O-methylhonokiol exhibited a strong anti-anxiety effect by increasing the concentration of Cl ^- concentration in resting cells compared to the control group.

The experimental results were expressed as mean error, and the significance of the compound effect was evaluated by the Variance Analysis (ANOVA). p0.05, p0.01, p0.005 are shown in comparison with the control.

The preparation example of the pharmaceutical composition containing 4-O-methyl-honokinool of the present invention will be described, but the present invention is not intended to be limited thereto but merely to explain in detail.

Formulation Example 1 Preparation of Powder

4-O-methylhonokiol compound 300 mg

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

4-O-methylhonokiol compound 50 mg

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

4-O-methylhonokiol compound 50 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

4-O-methylhonokiol compound 50 mg

Appropriate sterile distilled water for injection

pH adjuster

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

4-O-methylhonokiol compound 100 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method for preparing a liquid, each component is added and dissolved in purified water, lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by adding purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Food

4-O-methylhonokiol compound 1000 mg

Vitamin mixture proper amount

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

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic 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 mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 7 Preparation of Healthy Drink

4-O-methylhonokiol compound 1000 mg

Citric acid 1000 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 a conventional healthy beverage production method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained by sterilization in a sterilized 2 L container, sealed sterilized and then stored in the present invention For the preparation of healthy beverage compositions.

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Figure 1a is a diagram showing the number of entry and exit in the open arm by the concentration of diazepam and 4-O-methyl honokiol for open arm entries in the elevated plus-maze (*), indicating the degree of significance p <0.05, ** P <0.01 are shown compared to the control.

Figure 1b is a diagram showing the retention time of the hourly open cancer of diazepam and 4-O-methyl-honokiol for open arm entries in the elevated plus-maze, indicating the degree of significance * p <0.05, ** P <0.01 were obtained compared to the control.

Figure 1c is a diagram showing the number of access to open cancer by observing the effects of diazepam and 4-O-methylhonokiol on open arm entries in the elevated plus-maze for one week, * P <0.05 and ** P <0.01 indicating the degree of significance were shown in comparison with the control.

FIG. 1D is a diagram showing the retention time in open cancer by observing the effects of diazepam and 4-O-methylhonokiol on open arm entries for one week in elevated plus-maze. * P <0.05 and ** P <0.01 indicating the degree are shown in comparison with the control.

2 is a tissue of the mouse brain showing the effect of diazepam and 4-O-methylhonokiol on open arm entries in elevated plus-maze through Western blot. It is a diagram showing the change in the expression of GABA alpha and GABA beta, and the degree of expression of each protein is graphed, * p <0.05, ** P <0.01 indicating the degree of significance is shown compared to the control.

FIG. 3 is a diagram showing the effect of diazepam and 4-O-methylhonokiol on the number of head dipps in the hole-board test. * P <0.05, ** P <0.01 is shown compared to the control.

Figure 4 is a chloride introduced (Cl ^- influx) and experimental diagram showing the effect of diazepam and a 4-O- methyl hono kiol for the concentration of chloride in the inflow cells, * p <0.05 indicating a significant degree, ** P <0.01 Is shown in comparison with the control.

Claims (5)

Pharmaceutical composition for the prevention and treatment of cerebral nervous system-related anxiety containing 4-O-methyl-honokiol represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient Formula 1

The brain according to claim 1, wherein the active ingredient contains 0.001-90% by weight of 4-0-methylhonokiol, based on the total weight of the composition, and 4-0-methylhonokiol and a pharmaceutically acceptable carrier, diluent or excipient. Pharmaceutical composition for the prevention and treatment of nervous system related anxiety According to claim 1, CNS-related anxiety, pharmaceutical composition for the prevention and treatment of cerebral nervous system-related anxiety, including panic disorder, obsessive compulsive disorder, stress disorder, social phobia, panic anxiety disorder, etc. Health functional food for the prevention and treatment of cerebral nervous system related anxiety, containing 4-0-methylhonokiol isolated from synthetic extract or synthetically prepared as functional ingredient 5. The health functional food of claim 4, wherein the health functional food is in the form of pills, powder, granules, acupuncture, tablets, capsules or beverages.

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