KR20100092824A - Pharmaceutical composition comprising chlorogenic acid or its salt as effective ingredient for preventing or treating of learning or memory disorder, or dementia - Google Patents

Abstract

PURPOSE: A composition containing chlorogenic acid is provided to use as an agent for preventing learning disorder, memory disorder, and dementia disorder. CONSTITUTION: A pharmaceutical composition for preventing or treating learning disorder or memory disorder contains pharmaceutically effective amount of chlorogenic acid or pharmaceutically acceptable salt and pharmaceutically acceptable carrier. A pharmaceutical composition for preventing or treating dementia contains pharmaceutically effective amount of chlorogenic acid or pharmaceutically acceptable salt and pharmaceutically acceptable carrier.

Description

Pharmaceutical composition comprising chlorogenic acid or its salt as effective ingredient for preventing or treating of learning or memory disorder, or dementia}

The present invention relates to a pharmaceutical composition for preventing or treating learning disorders, memory disorders or dementia, and chlorogenic acid or a pharmaceutically acceptable salt thereof as an active ingredient, and a food or health functional food for learning or memory enhancement. .

Memory refers to the process of acquiring new information, learned experiences, and knowledge from the environment, encoding it in specific parts of the brain, storing it, and recalling it (William F Ganong; Ganong's Physiology, Seoul, Hanwoori, p289, p291-). 292, 1999). The process of memory is divided into several stages: acquisition, encoding, reinforcement, retention, and recall.

Modern society is a complex and specialized trend, and accordingly, a large amount of information and learning amount is required, and therefore effective brain activity is required. Many people are suffering from memory loss due to aging and disease, and high mental activity is required as the life span of human beings is extended. In order to cope with the situation appropriately, various efforts have been made to clear the brain, improve memory, and require mental activities, and the development of medicines, health functional foods, etc., which can effectively promote such functions, It is required.

Dementia is divided into Alzheimer's dementia and vascular dementia. Alzheimer's dementia is a type of dementia commonly observed in the West, and is a disease in which amyloid beta accumulates in certain parts of the brain, resulting in destruction of cholinergic neurons. In contrast, vascular dementia is a dementia caused by hypertension, stroke, and hyperlipidemia due to abnormal blood vessels leading to the brain, resulting in the death of nerves in the hippocampus and limbic system. There is still little research on vascular dementia.

Although the amyloid hypothesis was introduced in 1991 that amyloid beta deposition is a cause of Alzheimer's dementia, no substance has been developed to treat dementia.

Clinically used drugs approved by the FDA as a treatment for dementia are acetylcholinesterase inhibitors (tacrine tacrine, donepezil donepezil, galantamine galanthamine, rivastigmine rivastigmine) or NMDA receptor antagonists (memantine) .

Among these, acetylcholine having a quaternary amine structure is a neurotransmitter and acetylcholinesterase hydrolyzes acetylcholine into choline. In patients with dementia, the concentration of acetylcholine, a neurotransmitter, is reduced, and when acetylcholinesterase is inhibited, the concentration of acetylcholine in the brain is increased and symptoms of dementia are improved.

When oxidative stress is applied in vivo, the oxygen in the living body is superoxide radical (O ^2- ), hydroxyl radical (OH-), hydrogen peroxide (H ₂ O ₂ ), singlet It converts into highly reactive active oxygen, such as singlet oxygen (1O ₂ ). Such active oxygen causes deadly oxygen toxicity in the living body and induces non-selective and irreversible destruction of cell components such as lipids, proteins, sugars, DNA, etc., aging, brain diseases such as stroke, Parkinson's disease, It is known to cause various diseases such as heart disease, ischemia, arteriosclerosis, skin disease, digestive disease, inflammation, rheumatism and autoimmune diseases (Cross, EE, et al., Ann. Intern. Med., 107, 526, 1987; Halliwell, B., FASEB J., 1, 358, 1987; Alderson, RRL, et al., Proc. Natl. Acad. Sci. USA, 85, 2706, 1988). In addition, the active oxygen induces peroxidation of substances in vivo to produce various peroxides, including lipid peroxides. These peroxides cause oxidative destruction of cellular components and cause various functional disorders of the living body, thereby causing aging and disease. Cause was reported (Harmon, D., Alan R Liss, New York., 3-49, 1986). As oxidative stress is found to be an important cause of various diseases including aging, the possibility of inhibiting aging and preventing or treating various diseases mentioned above by using an antioxidant which has the activity of removing free radicals in vivo is known. It is greatly highlighted.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating learning disorders, memory disorders and dementia comprising chlorogenic acid or a pharmaceutically acceptable salt thereof as an active ingredient. In addition, an object of the present invention is to provide a food or health functional food for learning or memory enhancement, comprising chlorogenic acid or a pharmaceutically acceptable salt thereof as an active ingredient.

The present inventors have conducted learning and memory enhancement using animal models, improvement experiments, acetylcholinesterase inhibitory experiments and free radical scavenging experiments. As a result, chlorogenic acid enhances and improves memory, and inhibits acetylcholinesterase effects. And the present invention was completed by confirming the ability to eliminate free radicals.

According to one aspect of the present invention, a pharmaceutical composition and a medicament for preventing or treating learning disorders, memory disorders or dementia diseases comprising chlorogenic acid having the formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient Is provided.

화학식 (I)

(I)

3-[[3- (3,4-Dihydroxyphenyl) -1-oxo-2-propenyl] oxy] -1,4,5-trihydroxy cyclohexanecarboxylic acid {3-[[3 -(3,4-Dihydroxyphenyl) -1-oxo-2-propenyl] oxy] -1,4,5-trihydroxycyclohexanecarboxylic acid} or 3-carffeoylquinic acid

According to the present invention, chlorogenic acid exhibited the ability to inhibit acetylcholinesterase, was excellent in the ability to eliminate free radicals, and the results of animal experiments showed an improvement in working memory in the scopolamine-induced forgetfulness model. In addition to the remarkable effects of forgetfulness itself, it can be used as a preventive or therapeutic agent for learning disorders, memory disorders and dementia, and also in the manufacture of foods, dietary supplements or functional foods to enhance learning or memory. Can be used.

According to another aspect of the present invention, there is provided a food, health supplement or health functional food, comprising chlorogenic acid of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

Chlorogenic acid of the present invention exhibited the ability to inhibit acetylcholinesterase, was excellent in the ability to eliminate free radicals, and in addition to the work memory, scopolamine-induced forgetfulness model showed the effect of improving working memory, The improvement effect of forgetfulness itself was also remarkable, and can be used as a preventive or therapeutic agent for learning disorders, memory disorders, and dementia, and can also be used to manufacture health functional foods to enhance learning or memory.

Chlorogenic acid of Formula (I) is a substance contained in higher plants, especially fresh coffee beans and roasted coffee beans, and can be extracted using HPLC-UV as a methanol / water mixed solvent (Bicchi et al. J. Agric.Food Chem., 43, pp 1549-1555 (1995)).

The salts of chlorogenic acid include conventional non-toxic salts, for example hydrochloride, sulfate, maleate, sodium salt, carbonate, tartrate, bitartrate, nitrate ), Benzoate, lysine, citrate, bromide, hydrobromide, and the like.

As used herein, "learning" refers to an ability or behavior that can perceive and change one's own behavior, and includes spatial perception, cognition, concentration, and the like.

"Learning disorder or learning disability" refers to "learning disorder or learning disability" due to a variety of causes, regardless of IQ, for example, depression, anxiety, obsessive compulsive disorder, social and environmental factors (failure, poverty, missing family, stress, etc.). "Ability is lowered than normal, and it includes a decrease in spatial perception, a decrease in cognition, a concentration, and a decrease in academic achievement of children.

The term "memory" or "memory" refers to the ability to acquire new information, learned experience, and knowledge from the environment, encode it in a specific area of the brain, store it, and recall it.

"Memory disorder" or "memory deficiency" refers to a condition in which the "memory" is lowered than a normal person due to various causes such as trauma, attention deficit, aging, disease, and amnesia, concentration disorder, Loss of spatial perception, slowing of learning ability, loss of cognition.

According to one embodiment of the invention, the chlorogenic acid of formula (I) or a pharmaceutically acceptable salt thereof is 0.01 per day to prevent or treat a learning disorder, memory disorder or dementia, or to enhance learning or memory. To the subject may be administered to the subject once or divided into several doses of 500 mg / kg, and preferably may be administered once or divided into several doses of 5 to 100 mg / kg per day, Appropriate doses may be selected by those skilled in the art depending on body weight, extent of disease, drug form, route of administration and duration.

According to the present invention, the pharmaceutical composition for the prevention or treatment of learning disorders, memory disorders or dementia diseases comprising chlorogenic acid of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient is a pharmaceutically acceptable carrier. May be included. The pharmaceutical composition may preferably be used for the prevention or treatment of vascular dementia disease.

Chlorogenic acid or a pharmaceutically acceptable salt thereof may be included in a ratio of 0.001 to 50% by weight based on the total weight of the pharmaceutical composition, but those skilled in the art may appropriately adjust the ratio according to the purpose of use.

The pharmaceutical composition of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles for the purpose of preventing or treating learning disorders, memory disorders or dementia diseases.

The composition according to the present invention may be formulated into a pharmaceutical composition including a pharmaceutically acceptable carrier, oral dosage form such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc. according to conventional methods, It may be formulated in the form of external preparations, suppositories, and sterile injectable solutions.

The pharmaceutically acceptable carrier is 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, mineral oil and the like. Also included are diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and the like. Oral solid preparations include tablets, pills, powders, granules, capsules and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose or lactose. ), Gelatin, and the like, and may include a lubricant such as magnesium stearate, talc, and the like. Oral liquid preparations include suspensions, solvents, emulsions, syrups, and the like, and may include water, diluents such as liquid paraffin, wetting agents, sweeteners, fragrances, preservatives, and the like. Parenteral preparations include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and ethyl. Injectable esters such as oleate and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogelatin and the like can be used.

The food, dietary supplement or dietary supplement of the present invention is 0.01 to 95% by weight of chlorogenic acid or its pharmaceutically acceptable salt, preferably 1 to 80, based on the total weight of the composition for the purpose of learning or memory enhancement. It may be included in the ratio of weight percent, but those skilled in the art can adjust the ratio as appropriate for the purpose of use.

The health functional food of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like for the purpose of learning or memory enhancement.

"Health functional food" means a food manufactured and processed using raw materials or ingredients with functional functions useful to human body under Act No.6727 of the Health Functional Food Act, and controls nutrients on the structure and function of the human body or It means ingestion for the purpose of obtaining a useful effect for health use such as physiological action.

The health functional food of the present invention may include a conventional food additive, and the suitability as the "food additive" is applicable in accordance with the General Regulations of the Food Additives Code and General Test Methods, etc. approved by the Food and Drug Administration, unless otherwise specified. Judge according to the standards and standards for the item.

Examples of the items listed in the "Food Additive Revolution" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamon acid, natural additives such as color pigments, licorice extracts, crystalline cellulose, high color pigments, guar gum, Mixed preparations, such as a sodium L- glutamate preparation, an addition of an alkali, a preservative, and a tar coloring agent, etc. are mentioned.

For example, a dietary supplement may be prepared by granulating a mixture of chlorogenic acid with excipients, binders, disintegrants, and other additives in a conventional manner, and then compressing the mixture with a lubricant or the like. Direct compression molding is possible. In addition, the health functional food in the form of tablets may contain a mating agent and the like, if necessary, may be coated with a suitable coating agent.

Hard capsules of the health functional foods in the form of capsules may be prepared by filling a conventional hard capsule with chlorogenic acid or a mixture of additives such as excipients or granules or peeled granules thereof. A mixture with additives such as excipients may be prepared by filling a capsule base such as gelatin. The soft capsule agent may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, as necessary.

The dietary supplement in cyclic form may be prepared by molding a mixture of chlorogenic acid by excipients, binders, disintegrants, etc. in a suitable manner. You can also give a favor.

The health functional food in the form of granules may be prepared by granulating a mixture of the chlorogenic acid as an excipient, a binder, a disintegrating agent, etc. in a suitable manner, and may contain a flavoring agent, a mating agent and the like as necessary.

The term definitions of the excipients, binders, disintegrants, glidants, copulation agents, flavoring agents, etc. of the present invention are described in documents known in the art and include those having the same or similar functions. Sacrament, Korean College of Pharmacy, 5 revised edition, p33-48, 1989).

Hereinafter, the present invention will be described in more detail through Examples and Test Examples. However, these Examples and Test Examples are for illustrating the present invention, and the scope of the present invention is not limited to these Examples and Test Examples.

[Example]

Example 1 Drugs and Reagents

Chlorogenic acid, scopolamine, acetylthiocholine iodide and DTN were purchased from Sigma-Aldrich Chemistry Co. and used.

Example 2 Preparation of Experimental Animals

6 weeks old ICR male mouse (25-30 g) It was supplied from (Pyeongtaek, South Korea) and used in the animal breeding room of Sungkyunkwan University Pharmacy for more than a week and adapted to it. The water and feed were freely consumed, and the temperature (23 ± 2 ℃) and humidity (55 ± 10%) were used. And the contrast cycle (12 hours) were to be adjusted automatically.

Example 3. Statistical Processing

All experimental results were statistically analyzed using ANOVA (one and two way analysis of variance). When significance was verified, significance test was performed at p <0.05 level using Newman-Keul's and Dunnett test.

Example 4 Inhibitory Effect of Chlorogenic Acid on Acetylcholinesterase

The whole brain of the mouse was uniformly ground using a homogenizer, and then diluted to a concentration of 2 mg / ml in 0.1M, pH 7.4 phosphorylation solution, and used as an enzyme sample. Chlorogenic acid is sufficiently diluted in phosphorylated solution at concentrations of 5.5, 27.5, 55, 110, 220 and 440 ㎍ / ml, 1.5 ml of sample is added to 2.6 ml of phosphorylated solution, and 75 mM acetylthiocholine iodide 20 100 μl and 100 μl of Ellman reagent were added and reacted at room temperature for 30 minutes. 400 μl of enzyme sample was added thereto and the absorbance was measured at 412 nm. Inhibition capacity was calculated by the following equation using the measured absorbance values, and the experimental results are shown in FIG. 1.

% Inhibition = [1- (test group O.D.-test blank O.D.) / (Control O.D-control blank O.D.)] x 100

The acetylcholinesterase inhibitory activity of chlorogenic acid increased dose-dependently, showing 55, 62, and 78% acetylcholinesterase inhibitory activity at 110, 220, and 440 μg / ml, respectively.

Example 5 Free Radical Scavenging Activity of Chlorogenic Acid

DPPH (2,2-diphenyl-1-picrylhydrazyl, Sigma D9132) was prepared by dissolving 0.1 mM in 99.5% ethanol. Chlorogenic acid was diluted in ethanol at concentrations of 0.1, 1, 2.5, 5, 10 and 20 μg / ml. 1.8 ml of DPPH was added to 200 µl of the sample, mixed for 10 seconds, and reacted at room temperature for 30 minutes. The absorbance of the mixture was measured at 517 nm.

Using the measured absorbance values, the inhibitory capacity was calculated by the following equation, and the experimental results are shown in FIG. 2.

Inhibitory activity (%) = [(Control OD- Test Group OD) / Control OD] × 100

The free radical scavenging ability of chlorogenic acid increased in a dose dependent manner, and showed over 100% free radical scavenging ability.

Example 6 Working Memory Enhancement Effect of Chlorogenic Acid in Y-maze Experiment

Mice were divided into 4 groups, 10 of each group. The first group was the distilled water administration group, the second group, the third group and the fourth group were chlorogenic acid 3, 6 and 9 mg / kg administration group, respectively.

Chlorogenic acid was dissolved in distilled water and orally administered to mice at doses of 3, 6, and 9 mg / kg, and after 1 hour, the mice were placed in a Y-maze to measure the free entry of mice into branches A, B, and C. . At this time, each new branch is given 1 point (actual change), and the% equation for cross action is as follows.

% Cross action = (actual change) / (total number of entries-2) × 10

The test results are shown in FIG. 3.

Mice in the control group showed 60% crossover behavior on the test day, 3% / kg chlorogenic acid group showed 63%, increased 3% crossover behavior, and 6% / kg group showed 65% crossover behavior. The increase was 5%, while the 9 mg / kg group showed 68%, resulting in an 8% increase in crossover behavior. Therefore, it can be seen that by administering chlorogenic acid can significantly increase the working memory of the mouse. (p <0.05)

Example 7 Effect of Scopolamine-Induced Amnesia on the Y-maze of Chlorogenic Acid

Mice were divided into 5 groups, 10 of each group. The first group (control) was a distilled water administration group, the second group was a saline and scopolamine administration group, the third to fifth groups were chlorogenic acid 3, 6, 9 mg / kg and scopolamine administration group, respectively.

Chlorogenic acid was dissolved in distilled water and orally administered to mice at doses of 3, 6 and 9 mg / kg, and 0.5 mg / kg of scopolamine was injected subcutaneously after 30 minutes. After 1 hour, the mice were placed in the Y-maze to measure the free entry of the mice into branches A, B and C. At this time, each new branch is given one point, and the% equation for cross action is as follows.

% Cross action = (number of all three) / (total number of all-2) × 100

The test results are shown in FIG. 4.

The control group exhibited 64% and maintained spatial memory, but the scopolamine-treated control group showed 53% cross-behavior on the test day, leading to forgetfulness (p <0.01), and the chlorogenic acid 3mg / kg-treated group showed 56% cross-behavior. Increased by 3%, and the 6 mg / kg-administered group showed 63%, increasing mice's crossover behavior by 10% (p <0.01). The 9 mg / kg group showed 64%, which increased the crossover behavior of mice by 11% (p <0.01). Therefore, it can be seen that administration of chlorogenic acid significantly increases the working memory of reduced mice.

Example 8 Improvement of Scopolamine-Induced Forgetfulness in Underwater Maze of Chlorogenic Acid

Mice were divided into 5 groups, 10 of each group. The first group (control) was a distilled water administration group, the second group was a saline and scopolamine administration group, the third to fifth groups were chlorogenic acid 3, 6, 9 mg / kg and scopolamine administration group, respectively. On the outer wall of a circular grass 100 cm in diameter and 45 cm in height, four signs of stars, squares, triangles, and circles were attached to the wall, and a platform of 4.5 cm in diameter and 14.5 cm in height was placed. Water was filled up to 1 cm above the platform (water temperature 21 ± 1 ° C), and the food was used to make the platform invisible from the surface. We recorded the escape time to get to the platform after getting the mouse from the same position with the head facing the wall of the tank, and the cut-off time of the mouse was 120 seconds. Fifteen minutes later, the mice were regained and the escape time to reach the platform was recorded and compared with the control. The test results are shown in FIG. 5.

Mice in the control group showed an escape time of 44 seconds on the test day, scopolamine control group showed an escape time of 88 seconds, significantly inducing forgetfulness (p <0.001), and chlorogenic acid 3 mg / kg group showed 76 seconds The escape time was reduced by 12 seconds, and the 6 mg / kg group showed 62 seconds, which reduced the escape time of the mice by 26 seconds (p <0.05), while the 9 mg / kg group showed 46 seconds, reducing the escape time by 42 seconds. Second decrease (p <0.01). Therefore, it can be seen that the administration and memory of the mouse significantly increased due to the administration of chlorogenic acid.

In the exploratory experiments, mice in the control group showed a search time of 29 seconds in the target compartment, and scopolamine-treated control group showed a search time of 21 seconds, which significantly induced forgetfulness (p <0.05), and chlorogenic acid 3 mg / kg. The administration group showed 23 seconds, and the search time was increased by 2 seconds compared to the scopolamine administration control group. The 6 mg / kg group showed 26 seconds of search time and increased 5 seconds (p <0.05), and the 9 mg / kg group showed 28 seconds of search time. Memory for learning and learning was significantly increased (p <0.01).

There was no significant change in swimming speed at the 3 mg / kg and 9 mg / kg doses of the control, scopolamine control, and chlorogenic acid in the underwater lab experiment.

Figure 1 shows the ability to inhibit acetylcholinesterase by chlorogenic acid.

2 shows the results of free radical scavenging experiments with chlorogenic acid.

Figure 3 shows the working memory enhancement effect in the Y-maze experiment with chlorogenic acid.

4 shows the working memory (Y-maze experiment) enhancing effect of chlorogenic acid on scopolamine-induced forgetfulness.

5 shows the spatial memory (Morris water maze experiment) enhancing effect of chlorogenic acid on scopolamine-induced forgetfulness.

Claims (5)

A pharmaceutical composition for the prevention or treatment of learning or memory disorders comprising a pharmaceutically effective amount of chlorogenic acid or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. A pharmaceutical composition for the prevention or treatment of dementia comprising a pharmaceutically effective amount of chlorogenic acid or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The pharmaceutical composition according to claim 1 or 2, comprising 0.001 to 50% by weight of chlorogenic acid or a pharmaceutically acceptable salt thereof, based on the total weight of the composition. Learning disorders, comprising chlorogenic acid or a pharmaceutically acceptable salt thereof as an active ingredient, such that the chlorogenic acid or a pharmaceutically acceptable salt thereof is administered to the subject at a dose of 0.01 to 500 mg / kg per day, Drugs for the prevention or treatment of memory disorders or dementia. Food, health supplement or dietary supplement for improving learning ability or memory comprising chlorogenic acid or a pharmaceutically acceptable salt thereof as an active ingredient.

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