KR20150049324A - Composition for Improving Memory and Cognitive Function Comprising Arabinogalactan - Google Patents

Abstract

The present invention relates to a composition for improving a memory and cognitive function containing arabinogalactan, and more specifically, to a pharmaceutical composition for improving a memory or for preventing or treating cognitive dysfunction containing arabinogalactan as an active component, and the food composition for improving memory or for preventing or improving cognitive dysfunction. The composition containing arabinogalactan of the present invention has effects on improving memory and on improving and treating cognitive dysfunction such as dementia or the like, and thus can be effectively used as medicine, food, and others for improving memory or treating cognitive dysfunction.

Description

TECHNICAL FIELD The present invention relates to a composition for improving memory and cognitive function including arabinogalactan,

The present invention relates to a composition for improving memory and cognitive function comprising arabinogalactan, and more particularly, to a composition for improving or improving cognitive function or memory effect comprising arabinogalactan as an active ingredient, To a food composition for preventing or ameliorating memory or cognitive dysfunction.

Dementia is a complex clinical syndrome characterized by a marked impairment in human cognitive function, intellectual ability, emotional and behavioral changes. It causes cortical dysfunction such as memory, attention, language function, and spatio-temporal ability to lead daily life and social life It is in a state of great difficulty. The term "dementia" refers to a condition in which one or more of the other cognitive functions, including memory, is present.

Alzheimer's Disease or Parkinson's Disease is the cause of dementia, which can be caused by cerebral hemorrhage, metabolic diseases such as hepatic encephalopathy, Wilson's Disease, infectious diseases such as neurosyphilis, acquired immunodeficiency, Such as drug addiction, brain trauma, etc. In some forms, diseases that may cause structural and functional abnormalities in the central nervous system may cause dementia. Among them, dementia caused by Alzheimer's disease is the most common with 50 ~ 60%, followed by dementia caused by cerebrovascular disease.

It is estimated that the number of dementia patients among the elderly people aged 65 or older is about 460,000 in Korea, and that it will reach 700,000 in 2020 and 2 million in 2040 (Seoul National University Hospital). p. 132, 2008). Because dementia requires long-term treatment and the nature of its symptoms, the degree of life inconvenience is worse than other severe diseases such as cancer, and the physical, psychological, and economic burden of patient care and protection is also great. With the rapid rate of increase in the elderly population and the prevalence of dementia, it is clear that dementia is a task that requires social and national solutions.

Memory cognitive impairment is the first and most common symptom of Alzheimer's disease. In the early stages of Alzheimer's disease, patients suffer from recent memory disorders that do not remember the details of recent conversations or work, due to the impaired ability of hippocampal neurons to damage recent memory . But at this time, the remote long-term memory of events in the distant past is relatively well preserved. However, as the disease progresses, memory of the past gradually becomes disturbed as the cerebral cortex associated with the storage of long - term memory is damaged.

These symptoms of Alzheimer's disease are related not only to cytotoxicity by β-amyloid deposition but also to synaptic dysfunction of the cholinergic system (PM et al., Interactions between the amyloid and cholinergic mechanisms in Alzheimer's disease. Neurochem Int., 53: pp. 103-111, 2008). Cholinergic dysfunction is known to contribute to memory and cognitive dysfunction in Alzheimer's disease patients. Meynert's basal nucleus of Meynert Cholinergic neurons are associated with memory and cognitive function in association with temporal lobe, hippocampus and amygdala. In brain of Alzheimer's disease, 78% in temporal lobe, 60% in hippocampus, , It is known that neurons decrease by 67%. When brain cells are damaged by cytotoxicity, there is a barrier to the transmission of certain information, ie, the metabolism of neurotransmitters, which causes memory impairment. Many researchers have been steadily reporting the selective reduction of acetylcholine and its associated enzyme (choline acetyltransferase) in Alzheimer's disease. Furthermore, in the brains of patients with Alzheimer's disease, the nicotinic acetylcholine receptor, the muscarinic acetylcholine receptor, as well as the cholinergic reabsorption and acetylcholine secretion The function is also degraded.

Several cholinergic drugs have been developed to reverse these cholinergic system disorders, but the most effective and most widely used cholinesterase inhibitors are acetylcholinesterase inhibitors (AchEI). Currently marketed under FDA approval are donepezil, rivastigmine, and galanthamine, which inhibit the degradation of acetylcholine in synaptic clefts and increase the concentration, . These drugs show improvement in cognitive function and daily life in the early stages but they do not fundamentally prevent disease progression in that they return to the pre-administration state after 9 months to 1 year. The effect can be seen only if it is used relatively early, and it is not effective in the case of severe dementia. Common side effects include nausea, diarrhea, loss of appetite, dizziness, muscle cramps and sleep disturbances due to increased acetylcholine, and serious side effects include syncope. In addition, memantine (NMDA receptor blocker), ginkgo biloba extract (ginko biloba), huperzine A (reversible AChEI) extracted from Chinese medicine, The development of a variety of therapies related to the pathology of Alzheimer's disease, including the formation of protein, beta amyloid protein, deposition inhibition, antioxidant, antiinflammatory effect, etc., has been studied.

On the other hand, if the area in which the cerebral artery is clogged and the tissue is dying due to thrombosis following arteriosclerosis develops in a major region such as the hippocampus, it develops as dementia. The largest proportion of vascular dementia Or subcortical ischemic vascular dementia (SIVD) (35-67% of total vascular dementia) that occurs due to inadequate blood supply (Roman et al., Subcortical ischaemic vascular dementia. Lancet Neurol 2002; : 426-436). Damage to cortical ischemic vascular dementia (SIVD) occurs primarily in the white matter, and a variety of methods have been used to create animal models that resemble these diseases (Hainsworth & Markus, J Cerebral Blood Flow & Metabolism 2008; 28: 1877-1891). In vascular dementia, it has been shown that oligodendrocyte, which is a whitish material, is killed by apoptosis in the condition of vascular dementia, and studies are under way to suppress it. For example, edaravone, which is used as a stroke drug in Japan, and cilostazol, which is effective in stroke in an animal experiment, inhibit cell damage by inhibiting apoptosis in a rat BCCAO model (Ueno et al., Edaravone attenuates white matter lesions through endothelial protection in a rat chronic hypoperfusion model. Neurosci 2009; 18: 317-327).

At present, in the case of Alzheimer's disease treatment in Korea, foreign affiliated companies are highly dependent on imports so that they occupy a share of 98%. In addition, one company accounts for 80% of the market, which is virtually monopolistic (Korea Institute of Science and Technology Information, dementia treatment p.116, 2002). This shows that the growth of dementia treatment sector is urgent in terms of national competitiveness. Therefore, the development of effective anti-cancer drugs and memory-improving agents with fewer side effects can significantly improve the quality of life of Alzheimer's disease patients and improve and cure early-onset disease, as well as for amenorrhea and dementia patients suffering from memory cognitive impairment. It is thought that it can give.

On the other hand, arabinogalactan is industrially used as a thickener and stabilizer. Especially, it is a compound used as an emulsifying stabilizer in foods such as ramen, sausage, bread, etc. Recently, it has been reported that arabinogalactan There is no known effect on the nervous system although it is attracting attention. The present inventors have disclosed the use of arabinogalactan for the prevention or treatment of ischemic diseases through the prior patent (Korean Patent Application No. 10-2011-0121078).

Under these circumstances, the inventors of the present invention have found that arabinogalactan has an excellent effect for restoring memory impairment and treating cognitive dysfunction such as vascular dementia as a result of intensive efforts to reveal the memory effect and cognitive dysfunction of arabinogalactan The present invention has been completed.

It is an object of the present invention to provide a pharmaceutical composition containing arabinogalactan as an active ingredient for the prevention or treatment of memory improvement or cognitive dysfunction.

It is another object of the present invention to provide a food composition for preventing or ameliorating memory effect or cognitive dysfunction comprising arabinogalactan as an active ingredient.

In one aspect of the present invention, the present invention provides a pharmaceutical composition comprising arabinogalactan as an active ingredient for the prevention or treatment of memory improvement or cognitive dysfunction.

The term "arabinogalactan " in the present invention means a kind of water-soluble polysaccharide contained in the woody part, mud, sap, etc. of coniferous trees such as pine and Larix occidentalis NUTT or acacia, Galactose, and the composition ratio thereof is about 1: 6, and a galactose side chain of a plurality of β-1,6 bonds as a side chain and a galactose side chain of 1-2 ara It has a high structure in the form of coupled vinos. It exhibits high water solubility but is not soluble in ethanol, and has excellent salt resistance and pH stability. In natural state, it is mostly present in a small amount (less than 10%) of protein. Refined arabinogalactan exhibits a white or pale brown powdery state and is stored in a cool and dark place avoiding high temperature and humidity. In the case of an aqueous solution, it exhibits a lower viscosity (10% aqueous solution 25 ° C and 5 mPa · s or less) compared with other polysaccharide aqueous solutions.

It is industrially used as a thickener and a stabilizer. It is used as an emulsifying stabilizer in foods such as ramen, sausage, bread, and the like. Recently, the above-mentioned arabinogalactan has been attracting attention to the immune response and immunity-enhancing effects. However, there is no known effect on the nervous system, and for the purpose of the present invention, the arabinogalactan improves memory and cognitive dysfunction And the like, and the like.

The composition comprising arabinogalactan as an active ingredient of the present invention has an effect of improving memory and preventing, treating or improving cognitive dysfunction.

The term "memory" or "memory capacity" in the present invention means the ability to acquire new information, learned experiences, and knowledge obtained from the surrounding environment, to encode and store it in a specific region of the brain and to recall it. The composition of the present invention has the effect of improving the memory ability as described above. Such effects may include not only the effect of enhancing the normal person's memory but also all the effects of restoring the impaired memory or memory disorder.

The memory disorder refers to a state in which the memory is lowered compared to a normal person due to various causes such as trauma, attention deficit, aging, and disease, and includes forgetting, concentration disorder, loss of spatial perception, slowing of learning ability, Concept.

In an embodiment of the present invention, in order to confirm the effect of arabinogalactan on memory, passive avoidance experiments were conducted on mice induced memory impairment (forgetfulness) by scopolamine, and as a result, (Fig. 1). In addition, the corticosteroids inhibited cognitive memory damage by scopolamine (Fig. 1).

In the present invention, the term "cognitive function" is the ability to efficiently manipulate knowledge and information. For the purpose of the present invention, cognitive dysfunction refers to a condition in which the cognitive function is deteriorated, a disease caused thereby, As a concept that may be included, it may preferably mean a disorder resulting from damage of the cranial nerve cells. The cognitive dysfunction of the present invention may include forgetfulness, Alzheimer's disease, vascular dementia, dementia caused by head injury or Parkinson's disease, and preferably vascular dementia.

In an embodiment of the present invention, in order to confirm the effect of arabinogalactan on vascular dementia, the effect of the vascular dementia on the damage of the white matter using the chronic perfusion lowering model was examined. As a result, (Fig. 4), and further confirmed that arabinogalactan inhibited the activation of microglial cells, which are inflammatory cells of vascular dementia (Fig. 5), by inhibiting vinogalactan- It was confirmed that the arabinogalactan of the present invention has an effect of improving vascular dementia.

Preferably, the prevention, improvement and therapeutic effect of the memory improvement and cognitive dysfunction of the present invention may be achieved by increasing the expression of a brain-derived neurotrophic factor (BDNF). The BDNF plays a central role in nerve cell damage protection and nerve regeneration, and the effect of the present invention can be exhibited by the increased expression of BDNF. In one embodiment of the present invention, the expression of BDNF reduced by administration of scopolamine and the activity of its cAMP response element-binding protein (CREB) and phosphorylase (protein kinase B / Akt) (Figs. 2A and 2B).

Also preferably, the effect of preventing, ameliorating and treating the memory improvement and cognitive dysfunction of the present invention may be achieved by increasing the expression of choline acetyltransferase (ChAT). The ChAT is an enzyme that plays a central role in the synthesis of acetylcholine, and the effect of the present invention as described above can be exhibited by its increased expression. In one embodiment of the present invention, it was confirmed that the expression of ChAT reduced by administration of scopolamine was markedly increased by arabinogalactan (FIG. 3).

In the present invention, the term "prevention" means any action that inhibits or delays the occurrence, spread and recurrence of cognitive dysfunction by the administration of the pharmaceutical composition according to the present invention, and "treatment" Suspicion of cognitive dysfunction, and any behavior that alters or alleviates symptoms of an onset individual.

The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable" of the present invention means that it exhibits properties that are not toxic to the cells or humans exposed to the composition. Such carriers may be used without limitation as long as they are known in the art such as buffers, preservatives, wetting agents, solubilizers, isotonic agents, stabilizers, bases, excipients and lubricants.

In addition, the pharmaceutical 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 according to a conventional method have. Furthermore, it can be used in the form of an external preparation for skin in the form of ointments, lotions, spray agents, patches, creams, powders, suspensions, gels or gels. Examples of carriers, excipients and diluents that can be included in the composition of the present invention 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 and talc are also used. Liquid preparations for oral use may include various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used in suspension, liquid solutions, emulsions and syrups 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.

In the composition of the present invention, arabinogalactan according to the present invention may be contained in an amount of preferably 0.01 to 70% by weight, more preferably 0.1 to 50% by weight based on the total weight of the composition. However, the composition as described above is not necessarily limited thereto, but may vary depending on the condition of the patient, the type of disease, and the degree of progression.

Meanwhile, the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount " of the present invention means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, The type of disease, the severity, the activity of the drug, the sensitivity to the drug, the method of administration, the time of administration, the route of administration and the rate of release, the duration of the treatment, the factors including the drugs used concurrently or concurrently and other factors well known in the medical arts . The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

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. The dosage of the active ingredient in the pharmaceutical composition may vary depending on the route of administration, the severity of the disease, the age, sex, and weight of the patient, and may be administered once to several times per day. However, in order to obtain the desired effect, it is preferable to administer it at 0.01 mg / kg to 10 g / kg, preferably 1 mg / kg to 1 g / kg per day. The administration may be carried out once a day or divided into several doses. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

In another aspect, the present invention provides a method of preventing or treating memory improvement or cognitive dysfunction, comprising administering arabinogalactan to a subject in need thereof.

The term "individual" of the present invention means a monkey, a cow, a horse, a sheep, a pig, a chicken, a turkey, a quail, a cat, a dog, a mouse, and a mouse including a human who needs improvement of the memory, , Rabbit or guinea pig, and the diseases can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to the individual. The pharmaceutical composition of the present invention can be administered in parallel with existing therapeutic agents.

The term "administering" of the present invention means providing the patient with the desired substance in any suitable manner, and the administration route of the composition of the present invention may be administered through any conventional route so long as it can reach the target tissue have. But are not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrathecal, rectal. In addition, the pharmaceutical composition of the present invention may be administered by any device capable of moving the active substance to the target cell. The preferred modes of administration and formulations are intravenous, subcutaneous, intradermal, intramuscular, and drip injections. The injectable solution may be a non-aqueous solvent such as an aqueous solvent such as a physiological saline solution or a ring gel solution, a vegetable oil, a higher fatty acid ester (e.g., oleic acid), an alcohol (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.) (For example, ascorbic acid, sodium hydrogen sulfite, sodium pyrophosphate, BHA, tocopherol, EDTA and the like), an emulsifier, a buffer for pH control, a microbial growth inhibitor And a pharmaceutical carrier such as a preservative (e.g., mercury nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

In another aspect, the present invention provides a food composition comprising arabinogalactan as an active ingredient for preventing or ameliorating memory effect or cognitive dysfunction.

The arabinogalactan of the present invention is as described above, and the arabinogalactan of the present invention has the effect of improving memory and preventing or improving cognitive dysfunction, so that it can be included in the food composition as a food additive have.

 When the composition of the present invention is used as a food composition, the arabinogalactan can be used as it is, or can be used in combination with other food or food ingredients, and can be appropriately used according to ordinary methods. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment), and may further include a food-acceptable food-aid additive.

The food composition of the present invention may include all foods having a conventional meaning, and may be mixed with terms known in the art such as functional foods, health functional foods, and the like.

The term "functional food" in the present invention means a food prepared and processed by using a raw material or ingredient having functionality useful to the human body according to Law No. 6727 on health functional foods, and " And function of the nutrient for the purpose of obtaining a beneficial effect in health use such as controlling the nutrient or physiological action.

The term "health functional food" of the present invention refers to a food prepared by processing a specific ingredient as a raw material for the purpose of health assisting or by extracting, concentrating, refining, mixing, or the like a specific ingredient contained in a food raw material, Refers to a food which is designed and processed so that the body control function such as bio-defense, regulation of biorhythm, prevention and recovery of disease and the like can be sufficiently exhibited to the living body by the above components. Recovery, and so on.

There is no limitation on the kind of food in which the composition of the present invention can be used. In addition, the composition comprising arabinogalactan of the present invention as an active ingredient can be prepared by mixing other appropriate ingredients, which may be contained in food, with known additives according to the selection of a person skilled in the art. Examples of foods that can be added include dairy products, such as meat, sausage, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Vitamin complex, and the like, and can be prepared by adding to juice, tea, jelly, juice and the like prepared using the compound according to the present invention as a main component. It can also be used in the form of pills, powders, granules, infusions, tablets, capsules or drinks.

The health beverage composition of the present invention is not particularly limited to liquid ingredients other than the above-mentioned compounds as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages. 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, cyclodextrins; And sugar alcohols such as xylitol, sorbitol, and erythritol. As natural flavors other than those described above, natural flavors (such as tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and synthetic flavors (saccharin, aspartame, etc.) have. 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 fruit juice drinks 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 composition comprising arabinogalactan of the present invention exhibits an effect of preventing or ameliorating cognitive dysfunction such as memory improvement and dementia, and is useful for medicines and foods for the improvement of memory or cognitive dysfunction have.

Figure 1 shows the results of passive avoidance experiment showing the memory enhancing effect according to the concentration of arabinogalactan (AG) in mice in which memory damage was induced by scopolamine (SCO).
2A and 2B show Western blotting results (FIG. 2A) showing the degree of expression of BDNF (A), transcription factor CREB (B) and phosphorylated enzyme Akt (C) according to administration of arabinogalactan (Fig. 2B).
FIG. 3 shows RT-PCR results showing the degree of mRNA expression of choline acetyltransferase (ChAT) according to administration of arabinogalactan (AG) at different concentrations.
FIG. 4 shows the results of observing the degree of damage of aquatic plants by administration of arabinogalactan (AG) in brains (cc) and optically (opt) regions of vascular dementia-induced mice.
FIG. 5 shows the results of observing the degree of activation of microglial cells by administration of arabinogalactan (AG) in vascular dementia-induced mice.

Hereinafter, the constitution and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example  One: Arabinogalactan  Confirm memory effect

Arabinogalactan was obtained from Megazyme (Wicklow, Ireland) and dissolved in distilled water at a dose of 1 mg / kg, 2.5 mg / kg and 5 mg / kg, Respectively.

As experimental animals, C57BL / 6 male mice (7 weeks old, Hyochang Science, Daegu) weighing 20 ~ 25g were used for 1 week. The experimental group and the control group were divided into five groups. Arabinogalactan at doses of 1 mg / kg, 2.5 mg / kg and 5 mg / kg were orally administered during the adaptation period (7 days before the behavioral test) and during the behavioral experiment. The temperature of the experimental animals was maintained at 21 ~ 26 ℃ and the humidity was maintained at 40-60% in the animal breeding room of Keimyung University Medical School.

In order to confirm the effect of arabinogalactan on memory, we performed the following experiment using a passive avoidance test in mice that induced memory impairment by scopolamine.

Specifically, passive avoidance experiments were performed in a device in which bright compartments and dark compartments of the same structure (11 cm in width, 16 cm in height, and 10 cm in height) were connected by guillotine doors. The light compartment is equipped with a light bulb and the dark compartment is provided with a 2 mm thick stainless steel rods at 1 cm intervals on the floor to provide electrical stimulation.

The passive avoidance experiment was conducted for a total of 3 days. On the first day, I put my mouse on the bright compartment, adapted it for 20 seconds, turned on the light, and opened the compartment door. In general, the mouse enters a dark compartment without lighting, which automatically closes the compartment door. This training was repeated until the mouse entered the room without illumination in 20 seconds. On the second day, I put my mouse on the bright compartment, adapted it for 20 seconds, turned on the lights, and then opened the compartment door in the same way. When the mouse enters a dark compartment without lighting, the compartment door automatically closes and the electric current is applied through the net of the floor with a current of 0.5 mA for 5 seconds while the fire door is off.

On the third day of the experiment (after 24 hours), if the mouse is placed back into the bright compartment, the mouse will be reluctant to enter the dark compartment even when illuminated, and the time required for the mouse to enter the dark compartment (Retention trial). On the other hand, when the mouse did not enter the dark compartment, the cut-off time was set to 300 seconds.

As shown in FIG. 1, the step-through latency time was 237.86 ± 29.48 seconds in the normal group, and in the case of the SCO group in which the memory damage was induced by administering scopolamine, 27.53 ± 9.78 seconds, indicating a difference of 210.33 seconds. Thus, it was confirmed that memory damage was definitely induced.

On the other hand, in the experimental group in which arabinogalactan was administered at concentrations of 1 mg / kg, 2.5 mg / kg, and 5 mg / kg along with scopolamine treatment, 122.19 ± 46.61 seconds, 154.75 ± 43.19 seconds, and 217.3 ± 44.15 By showing the step-through latency time in seconds, the concentration-dependent administration of arabinogalactan was found to have a significant effect on the recovery of scopolamine-induced impaired memory (Fig. 1).

The results of this behavioral experiment showed that arabinogalactan is effective in restoring memory and enhancing cognitive function.

Example  2: Arabinogalactan  Confirming the increase effect of neurogenic expression

It was confirmed through the above Example 1 that arabinogalactan has an effect of improving cognitive memory, and in order to confirm the mechanism, arabinogalactan is a protein which plays a central role in nerve cell damage protection and nerve regeneration, derived neurotrophic factor (BDNF).

Western blotting was performed to measure the protein expression of the neurons and the activity of the cAMP response element-binding protein (CREB) and the phosphorylation enzyme (protein kinase B / Akt).

Specifically, the hippocampal portion was separated from the brain of the experimental animal after the behavioral experiment in Example 1, and the rats were divided into ice-cold RIPA buffer (150 mM NaCl, 0.5% Triton X-100, 50 mM Tris- mM NaF, 20 mM EGTA, 1 mM DTT, 1 mM Na ₃ VO ₄ , protease inhibitor cocktail). After lysis at 4 ° C for 20 minutes, 600 μl of supernatant was taken by centrifugation at 15,000 rpm for 15 minutes (Hanil, Korea). Protein content was quantitated with BCA (Bicinonic Acid; Pierce, USA) and 30 μg was electrophoresed on 12% SDS-PAGE. The developed gel was transferred to PVDF membrane (Pall Corporation, USA) and blocked with 5% fat-free dry milk-PBST (PBS, 0.1% Tween-20; AMRESCO, USA). Rabbit Anti-Goat IgG Conjugate (HRP-Rabbit Anti-Goat IgG Conjugate) was prepared by washing the cells with a primary antibody (AB1582, Chemicon, USA) overnight in 3% fat-free dry milk-PBS , 81-1620, ZYMED Laboratories, USA) were placed in 3% fat-free dry milk-PBS for 1 hour and then washed again with PBST solution for 10 minutes three times. After the reaction was carried out for 1 minute using a chemiluminescent solution (ECL; Amersham Pharmacia Biotech., IL, USA), it was photographed using a chemiluminescent immunoblotting imaging (UVP, CA, USA) Respectively.

As shown in FIGS. 2A and 2B, in the group administered with scopolamine alone, the BDNF (A), the transcription factor p-CREB (B) and the phosphorylated enzyme p-Akt (C) Expression was decreased, but the expression of arabinogalactan was significantly restored in the experimental group treated with 2.5 mg / kg and 5 mg / kg, respectively (Figs. 2a and 2b).

Example  3: Arabinogalactan  Increased expression of acetylcholine synthetase

To further investigate the mechanism of cognitive memory enhancement of arabinogalactan, mRNA expression of choline acetyltransferase (ChAT), an enzyme that plays a central role in the synthesis of the neurotransmitter acetylcholine, was confirmed Respectively. The expression of ChAT mRNA was tested by reverse transcription-polymerase chain reaction (RT-PCR).

Specifically, mRNA was extracted from the cerebral cortex of experimental animals with TRI Reagent (Molecular Research Center, OH, USA). The separated mRNA was reverse transcribed using M-MLV reverse transcriptase (Promega, WI, USA) at 42 ° C for 1 hour according to the manufacturer's protocol. The amplification of the cDNA was carried out by polymerase chain reaction (PCR) using a synthetic primer as described below. The DNAs amplified by PCR were electrophoresed on 1.5% agarose gel stained with EtBr (ethydiumbromide) and then examined with a gel documentation system (TCP-20.M, Vilber Lourmat, France) by UV irradiation . GAPDH (glyceraldehydes-3-phosphate dehydrogenase) was used as a control index to confirm the same experimental conditions.

The primer sequences for amplifying ChAT and GAPDH are shown in Table 1 below.

gene direction Primer sequence SEQ ID NO: ChAT Forward 5'-AGG GTG ATC TGT TCA CTC AG-3 ' One Reverse 5'-TCT TGT TGC CTG TCA TCA TA-3 ' 2 GAPDH Forward 5'-AGT GTA GCC CAG GAT GCC CTT-3 ' 3 Reverse 5'-GCC AAG GTC ATC CAT GAC AAC-3 ' 4

As shown in FIG. 3, the mRNA expression of choline acetyltransferase (ChAT) was decreased in the group administered with scopolamine alone compared with the normal group, but the arabinogalactan was administered at 2.5 mg / kg and 5 mg / kg, the expression was restored and markedly increased (FIG. 3).

Through these results, it was confirmed that arabinogalactan can increase the expression of neurotrophic factor and increase the expression of acetylcholine synthetase, thereby improving cognitive memory.

Example  4: Arabinogalactan  Due to vascular dementia White matter  Confirmation of damage inhibition effect

In the following experiments, the following samples and experimental animals were used.

Arabinogalactan was received from Megazyme (Wicklow, Ireland) and mixed with feed at a dose of 100 mg / kg / day, allowing rats to freely eat 5 days before ligation of the entire carotid artery.

Sprague Dawley male rats weighing 260-270 g (8 weeks of age, Sam Taco Bio Korea, Osan, Korea) were used as experimental animals for one week. (Sham), control group, and experimental group. Arabinogalactan was mixed with feed at a concentration of 100 mg / kg / day during the adaptation period (5 days before the behavior test) and 30 days of experiment to allow rats to eat freely. The experimental animals were maintained at 21 ~ 26 ℃ and 40 ~ 60% humidity in the animal breeding room of Daegu Catholic University Medical School.

In order to confirm the effect of arabinogalactan on vascular dementia, the inhibition effect on white matter damage was confirmed through a chronic hypoperfusion model commonly used for the application of bilateral common carotid artery occlusion (BCCAO) Respectively.

In the chronic perfusion-depleted model, when both carotid arteries of a rat are bound, the blood supplied through the vertebral artery is supplied to the brain through the circle of Willis. However, the amount of blood to be supplied is low and the white matter is selectively killed. There are corpus callosum, internal capsule, and optic tract in this region (Farkas et al., Permanent, bilateral common carotid artery occlusion in the rat: a model for chronic cerebral hypoperfusion-related neurodegenerative diseases. Brain Res Rev 2007; 54: 162-180). In addition to neurons composed of cell bodies and axons, the brain also contains astrocyte that nourishes nerve cells and removes excess neurotransmitters, and axons that make up nerve cells. (Oligodendrocyte), which produces the myelin necessary to facilitate the transmission of electrical signals through the microglial cells (oligodendrocyte) and microglial cells that act to remove dead cells when nerve cells die or become infected microglia) (Nature 2009 Feb 5; 457 (7230): 675-7), and the extracellular matrix consists mainly of the polysaccharide hyaluronan (Allen & Barres, Neuroscience: (Rauch, Extracellular matrix components associated with remodeling processes in brain. Cell Mol Life Sci 2004; 61: 2031-2045). Among them, the white matter involved in vascular dementia was mainly composed of axons of nerve cells. Therefore, the tissues were observed mainly in white color and the luxol fast blue staining was performed to confirm these damages.

Specifically, tissue slides were obtained at -2.64 to -3.12 mm from the bregma point at which white matter was present, stained with a Luxolfast blue solution and a lithium carbonate solution, (Piedio et al., A modified technique for high-resolution staining of myelin. J Neurosci Methods 2006; 153: 135-146). The converted scores were as shown in Table 2 below, and were observed mainly in the brains cc and opt.

Grade Degree of damage 0 normal One Disarrangement of nerve fibers 2 In the formation of marked vacuoles, 3 The disappearance of myelinated fibers

As shown in FIG. 4, it was confirmed that the score of damage was very high in the control group in which vascular dementia was induced compared to the normal sham group in which vascular dementia was not induced, whereas arabinogalactan AG) was significantly reduced in the group treated with 100 mg / kg than in the control group. The damage scores were 1.45 ± 0.43 (AG) and 1.89 ± 0.34 (control) in visual cortex (cc) with 1.06 ± 0.22 (AG) and 1.79 ± 0.11 (control) It was confirmed that administration of arabinogalactan inhibits the damage of white matter caused by vascular dementia, and arabinogalactan is effective in improving vascular dementia.

Example  5: Arabinogalactan  Confirmation of microglial cell activation inhibitory effect

In Example 4, it was confirmed that arabinogalactan had the effect of restoring white matter by vascular dementia, and further, the effect of inhibiting the activation of microglial cells, inflammatory cells of vascular dementia, was confirmed.

Activated oxygen and microglial cells activated by dead cells secrete not only dead cells but also cytokines such as active oxygen species, glutamate and IL-1, which are neurotransmitters, Again, oligodendrocyte is killed by cell suicide (Butts et al., Maturation-dependent sensitivity of oligodendrocyte lineage cells to apoptosis: implications for normal development and disease. Cell Death and Differentiation 2008; 15: 1178-1186 ; Dheen et al., Microglial activation and its implications in the brain diseases. Curr Med Chem 2007; 14: 1189-1197). Micro-glia cells are in the form of protuberances in normal state, but when activated, they become proliferated in the form of amoebas without protuberances (Dheen et al., 2007; Garden & Muller, 2006). Iba1 (ionized calcium binding adapter molecule 1) was used as an antibody for selectively staining microglial cells.

As in Example 4, the brain tissue slices of the paraffin-treated paraffin obtained at a thickness of 5 μm were treated with 0.03% H ₂ O ₂ and citric acid, followed by addition of 1% BSA (bovine serum albumin) and 5% normal serum Lt; / RTI > After that, the microglial cells were subjected to primary antibody attachment using Iba1 (diluted 1: 200, Wako, Osaka, Japan). After washing, secondary antibody (goat anti-rabbit IgG, pAb (HRP conjugate) diluted 1: 500, ELS, Lausen, Switzerland) was applied to Iba1 for 1 hour at room temperature. Then, they were reacted with avidin-biotin peroxidase complex solution using Elite Vectastain ABC kit (Vector Laboratories). Then, the solution was reacted with a solution containing 0.05% diaminobenzidine (Vector Laboratories) so as to develop a brown color. After dehydration, the tissue was stored. In the negative control group, IgG (immunoglobulin G) was attached instead of the primary antibody, and the whole process was performed in the same manner. After that, the analysis was conducted focusing on the (opt) area where the degree of damage was remarkably observed.

As a result, microglial cells were activated in the control group which induced vascular dementia compared to the normal group (sham), and the size of the microglial cells became larger, the number of the outstretched form was increased, and the intensity of the staining was also strongly developed. On the other hand, in the group treated with arabinogalactan (AG), activation of microglial cells was inhibited compared to the control group, and no protruding form was observed. As a result of quantitative analysis reflecting the intensity and area of staining, it was confirmed that the activity of microglial cells was reduced by about 45% in the group administered with arabinogalactan, compared with the control group, and arabinogalactan It means that vascular dementia can be improved.

Hereinafter, an example of the preparation using arabinogalactan will be illustrated, but the present invention is not intended to be limited thereto but will be specifically described.

Formulation example  One: Sanje  Produce

Arabinogalactan 300 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

Arabinogalactan 300 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

Arabinogalactan 150 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

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 injection

Arabinogalactan 150 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ .2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

Formulation example  5: Liquid  Produce

Arabinogalactan 150 mg

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added and dissolved in purified water according to the usual liquid preparation method, 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 added with purified water to adjust the total volume to 100 ml, And sterilized to prepare a liquid preparation.

Formulation example  6: Manufacture of health food

Arabinogalactan 1000 mg

Vitamin mixture quantity

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

0.2 [mu] g vitamin B12

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

50 ㎍ of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 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

Arabinogalactan 150 mg

Vitamin C 15 g

Vitamin E (powder) 100 g

19.75 g of ferrous lactate

3.5 g of zinc oxide

Nicotinic acid amide 3.5 g

Vitamin A 0.2 g

Vitamin B1 0.25 g

Vitamin B2 0.3 g

Water quantification

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the health beverage composition of the invention.

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.

<110> CATHOLIC UNIVERSITY OF DAEGU INDUSTRY ACADEMIC <120> Composition for Improving Memory and Cognitive Function          Comprising Arabinogalactan <130> PA130887 / KR <160> 4 <170> Kopatentin 2.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> ChAT Forward Primer <400> 1 agggtgatct gttcactcag 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> ChAT Reverse Primer <400> 2 tcttgttgcc tgtcatcata 20 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH Forward Primer <400> 3 agtgtagccc aggatgccct t 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH Reverse Primer <400> 4 gccaaggtca tccatgacaa c 21

Claims (6)

A pharmaceutical composition for preventing or treating memory improvement or cognitive dysfunction, comprising arabinogalactan as an active ingredient.
The composition according to claim 1, wherein the cognitive dysfunction is any one selected from the group consisting of forgetfulness, Alzheimer's disease, vascular dementia, dementia caused by head injury, or Parkinson's disease.
The composition of claim 1, wherein the prevention or treatment of memory improvement or cognitive dysfunction is achieved by increased expression of a brain-derived neurotrophic factor (BDNF) or choline acetyltransferase (ChAT) .
The composition of claim 1, wherein the arabinogalactan is present in an amount of 0.1 to 50% by weight relative to the total weight of the composition.
A food composition for preventing or ameliorating memory or cognitive dysfunction, comprising arabinogalactan as an active ingredient.
The composition according to claim 5, wherein the cognitive dysfunction is any one selected from the group consisting of forgetfulness, Alzheimer's disease, vascular dementia, dementia caused by head injury, or Parkinson's disease.

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