KR100456418B1 - Food materials of glycine binding site inhibition activity, food materials for preventing dementia and foods using the same - Google Patents

Abstract

The present invention is a food material for preventing dementia-related diseases prepared by combining alone or mixed with hot water extracts or concentrates of aged, acid, jinjin, fish vinegar, ogapi, huanggi, and engineering workers to enhance glycine binding inhibitory activity in patients with dementia. It can be used in the manufacture of various foods, and in particular, by adding them during the manufacturing process of the confectionery, dieting these confections can bring about prevention and improvement of various dementia-related diseases, as well as reduce the incidence of dementia patients in the aging age. The present invention relates to a method for producing a functional food for preventing dementia-related diseases.

Description

Food materials that enhance glycine binding inhibitory activity, functional food materials for preventing and improving dementia using the same and foods using the same {Food materials of glycine binding site inhibition activity, food materials for preventing dementia and foods using the same}

The present invention is prepared by using a variety of herbal extracts exhibiting glycine binding site inhibition activity, and then adding these compositions during the manufacturing process of various foods, including the traditional fruit family to generate high frequency in an aging society The present invention relates to a functional food having an effect of preventing and ameliorating dementia-related diseases.

Dementia is a brain disorder that indicates a general cognitive impairment in a clear state of consciousness, usually caused by chronic or progressive brain disorders, and impaired in many senior cerebral functions such as memory, thinking ability, understanding, calculation, learning, and language judgment. to be. Dementia is the most common disease in which human deaths rank third in the world after coronary artery disease and cancer. Around 25 million people worldwide suffer from this disease.In 1960, out of a total of 25 million people, the elderly population was 720,000, 2.9%, but in 1990, about 2.12 million (5.0%), 1995 It is expected to become a full-fledged aging society as it has increased to about 2.5 million people (5.6%) and from 2000 to 7-8% of the total population. Therefore, the number of patients with dementia increases rapidly with age, with 6-8% of patients over 65 years old and 30% of patients over 85 years old.

Dementia-induced diseases include senile dementia, cerebrovascular dementia, Pick's disease, Creutzfeldt-jakob's disease, and Parkinson's disease. Dementia, dementia, hypothyroidism, normal pressure hydrocephalus, diabetes, drug addiction, anemia and vitamin deficiency. Geriatric dementia accounts for 50 to 60% of adult dementia in the West, with severe diffuse brain atrophy and loss of brain cells, resulting in loss of memory and intelligence, impaired abstract thinking and high cortical function. It is a symptom showing delusion and behavioral disorders, and it is a representative disease showing disorder of higher mental function and desolation of personality. In addition, cerebrovascular dementia caused by extensive brain lesions after stroke or by cerebral atherosclerosis is reported to be relatively higher in the East than in Europe.

In Oriental Medicine, it was the first to refer to the symptoms of dementia in terms of dementia. Since then, it has been referred to as Seokbinbinok, dialectic, and so on in terms of symptoms and treatment. The records were relatively detailed. Before that, dementia was identified by including other symptoms similar to dementia such as mania, forgetfulness, depression, phlegm, and erosion. I think. In terms of treatment, with the exception of some of the aforementioned treatable dementia, senile dementia, pick disease, and cerebrovascular dementia have not been identified. Therefore, dementia may be more effective than preventive treatment.

Many drugs have been studied for the treatment of dementia, but no drug has yet been proven effective. However, in addition to the hypothesis that cognitive dysfunction is mainly caused by damage to choline neurons at the base of the cerebral base, choline drugs with various mechanisms have been developed, and some have proven somewhat effective. There is a bar. Recently, donepezil, which has high receptor specificity, long half-life, and simple administration method, has been developed and used in Europe and the United States. In addition, possible methods for the next generation of therapies include a combination therapy of drugs acting on various neurotransmitters, nerve growth factors, and genetic engineering treatments. In other words, in addition to choline neurons, various neurotransmitter abnormalities and cerebral cortical atrophy have been reported, so the pathology of dementia is not limited to one neurotransmitter or part of the brain, but is specific to a wide range of brain regions and nerves. Combination therapy of choline / serotonin, choline / somatostatin-containing drugs with delivery agents is expected. Considering the low regenerative capacity of neurons, it has recently been recognized that clinical methods are needed to delay the degenerative process. One of these roles is the nerve growth factor, which has been shown to protect the degeneration of choline neurons that occur after surgery in animal experiments and to the growth, regeneration and maintenance of choline neurons. It is an essential protein. There is still a problem in the method to be administered directly to the brain, but if this problem is solved, it is expected to attract attention as a new treatment method. Recently, it has been confirmed that a gene causing dementia exists on chromosome 21, and studies to identify genes of beta-amyloid protein, one of the important neurological lesions, have been actively conducted to determine the possibility of gene therapy for the disease. I have suggested.

Therefore, the most reasonable new material for preventing and improving dementia should be a drug that delays the destruction and aging of brain cells, restores cognitive function, protects brain cells and promotes the regeneration of brain cells. There is no current. Therefore, it is necessary to appropriately select the drug according to the symptoms and causes of the patient, but the reality is that it is left to the doctor's skill. Currently used drugs include antipsychotics (Antipsychotics), antidepressants (Annticepressant), antianxiety (Antianxiety), anticonvulsant (Anticonvulsant), brain stimulants (Nootropics), enzyme blockers involved in the metabolism of neurotransmitters (Enzyme blocker), Calcium blockers (Ca-blocker), etc. are used, and the present comparatively definite therapeutic effects are acetylcholinesterase inhibitors among choline nervous system agents. Memory impairment is an early symptom of Alzheimer's disease and is a representative symptom. This memory impairment is known to be due to the decrease of acetylcholine, and after data collected through brain image study of dementia patients, severe degeneration of acetylcholine nervous system These are drugs that have been developed because of the therapeutic process that is focused on.

As described above, one of the rational methods for preventing and improving senile dementia-related diseases in the development of a medicinal treatment for dementia is preventing and improving food materials, concentration and memory disorders that prevent and improve motor, sensory, and mental disability. To develop a composition using a food material, and a food material that is active in preventing the degeneration of the central nervous system, and then added to the food to take effect by ingesting a certain amount, therefore the present invention is the above element, that is, exercise, The present invention relates to a method for producing traditional Korean foods, in which a new food material with active sensation, persistence, concentration, memory, and prevention of central nervous system is developed, the composition is prepared in combination with the material, and the overall activity of the above factors is exerted. .

An object of the present invention is to provide a food material using the hot water extract of the herbal medicine to enhance the glycine binding inhibitory activity.

In addition, an object of the present invention is to provide a functional article for preventing and improving dementia by using the food material.

The present invention is to inhibit the muscarinic M1 receptor binding so as to exhibit a comprehensive function in functional food materials and dementia-related diseases to enhance glycine binding inhibitory activity using hot water extracts of aging, mountain, jinjin, eoseongcho, ogapi, astragalus The present invention relates to a food material for enhancing activity, a food material for enhancing acetylcholinesterase inhibitory activity, and a food material for improving dementia, and a functional food material for improving dementia.

In another aspect, the present invention relates to a functional food for preventing and improving dementia, which is effective in preventing and improving dementia by adding the food material in the manufacturing process of the Korean medicine using the herbal food material. It can be added to the food preparation by the health food material or functional food material by known methods using hygiene law or pharmacologically acceptable prosthetic materials.

The herbal food material used in the present invention may be added in various formulations (extract, juice, hot water extract, powder, granules, grinding) according to the purpose.

The Chinese family referred to in the present invention includes all types of traditional family currently available on the market.

Functional food material for the prevention and improvement of dementia of the present invention and a method of manufacturing a confectionery using the same Example 1 below: Manufacturing method of functional food material for prevention and improvement of dementia, Example 2: Functional Chinese medicine method for preventing and improving dementia By way of example, the present examples should not be regarded as limiting the present invention.

Example 1. Method of manufacturing food material for preventing and improving dementia

<The first process>

Sewage to enhance the inhibitory activity of rhubarb, yellowish white, gaza, and acetylcholinesterase that enhance muscarinic M1 receptor binding activity. · 100g each of the above-mentioned materials of gold and silver, bark skin, killing, non-leafing leaves, celery, gardenia, and anti-oxidant active materials such as far wind, sand, soft potato, three hundred seconds, and bokbun, washed well with water, and then put 1500 ml of water in 95 ~ 100 ℃. Extract for more than 5 hours.

<The second process>

The hot water extract was filtered through a filter paper (whatman No. 3) to obtain 800 ml of pure herbal hot water extract, each of which was freed from debris.

<The third process>

Each filtered herbal extract is concentrated completely at 60 ° C. using a vacuum concentrator to obtain a powder.

<The fourth process>

The powder is diluted to a concentration of 0.005, 0.05, 0.5, 1.0 mg / ml with distilled water, and the activity of each herbal material mentioned in the first step is measured to determine whether dementia-related activity is detected. The results of various physiological activities for each herbal material show excellent dementia related activity as shown in Tables 1, 2 and 3.

<The fifth process>

Based on the results of the dementia-related physiological activity in the fourth step, aging, potions, injin, eoseongcho, ogapi, astragalus, 40g each of construction workers, 80g of rhubarb, 50g each of Gaza, sewage, gold silver flower, bark skin, kill, non-leaf leaves 10 liters of water was added to 910 g of the Chinese herbal medicine material, cheongung, gardenia gardenia, each 40g, and 50g each of the original wind, sagan, lotus root, three hundred seconds, and bokbunja, followed by hot water extraction at 95 ~ 100 ℃ for 5 hours. The concentrated solution 1200ml (40Brix) is obtained through the 3rd process.

<The sixth process>

The concentrated solution prepared in the fifth step is partially lyophilized and used for investigation of dementia-related activities, and the rest is used for the manufacture of functional Chinese medicine for preventing and improving dementia. The lyophilized product prepared as described above was found to have high related activity as shown in Table 4 when the dementia related activity was measured by diluting to 0.005, 0.05, 0.5, 1.0 mg / ml with distilled water.

Table 1. Glycine binding site inhibition activity for each herbal extract

Medicine Concentration (mg / ml) % Inhibition Heraldic 0.0050.050.51.0 5586100100 Acid pills 0.0050.050.51.0 5795100100 Jin 0.0050.050.51.0 4173100100 Air castle candles 0.0050.050.51.0 4071100100 Cuckold with feet 0.0050.050.51.0 4271100100 Yellow 0.0050.050.51.0 317894100 Public sign 0.0050.050.51.0 3473100100

Table 2. Muscarin M1 receptor binding inhibition activity for each herbal extract

Medicine Concentration (mg / ml) % Inhibition Yellow lotus 0.0050.050.51.0 4193100100 Sulfur bag 0.0050.050.51.0 01391100 Go 0.0050.050.51.0 03467100

Table 3. Acetylcholinesterase inhibition actibity for each herbal extract

Medicine Concentration (mg / ml) % Inhibition Sewage 0.0050.050.51.0 1352100100 Gold silver coins 0.0050.050.51.0 8528897 Throat 0.0050.050.51.0 01585100 Kill 0.0050.050.51.0 0147698 Non-cotyledon 0.0050.050.51.0 04279100 Celestial palace 0.0050.050.51.0 07087100 Gardenia 0.0050.050.51.0 034100100

Table 4. Dementia related activities of the herbal extract composition of the sixth step of Example 1

activation Concentration (mg / ml) % Inhibition Glycine binding inhibitory activity 0.0050.050.51.0 56100100100 Muscarinic M1 receptor binding inhibitory activity 0.0050.050.51.0 4887100100 Acetylcholinesterase Inhibitory Activity 0.0050.050.51.0 6491100100

The results of Table 1 and Table 2 study the effect of each drug on the glycine binding site inhibition (Mlycarine M1 receptor binding inhibition) activity and the relationship between the receptor-ligand In order to react with the receptor using a ligand attached to the radioisotope, the resultant is filtered through a glass fiber filter to remove the unbound extra ligand and then remains on the washed filter disc. The amount of the isotope was measured to quantify the binding reaction of the ligand to the receptor and to determine the effect of the drug.

Receptor sources for the present invention include human recombinant muscarin receptor subtypes 1 and 2 expressed in CHO cells to exclude interactions with various subtypes of each drug. M ₁ , M ₂ ) and glycine binding site (glycine binding site) receptor was used as a fraction of the receptor isolated from the rat brain. Cell fractions containing large amounts of these receptors were stored frozen and diluted to the optimal concentration.

Acetylcholinesterase inhibitory activity of Table 3 is obtained by reacting the product obtained by reacting thiocholine released from acetylcholine with DTNB (5,5'-dithio bis-2-nitrobenzoate). The activity was calculated by measuring.

In other words, the muscarine M1 receptor binding inhibition activity was suspended in -70 ℃ receptor cell fractions stored in the buffer for each experiment and the protein content of the Bio-Rad protein synthesis kit (Bio-Rad DC Protein Assay Kit) and then adjusted to the optimal concentration for each receptor. At this time, the protein concentration represented by the content of the receptor is determined through the basic experiment. Samples of all tests were replicated and each buffer was used as the buffer required for the experiment. The final volume of the reaction was 0.25 ml, which contained 50 μl of hot-ligand and 10 μl of test drug. The reaction was started for 30-60 minutes at 27 ° C. by adding 100 μl of receptor suspension. In the first step drug search, the affinity of the drug for receptors was searched for two concentrations (1 μM and 10 μM). After incubation, the reaction was terminated by filtering with cold Tris buffer using an Inotech cell harvester system using a Wallac glass fiber filtermat GF / C. Isotopes bound to the receptors were isolated, washed, and the amount of isotopes remaining in the filtermat was measured with a MicroBeta-Counter. Recent developments in cholinergic drugs (cognitive functioning agents) In addition, research to control other degenerative brain diseases such as dementia and cerebral ischemia by preventing brain cell damage caused by various excitatory amino acids such as glutamate and NMDA (N-Methyl-D-Aspartate) has attracted new attention, especially excitatory neurotransmitters. There is an active research to develop selective antagonists of glutamate receptors. Only one day excitatory neurotransmitters (excitatory neurotransmitter) as it is known to have also closely associated with corruption of various cognitive functions such as learning and memory associated with dementia. In addition, in order for glutamate to cause damage to brain cells, binding to glutamate receptors must be preceded first. To date, more than four glutamate receptors have been identified, of which metabotropic glutamate receptors are involved in the transmission of various signals through the intracellular second messenger cascade, and the other types of receptors are called ionotropic receptors because they represent the function of ligand-gated ion channels. In addition, AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate) receptor and kainate receptor are well known. NMDA receptors trigger Ca ++-dependent signaling mechanisms in neurons. In other words, NMDA receptors act as the most important Ca ++ channels in neurons and they promote the influx of Ca ++ into neurons by various NMDA agonists such as NMDA, which induces apoptosis of neurons and this apoptotic process is an NMDA antagonist. In order to measure the activity of these NMDA antagonists, selected samples were selected as ligands for the NMDA receptor fraction isolated from the rat brain and the glycine binding site of the receptor. The affinity to the NMDA receptor was searched for as an indicator of the effect of inhibiting binding to known [3H] -MDL 105,519. Meanwhile, glycine binding inhibitory activity was measured by the following method. (1) Reagent and instrument Liquid scintillation counter MicroBeta 1450 Plus (Wallac, Finland) was used as the Inotech harvester (96-well) and shaking incubator (Rosi 1000, Thermolyne) was used. Ligand reagent [3H] MDL 105,519 used for receptor affinity testing was purchased from Amersham Pharmacia Biotech, and NMDA antagonist drug used as a reference drug was 5,7-DCKA (5,7-Dichlorokynurenic acid, IC50 value for Glycine binding site is estimated as 1.00 μM) was purchased from RBI. (2) Separation of receptors The NMDA receptors used in the experiments were isolated by synaptic membranes from the entire brain of the male rat (Spague-Dawley). In other words, the whole brain of the rat (Spague-Dawley) was extracted, chopped, and added with a 10-fold volume of cold sucrose solution (0.32 mM), homogenized using a Teflon-glass homogenizer and immediately 1,000 g (10 minutes, 4 ° C, The supernatant was obtained by centrifugation with a Beckman J2-21 M / E centrifuge. The supernatant was centrifuged again at 20,000 g (20 min, 4 ° C.) to obtain a precipitate. 20 times of cold distilled water was added to the obtained precipitate, homogenized with Brinkman Polytron Homogenizer for 30 seconds, stirred at 4 ° C for 30 minutes, and then centrifuged at 8,000 g (20 minutes, 4 ° C). The supernatant was taken off. The precipitate obtained by centrifuging the supernatant again with 39,800 g (25 min, 4 ° C., Beckman L8-M ultracentrifuge) was cryopreserved at −70 ° C. The cryopreserved precipitate was dissolved at room temperature for 10 minutes and then homogenized in 50 mM Tris-acetate buffer (pH 7.1) containing 20 times the volume of 0.04% Triton X-100, which was stirred at 37 ° C. for 20 minutes and then 39,800 g (20 Minutes, 4 ℃) again centrifuged to obtain a precipitate. The obtained precipitate was washed three more times (homogenized and centrifuged) with a 20-fold volume of 50 mM Tris-Acetate buffer (pH 7.1) and then suspended in Tris-Acetate buffer to increase the protein concentration according to Bradford's method. After the measurement, the protein concentration was divided into 1 mg / ml, and stored at -70 ° C, which was dissolved in redemption and used as a receptor. (3) Receptor affinity test The receptor fraction, which was frozen at -70 ° C, was stored in 50 mM tris-acetate. The protein content was adjusted to 5 μg / well concentration by suspension with buffer (pH 7.1). 50 mM tris-acetate (pH 7.1) was used as the assay buffer. The final volume of the reaction solution was 0.25 ml and 50 µl of hot-ligand 4 nM [3H] MDL 105,519 (140,000 DPM) and the herbal extracts obtained in the fourth step of Example 1 were 0.005, 0.05, 0.5, 1.0 mg / ml. To be included. In addition, 50 μl of 5 mM glycine was added to correct nonspecific binding. Initiation of the reaction was carried out in a shaking incubator for 30 minutes at 25 ° C. after adding 100 μl of receptor suspension. After incubation, 0.2 ml of cold 50 mM Tris-HCl in 0.9% saline, (pH 7.4) was added to terminate the reaction. Immediately filtered with an Inotech cell harvester system using Wallac glass fiber filtermat GF / C (Wallac, PO Boc 10, FIN-20101 Tutku, Finland) and washed 9 times with cold buffer. After drying the filtermat in the microwave oven, radioactivity was measured by a liquid scintillation counter to calculate the binding ratio of ligand to the receptor. Each herbal sample was dissolved in a small amount of dimethylsulfoxide (DMSO), diluted with buffer, and the concentration of DMSO in the reaction solution was less than 0.1%. All samples were measured by duplicate and the average value was calculated. As a standard control drug, 5,7-DCKA (5,7-Dichlorokynurenic acid) was purchased from RBI. 5,7-DCKA was observed to inhibit the binding of ligand to the receptor by 50% at 1.0 μM concentration.

In the acetylcholinesterase inhibitory activity experiment, enzyme activity was produced by reacting thiocholine, which is released by hydrolysis of acetylcholine, which is a substrate, according to Elman's method, with DTNB. Enzymatic activity was measured by UV detection of dithio compounds and thio anions of nitrobenzoic acid. At this time, all reactions were fixed at 25 ° C. in a 1 ml cuvette to measure initial rates for 30 seconds and 60 seconds, and enzyme inhibition activity was calculated in terms of the initial rate ratio of the inhibition reaction to the reference reaction.

Example 2. A functional Korean confectionery for preventing and improving dementia

<The first process>

Selected glutinous rice (65%) is naturally fermented in water for about 2 weeks, washed well and ground to fine powder.

<The second process>

Jeongjong (1%), ginger juice (0.9%), refined salt (0.1%) is added to the glutinous rice kneaded well and steamed at 100 ℃.

<The third process>

Put steamed beans in the second process and add soy flour (19%) while striking to reduce the dough.

<The fourth process>

The raw material kneaded in the third step is put into a molding machine and then molded, and dried to 27% moisture at 27 ° C.

<The fifth process>

The dried raw material is fried in soybean oil (2%).

<The sixth process>

The confectionery fried in the fifth step is immersed in about 30 seconds to 1 minute in the syrup added to the herbal composition concentrated composition prepared in the fifth step of Example 1 at a concentration of 3 to 15%.

<The seventh process>

In the sixth step, coat the Korean confectionery coated with the herbal composition concentrated with white rice, and then dry it and package it.

The present invention is a food material for the prevention of dementia-related diseases by preparing a single or mixture of hot water extracts or concentrates of deciduous, acid, jinjin, effervescent vinegar, ogapi, huanggi, engineering workers to enhance glycine binding inhibitory activity in patients with dementia. It can be used in the manufacture of various foods, and especially by adding these foods in the manufacturing process of the Chinese medicine to prevent and improve various dementia-related diseases by dieting these foods or Korean fruits. Can be reduced.

Claims (9)

Extract water at 95-100 ° C. for at least 5 hours by adding water to herbal medicines selected from the group consisting of fermented soybean, medicinal herb, injin, eoseongcho, ogapi and corporation; An extract or a mixture of extracts prepared by concentrating the extract under reduced pressure; Food material to enhance the glycine binding inhibitory activity, characterized in that. The method of claim 1, wherein the selected herbal medicine, Food material to enhance the glycine binding inhibitory activity characterized in that it further comprises a sulfur group. delete delete (1) a food material that enhances the glycine binding inhibitory activity of paragraph 1 or 2, (2) adding water to the herbal medicine selected from the group consisting of yellow lotus, yellow white, and Gaza and extracting at 95-100 ° C. for at least 5 hours; An extract or a mixture of extracts prepared by concentrating the extract under reduced pressure; Food materials to enhance the muscarinic M1 receptor binding inhibitory activity, characterized in that (3) extract water at 95 ~ 100 ° C. for at least 5 hours by adding water to a herbal medicine selected from the group consisting of sewage, sterling silver, bark skin, killing, non-leafing, celestial and gardenia; An extract or a mixture of extracts prepared by concentrating the extract under reduced pressure; Food materials to enhance the acetylcholinesterase inhibitory activity, characterized in that (4) adding water to the herbal medicine selected from the group consisting of far-wind and bokbunja and extracting at 95-100 ° C. for at least 5 hours; An extract or a mixture of extracts prepared by concentrating the extract under reduced pressure; Food material to enhance the antioxidant activity, characterized in that Functional food material for dementia prevention and improvement, characterized in that comprises a. According to claim 5, Herbal medicine selected for the production of food ingredients that promote the antioxidant activity, Functional food material for prevention and improvement of dementia, characterized in that it further comprises liver, lotus root and three hundred seconds. Functional food for the prevention and improvement of dementia, comprising the food material of claim 5. delete delete