KR20190125804A - Composition for Preventing or Treating Cognitive Dysfunction Comprising Extraction of Gentiana lutea or Compound Isolated therefrom - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating or treating cognitive impairments or a composition for alleviating cognitive abilities or preventing the deterioration of cognitive abilities by including Gentianalutea extract or an active compound isolated therefrom as an active ingredient. According to an embodiment of the present invention, the gentianalutea extract and the compound significantly inhibit the death of brain nerve cells induced by hydrogen peroxide. The composition including gentianalutea extract or the compound isolated therefrom can be used for the prevention, alleviation or treatment of cognitive impairments caused by the death of brain nerve cells and can also be used to improve cognitive abilities or prevent the deterioration of cognitive abilities.

Description

Composition for Preventing or Treating Cognitive Dysfunction Comprising Extraction of Gentiana lutea or Compound Isolated therefrom}

The present invention relates to a composition for the prevention or treatment of cognitive dysfunction comprising a gentian lutea extract or a compound isolated therefrom as an active ingredient.

According to the 2005 statistics of the future population estimates, Korea has entered an aging society in which the elderly population over 65 years has exceeded 7%, and it is expected to become an aging society in 2018 where the elderly population exceeds 14%. have. In other words, with the development of medical technology and the improvement of living standards, the average life span of humans has almost doubled over the last half century, which has led to a rapid increase in the ratio of the elderly to the total population.

On the other hand, cognitive impairment that loses the ability of cognitive function can be divided into dementia and mild cognitive impairment. Mild cognitive impairment complains of subjective memory impairment or abnormality on objective test, but it is not dementia because it does not interfere with daily life and mental function is maintained normally. Before the onset of mild cognitive impairment, the brain is in the process of killing neurons, but at this time there are no symptoms and it is difficult to distinguish from memory impairment due to normal aging. Mild cognitive impairment is more common than dementia, and 15 to 30% of 60-year-olds may fall within this category and increase with age. The condition of mild cognitive impairment currently means a moderate cognitive impairment between normal and Alzheimer's disease, and about 15% of dementia develops every year. In 2050, the world's dementia patients are expected to surpass 100 million people, 3.1 times higher than in 2013. Korea is expected to become the fastest growing country of dementia patients in the world due to the aging population of over 65%. It is emerging as a major social concern.

As society gradually ages, senile dementia has emerged as the biggest health problem that humankind has to solve in the 21st century. The demand for development of functional substances and foods that can prevent and treat dementia and cognitive dysfunctions is increasing. There is a situation.

Korea Patent Registration No. 10-0858357 (registered September 5, 2008).

It is an object of the present invention to provide a pharmaceutical composition that can more effectively prevent or treat cognitive dysfunction.

Another object of the present invention to provide a nutraceutical composition that can more effectively prevent or improve cognitive dysfunction.

Still another object of the present invention is to provide a nutraceutical composition that can effectively improve cognitive ability or prevent cognitive decline.

In order to achieve the above object, the present invention is Gentiana ( Gentiana) lutea ) provides a pharmaceutical composition for preventing or treating cognitive dysfunction comprising the extract as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating cognitive dysfunction, comprising geniopicroside (Gentiopicroside) as an active ingredient.

In order to achieve the above another object, the present invention is Gentiana ( Gentiana) lutea ) provides a dietary supplement for preventing or improving cognitive dysfunction comprising the extract as an active ingredient.

In another aspect, the present invention provides a health functional food composition for preventing or improving cognitive dysfunction comprising a gentiopicroside (Gentiopicroside) as an active ingredient.

In order to achieve the above another object, the present invention is Gentiana ( Gentiana) lutea ) provides a dietary supplement for improving cognitive abilities or preventing cognitive decline, comprising the extract as an active ingredient.

In addition, the present invention provides a health functional food composition for improving cognitive ability or preventing cognitive decline, comprising genopicroside (Gentiopicroside) as an active ingredient.

The present invention relates to a composition for preventing, improving, or treating cognitive impairment or improving cognitive ability or preventing cognitive deterioration, comprising Gentiana lutea extract or an active compound isolated therefrom as an active ingredient. According to the present invention, the Gentiana extract or the effective compound isolated therefrom has a remarkably excellent activity of protecting brain cells by inhibiting brain cell death, thereby preventing or treating cognitive dysfunction, improving cognitive ability or lowering cognitive ability. It can be usefully used as a composition for preventing.

Figure 1 shows the results of confirming the cell survival rate after treatment with hydrogen peroxide (H ₂ O ₂ ) by concentration in the brain neuron cell line in order to induce brain neuron cell death.
2 is a result confirming the brain neuroprotective activity of the gentian lutea extract.
Figure 3 is a result of confirming the brain neuroprotective activity of the active ingredient gentiopicroside (Gentiopicroside) isolated from gentiana lutea extract.
Figure 4 is a result of comparing the neuroprotective activity of gentian lutea extract and genthiopicroside.
Figure 5 is a result showing the HPLC profile and NMR analysis data of genthiopicroside in the gentian lutea extract.

Hereinafter, the present invention will be described in more detail.

The present invention is Gentiana ( Gentiana) lutea ) provides a pharmaceutical composition for preventing or treating cognitive dysfunction comprising the extract as an active ingredient.

In addition, the present invention is Gentiana ( Gentiana) lutea ) provides a dietary supplement for preventing or improving cognitive dysfunction comprising the extract as an active ingredient.

The gentian lutea extract may be extracted from 20% to 50% ethanol aqueous solution of the roots of gentian lutea, preferably the roots of gentian lutea may be extracted with 30% aqueous ethanol solution. When extracted from, it is preferable because the effect of protecting the brain neurons is maximized.

More preferably, the cognitive dysfunction is dementia, Parkinson's disease, learning disorder, learning, agnosia, amnesia, aphasia, pick disease However, the present invention may be selected from the group consisting of mild cognitive impairment and Binswangers disease, but is not limited thereto.

That is, according to the present invention, the gentian lutea extract may improve cognitive function, and specifically, the gentian lutea extract may improve cognitive function by inhibiting the death of brain neurons. It can be used as a composition that can prevent, ameliorate or treat cognitive dysfunction.

In addition, the present invention provides a health functional food composition for improving cognitive ability or preventing cognitive decline, including Gentiana lutea extract as an active ingredient.

The gentian lutea extract may be extracted from 20% to 50% ethanol aqueous solution of the roots of gentian lutea, preferably the roots of gentian lutea may be extracted with 30% aqueous ethanol solution. When extracted from, it is preferable because the effect of protecting the brain nerve cells to improve the cognitive ability, or prevent the degradation of the cognitive ability is maximized.

Furthermore, according to an embodiment of the present invention, the core active ingredient showing the effect of protecting the brain neurons in the gentian lutea extract was confirmed, and as a result, the genopicroside was identified as the active compound. It was.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating cognitive dysfunction, which contains gentiopicroside as an active ingredient.

In another aspect, the present invention provides a health functional food composition for preventing or improving cognitive dysfunction comprising a gentiopicroside (Gentiopicroside) as an active ingredient.

In one embodiment of the present invention, the genthiopicroside may be isolated from the 20% to 50% ethanol aqueous extract of the gentian lutea root, and in detail, the 30% ethanol aqueous extract of the gentian lutea root. It may be separated, but is not limited thereto.

More specifically, the genthiopicroside may be represented by the following <Formula 1>.

[Formula 1]

In addition, the cognitive dysfunction is dementia, Parkinson's disease, learning disorder (learning disorder), agnosia, amnesia, aphasia, Pick disease, mild cognition It may be selected from the group consisting of mild cognitive impairment and Binswangers disease, but is not limited thereto.

In addition, the present invention provides a health functional food composition for improving cognitive ability or preventing cognitive deterioration comprising geniopicroside (Gentiopicroside) as an active ingredient.

In one embodiment of the present invention, the genthiopicroside may be isolated from the 20% to 50% ethanol aqueous extract of the gentian lutea root, and in detail, the 30% ethanol aqueous extract of the gentian lutea root. It may be separated, but is not limited thereto.

More specifically, the genthiopicroside may be represented by the following <Formula 1>.

[Formula 1]

As used herein, the term "prevention" refers to any action that inhibits or delays a disease by administration of a composition containing the gentian or lutea extract or genthiopicroside. In addition, the term "treatment" used in the present invention means any action that improves or cures the symptoms of the disease by administration of the composition containing the gentian or lutea extract or genthiopicroside.

The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier, excipient or diluent in addition to the above-mentioned active ingredient for administration. The carrier, excipient and diluent may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical compositions of the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, or sterile injectable solutions according to conventional methods. . In detail, when formulated, it may be prepared by using diluents or excipients such as fillers, weights, binders, wetting agents, disintegrating agents, and surfactants which are commonly used. Solid preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. Such solid preparations may be prepared by mixing at least one excipient such as starch, calcium carbonate, sucrose, lactose, gelatin and the like with the gentiana lutea extract or genthiopicroside. In addition to simple excipients, lubricants such as magnesium stearate, talc can also be used. It may be prepared by adding various excipients such as humectants, sweeteners, fragrances, preservatives and the like in addition to liquid oral liquids or liquid paraffin for oral use. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations and tasks. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. As the base of the suppository, utopsol, macrosol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Suitable dosages of the compositions of the present invention vary with the condition and weight of the patient, the extent of the disease, the form of the drug, and the time, but may be appropriately selected by one of ordinary skill in the art, such as the gentiana lutea extract or genthiopicroside. The daily dosage of is preferably 1 mg / kg to 500 mg / kg, may be administered once to several times daily if necessary.

The health functional food composition includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof. , Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. Others may contain pulp for the production of natural fruit juices, synthetic fruit juices and vegetable drinks. These components can be used independently or in combination. In addition, the nutraceutical composition is in the form of any one of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcohol and vitamin complex Can be.

In addition, the health functional food composition may further include a food additive, the suitability as a "food additive" is the item according to the General Regulations and General Test Methods of the Food Additives Code approved by the Food and Drug Administration unless otherwise specified It is determined by the standard and the standard.

Items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, cinnamon acid, natural additives such as color pigments, licorice extract, crystalline cellulose, high-quench pigments, guar gum, L Mixed preparations, such as a sodium glutamate preparation, a noodles addition alkali agent, a preservative preparation, and a tar pigment preparation, etc. are mentioned.

At this time, the gentian or lutea extract or genthiopicroside according to the present invention added to the food in the process of preparing the health functional food composition may be appropriately added or decreased as necessary, preferably 100 parts by weight of food It is preferable to add 1 part by weight to 15 parts by weight.

In addition, the present invention provides a reagent composition that inhibits the death of brain neurons in vitro, including the Gentiana lutea extract as an active ingredient.

The present invention also provides a reagent composition that inhibits the death of brain neurons in vitro, including genthiopicroside as an active ingredient.

Furthermore, the present invention provides a method for inhibiting death of brain neurons, comprising the step of treating a gentian or lutea extract to an animal other than a human.

The present invention also provides a method for inhibiting death of brain neurons, comprising the step of treating a genthiopicroside in an animal other than a human.

Hereinafter, examples will be described in detail to help understand the present invention. However, the following examples are provided to more fully explain the present invention to those skilled in the art, and the scope of the present invention is not limited to the following examples, only to illustrate the contents of the present invention. no.

Example 1 Checking Brain Neuronal Survival Rate During Hydrogen Peroxide Treatment

The inventors of the present invention performed in vitro experiments according to the functional functional food evaluation-cognitive improvement guide of the Ministry of Food and Drug Safety.

First, SH-SY5Y cells (ATCC ^® CRL-2266 ™) and PC12 cells (ATCC ^® CRL-1721 ™), which are brain nerve cell lines, were seeded in 96 well plates, and then cultured at 37 ° C. for 24 hours. Thereafter, in order to induce brain neuronal cell death, hydrogen peroxide (H ₂ O ₂ ) was treated at various concentrations, and cell viability was confirmed through MTT assay.

As a result, as shown in FIG. 1, 50% brain cell death occurred in 100 μM hydrogen peroxide (H ₂ O ₂ ) treatment in SH-SY5Y cells, and 50% in 50 μM hydrogen peroxide treatment in PC12 cells. Brain cell death was induced (indicated by a red box).

Therefore, each concentration of hydrogen peroxide, which causes about 50% cell death for each cell, was selected as the optimal concentration to be used for measuring the neuronal neuroprotective activity of gentiana lutea extract.

Example 2 Brain Neuronal Protective Activity of Gentiana Lutea Extract

Then, in order to confirm the brain cell survival rate and brain cell protection activity of the Gentiana Lutea extract, 50 g of Chinese Gentiana Lutea root was added to 0.5 L of 30% ethanol aqueous solution, and the process was cooled twice for 24 hours. After filtration and concentration under reduced pressure, 2.3 g of Gentiana lutea extract was prepared.

Subsequently, the gentian lutea extract was treated at 2, 10, and 50 µg / ml concentrations and cultured for 24 hours in the SH-SY5Y cells and PC12 cells prepared in Example 1, and then hydrogen peroxide was treated in the cells for 1 hour. As described in Example 1, by measuring the cell viability, the brain cell protective activity of the extract during the induction of brain cell death was measured.

As a result, as shown in Figure 2, it was confirmed that each control neuronal cells are killed by hydrogen peroxide in the control group by about 50%, and the group treated with Gentiana lutea extract was confirmed that the protective effect of the brain cells. In particular, at 10 μg / ml and 50 μg / ml concentrations of Gentiana lutea, remarkably high brain cell protective activity was observed in both SH-SY5Y cells and PC12 cells, which effectively inhibited brain neuronal cell death.

These experimental results confirmed that Gentiana lutea extract has a good effect on improving cognitive function.

Example 3 Confirmation of the Brain Neuronal Protective Activity of Geniopicroside (GPS)

This time, 2.3 g of Gentiana Lutea extract extracted with 30% aqueous ethanol solution was suspended in 25 mL of distilled water in order to confirm the brain neuron protective activity of the active compound isolated from the Gentiana Lutea extract. After chloroform was added, the mixture was repeatedly extracted with shaking, and the aqueous layer was separated by fractionation in a separating funnel. The fractionated aqueous layer was partitioned with ethyl acetate (EtOAc) and butanol (BuOH) in the same manner, and each layer was concentrated under reduced pressure to give 102.1 mg of chloroform layer, 81.1 mg of EtOAc layer, and 452.7 mg of BuOH layer. Subsequently, the material was separated by performing ODS gel chromatography on the obtained BuOH layer. At this time, 6 small fractions (GL-B-01 to 06 fractions) were obtained using a 25% methanol aqueous solution, and GL-B-03 fractions were selected and prep-HPLC was performed (10-40% methanol). As a result, the genthiopicroside was isolated, and it was confirmed that the genthiopicroside was a key component.

In addition, high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS) and NMR analysis were performed on the gentian lutea extract, FIG. 5. As shown in, the genthiopicroside was confirmed to be the highest content of the gentian lutea extract.

Thereafter, the gentiopicroside (Gentiopicroside; GPS) isolated from the gentiana lutea extract was treated with the SH-SY5Y cell line prepared in Example 1 at concentrations of 20, 50 and 100 uM and incubated for 24 hours. The cells were treated with hydrogen peroxide for 1 hour, and the cell viability was measured as described in Example 1 to measure the brain cell protective activity of genthiopicroside upon induction of brain cell death.

As a result, as shown in Figure 3, it was confirmed that each brain nerve cell is about 50% killed by hydrogen peroxide in the control group, it was confirmed that the protective effect of the brain cells in the group treated with genthiopicroside. In particular, significantly higher brain cell protective activity was observed at the concentrations of genthiopicroside 50 and 100 uM, which effectively inhibited brain neuronal cell death.

<Example 4> Comparison of protective effect of the gentian lutea extract and genthiopicroside brain neurons protective activity

Based on the above results, the neuronal cell protective activity of 10 μg / ml and 50 μg / ml concentrations of Gentiana lutea extract and SH-SY5Y cell lines and 100 μM concentration of Genthiopicroside (GPS) For comparison, the brain cell viability obtained in Examples 2 and 3 is shown in one graph and compared.

As a result, as shown in FIG. 4, the genthiopicroside 100 uM concentration treatment group, which is an active ingredient of the extract, was more effective in protecting the brain cells than the gentian lutea extract 10 μg / ml and 50 μg / ml concentration treatment groups. It was.

Having described the specific part of the present invention in detail, it is obvious to those skilled in the art that such a specific description is only a preferred embodiment, thereby not limiting the scope of the present invention. Do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (13)

Gentiana Gentiana Lutea ) pharmaceutical composition for preventing or treating cognitive dysfunction comprising the extract as an active ingredient. The pharmaceutical composition for preventing or treating cognitive dysfunction according to claim 1, wherein the extract of Gentiana Lutea is extracted from 20% to 50% aqueous ethanol solution. The method of claim 1, wherein the cognitive impairment is dementia, Parkinson's disease, learning disorder, agnosia, amnesia, aphasia, pick disease ), Mild cognitive impairment (Bidwangers disease) and mild cognitive impairment (Binswangers disease) characterized in that the pharmaceutical composition for preventing or treating cognitive dysfunction. The pharmaceutical composition for preventing or treating cognitive dysfunction according to claim 1, wherein the gentiana lutea extract improves cognitive function. The pharmaceutical composition for preventing or treating cognitive dysfunction according to claim 4, wherein the gentiana lutea extract improves cognitive function by inhibiting the death of brain neurons. Gentiana Gentiana Lutea ) Health functional food composition for preventing or improving cognitive dysfunction comprising the extract as an active ingredient. Gentiana Gentiana Lutea ) Health functional food composition for improving cognitive ability or preventing cognitive decline comprising the extract as an active ingredient. A pharmaceutical composition for preventing or treating cognitive dysfunction, comprising geniopicroside as an active ingredient. 9. The pharmaceutical composition for preventing or treating cognitive dysfunction according to claim 8, wherein the genthiopicroside is isolated from an extract of 20% to 50% ethanol aqueous solution of the gentian lutea root. The pharmaceutical composition for preventing or treating cognitive dysfunction according to claim 8, wherein the genthiopicroside is represented by the following <Formula 1>.
[Formula 1]

The method of claim 8, wherein the cognitive dysfunction is dementia, Parkinson's disease, learning disorder, agnosia, amnesia, aphasia, Pick disease ), Mild cognitive impairment (Bidwangers disease) and mild cognitive impairment (Binswangers disease) characterized in that the pharmaceutical composition for preventing or treating cognitive dysfunction. Health functional food composition for preventing or improving cognitive dysfunction comprising genthiopicroside (Gentiopicroside) as an active ingredient. Health functional food composition for improving cognitive abilities or preventing cognitive decline, including gentiopicroside (Gentiopicroside) as an active ingredient.

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