KR20200117501A - Composition for improving damages of neuronal cells or inhibiting apoptosis of neuronal cells - Google Patents

Abstract

The present specification relates to a composition containing baicalin, polyozellin or evodiamine. The composition inhibits SHIP2 activity to inhibit hyperphosphorylation of tau protein, thereby exhibiting an effect of alleviating nerve cell damage or inhibiting nerve cell apoptosis. Therefore, the composition of the present invention can exhibit effects of preventing, alleviating or treating secondary diseases which can occur due to damage or death of nerve cells caused by SHIP2 activation, thereby being able to be used as a pharmaceutical or food composition.

Description

Composition for improving damages of neuronal cells or inhibiting apoptosis of neuronal cells}

The present specification relates to a composition comprising baicalin, polyzaline or ebadiamine.

Alzheimer's dementia is a disease in which memory and thinking skills are gradually lost due to neuronal damage. Biomarkers known in the studies to date are proteins that accumulate inside and outside neurons. Amyloid beta aggregates and accumulates outside neurons, and hyperphosphorylated abnormal tau proteins are gradually accumulated inside neurons. It is known that amyloid beta binds to various receptors on the surface of neurons and, in particular, to FcrRIIb receptors, thereby inducing hyperphosphorylation of tau protein inside neurons.

When amyloid beta binds to the receptor, it phosphorylates the part inside the receptor and activates SHIP2 (SH2 domain-containing phosphatidylinositol 5'-phosphatase 2) to proceed with the hyperphosphorylation of tau protein. Conversely, inhibiting the activity of SHIP2 suppresses the hyperphosphorylation of tau protein, and simultaneously returns to normal memory and learning ability.

SHIP2 plays a role in producing PtdIns(3,4)P2 by removing phosphate groups from PtdIns(3,4,5)P3 attached to the cell surface. In particular, in Alzheimer's patients, the metabolism of phosphatadylinositol is normal It is known that there is a difference.

In the present invention, amyloid beta (1-42) binds to the RcrRIIb receptor and activates the receptor, thereby activating SHIP2 phosphatase, and using a SHIP2 inhibitor to reduce memory loss learning ability due to hyperphosphorylation of tau protein due to activation of GSK3beta. It is intended to be applied to dementia treatment through memory recovery and learning ability recovery.

Korean patent publication KR 10-1839721 B1 Korean Patent Publication No. KR 10-1995-0032202 A Korean Patent Publication No. KR 10-2016-0085581 A

It is an object of the present specification to provide a composition exhibiting an effect of improving neuronal damage caused by SHIP2 activation or inhibiting neuronal death.

In order to achieve the above object, the present invention in one aspect, Baikalin (Baicalein) or golden (Scutellaria baicalensis orScutellaria lateriflora) Extract, polyozellin or blackcurrant containing the same (Polyozellus multiplex) Extract, and Evodiamine or a composition comprising at least one selected from the group consisting of plant extracts including Tetradium genus as an active ingredient.

The composition according to an aspect of the present invention exhibits an effect of improving neuronal damage or inhibiting neuronal cell death by inhibiting the hyperphosphorylation of tau protein by inhibiting SHIP2 activity. Therefore, the composition according to one aspect of the present invention can be used as a pharmaceutical or food composition because it can exhibit the effect of preventing, improving or treating secondary diseases that may occur due to damage or death of nerve cells caused by SHIP2 activation. .

1 is a graph showing SHIP2 activity (%) when treated with baicalin.
2 is a graph showing SHIP2 activity (%) when polyzaline was treated.
3 is a graph showing SHIP2 activity (%) when treated with Ebodiamine.
Figure 4 is a graph showing the amount of PIP ₂ (phosphatidylinositol 4,5-biphosphate; phosphatidylinositol 4,5-biphosphate) produced when each 10 μM of baicalin and ebodiamine are treated.

In one aspect, the present invention, Baikalin (Baicalein) or golden ( Scutellaria baicalensis or Scutellaria containing the same) lateriflora ) extract; Polyozellin or blackcurrant ( Polyozellus multiplex ) extract containing the same; And Evodiamine or Tetradium genus plant extract containing the same; It may be related to a composition comprising one or more selected from the group consisting of as an active ingredient.

In one aspect of the present invention, baicalin may have a structure represented by the following formula (1).

[Formula 1]

In one aspect of the present invention, polyzaline may have a structure represented by the following formula (2).

[Formula 2]

In one aspect of the present invention, Ebodiamine may have a structure represented by the following Chemical Formula 3.

[Formula 3]

In one aspect of the present invention, the composition or golden extract according to one aspect of the present invention may consist of only baicalin as an active ingredient.

In one aspect of the present invention, the composition or blackcurrant extract according to one aspect of the present invention may be composed of only polyzaline as an active ingredient.

In one aspect of the present invention, the composition according to one aspect of the present invention or the plant extract of the genus C. may be composed of only Ebodiamine as an active ingredient.

In one aspect of the present invention, the composition may relate to a composition for improving nerve cell damage or inhibiting neuronal cell death due to SHIP2 (SH2 domain-containing phosphatidylinositol 5'-phosphatase 2) activation.

In one aspect of the present invention, the composition may relate to a composition for inhibiting SHIP2 activity in nerve cells.

In one aspect of the present invention, the composition may be a composition for treatment, prevention or improvement of neurodegenerative diseases caused by SHIP2 activation.

In one aspect of the present invention, the neurodegenerative diseases caused by activation of SHIP2 are Alzheimer's disease, frontotemporal lobar degeneration, corticobasal degeneration, and progressive supranuclear palsy. And Pick's disease (Pick's disease) may be at least one selected from the group consisting of.

In one aspect of the present invention, the composition may be a composition for the treatment, prevention or improvement of neurodegenerative diseases caused by PIP ₂ , and at this time, neurodegenerative diseases caused by PIP ₂ produced by activation of SHIP2 are Alzheimer's It may be one or more selected from the group consisting of disease, temporal lobe degeneration, cortical phase degeneration, degenerative supranuclear palsy, and Pick's disease.

In one aspect of the present invention, the composition may be a composition for the treatment, prevention, or improvement of degenerative neurological diseases caused by tau protein hyperphosphorylation, and the neurodegenerative diseases caused by tau protein hyperphosphorylation are Alzheimer's disease, It may be one or more selected from the group consisting of temporal lobe degeneration, cortical phase degeneration, degenerative supranuclear palsy, and Pick's disease.

SHIP2 (SH2 domain-containing phosphatidylinositol 5'-phosphatase 2) of the present invention is PtdIns(3,4,5)P ₃ (Phosphatidylinositol (3,4,5)-trisphosphate, phosphatidylinositol ( PtdIns(3,4)P ₂ (Phosphatidylinositol (3,4)-biphosphate, phosphatidylinositol (3,4)-biphosphate, or PIP ₂ ) by removing the phosphoric acid group from 3,4,5)-triphosphate) As an enzyme (phosphatase) that produces), when beta amyloid (1-42) binds to the FcrRIIb (Fcg-receptor IIb) receptor and activates the receptor, SHIP2 is activated, and GSK-3β Activation causes tau protein to be hyperphosphorylated. Memory loss or learning ability decreases due to the hyperphosphorylation of the tau protein, and further neurodegenerative diseases may occur.

In one aspect of the present invention, the composition may be a pharmaceutical or food composition.

In one aspect of the present invention, the baicalin is golden ( Scutellaria baicalensis or Scutellaria lateriflora ), or polyzaline is isolated or purified from blackcurrant ( Polyozellus multiplex ), or Ebodiamine may be isolated or purified from Tetradium genus plant extract, or Baicalin, polyzaline or ebadiamine may be those that are generally sold.

In one aspect of the present invention, the at least one extract selected from the group consisting of the golden extract, the blackcurrant extract, and the genus plant extract may be one or more extracts selected from the group consisting of water, an organic solvent, and a mixture thereof. Specifically, in one aspect of the present invention, the organic solvent may be one or more selected from the group consisting of C ₁ ~ C ₆ lower alcohol, butylene glycol, and propylene glycol.

In one aspect of the present invention, the at least one extract selected from the group consisting of the golden extract, the blackcurrant extract, and the genus plant extract is one or more organic solvents selected from the group consisting of the golden extract, blackcurrant extract and the genus plant extract It may be a fraction obtained by fractionating the extract into one or more selected from the group consisting of water, organic solvents, and mixtures thereof. Specifically, in one aspect of the present invention, the at least one extract selected from the group consisting of the golden extract, the blackcurrant extract, and the genus plant extract is one or more selected from the group consisting of the golden extract, blackcurrant extract and the genus plant extract The organic solvent extract may be one or more fractions selected from the group consisting of water, ethyl acetate, and mixtures thereof.

In one aspect of the present invention, the gold is gold ( Scutellaria baicalensis or Scutellaria lateriflora ) It may be one or more selected from the group consisting of leaves, flowers, stems, fruits and roots of the herb. Specifically, the gold may be the root of a golden herb.

In one aspect of the present invention, the genus Schizophrenia plant is Schizophrenia ( Tetradium daniellii ) and Sewage Tree ( Tetradium ruticarpum ) may be one or more selected from the group consisting of.

In one aspect of the present invention, the genus Schizophrenia plant is Schizophrenia ( Tetradium daniellii ) and Sewage Tree ( Tetradium ruticarpum ) It may be at least one selected from the group consisting of one or more woody leaves, flowers, stems, fruits, roots, stems or roots of the heartwood selected from the group consisting of these.

In one aspect of the present invention, baicalin may be isolated or purified from gold, and specifically, the gold may be the golden extract or the golden organic solvent extract.

In one aspect of the present invention, baicalin is fractionated by one or more selected from the group consisting of water, ethyl acetate, and mixtures thereof from the golden extract or the golden organic solvent extract, and then the fraction is again water, ethyl acetate, and their It may be fractionated into one or more selected from the group consisting of mixtures and separated by chromatography.

In one aspect of the present invention, polyzaline may be isolated or purified from blackcurrant, and specifically, the blackcurrant may be the blackcurrant extract or the blackcurrant organic solvent extract.

In one aspect of the present invention, polyzaline is fractionated by one or more selected from the group consisting of water, ethyl acetate, and mixtures thereof from the blackcurrant extract or the blackcurrant organic solvent extract, and the fraction is again water, ethyl acetate, and It may be fractionated into one or more selected from the group consisting of a mixture thereof and separated by chromatography.

In one aspect of the present invention, the Ebodiamine may be isolated or purified from a plant of the genus H. genus, and specifically, the plant of genus H. genus may be an extract of the plant of genus hex or an organic solvent extract of a genus plant.

In one aspect of the present invention, the Ebodiamine is fractionated by one or more selected from the group consisting of water, ethyl acetate, and mixtures thereof by fractioning the Ebodiamine plant extract or the organic solvent extract of the plantain genus, and the fraction again water, It may be fractionated into at least one selected from the group consisting of ethyl acetate and mixtures thereof, and separated by chromatography.

In one aspect of the present invention, at least one selected from the group consisting of baicalin, polyzaline, and ebadiamine may have a concentration of 0.01 μM to 10 M based on the total volume of the composition. Specifically, in one aspect of the present invention, at least one selected from the group consisting of baicalin, polyzaline and ebadiamine is 0.01 μM or more, 0.1 μM or more, 0.2 μM or more, 0.3 μM based on the total volume of the composition. Or more, 0.4 μM or more, 0.5 μM or more, 0.7 μM or more, 0.8 μM or more, 0.9 μM or more, 1.0 μM or more, 1.5 μM or more, 2.0 μM or more, 2.5 μM or more, 2.8 μM or more, 3.0 μM or more, 4.0 μM or more, 5.0 μM or more, 6.0 μM or more, 7.0 μM or more, 8.0 μM or more, 8.3 μM or more, 8.5 μM or more, 9.0 μM or more, 10.0 μM or more, 15.0 μM or more, 20.0 μM or more, 25.0 μM or more, 30.0 μM or more, 50.0 μM Or more, 100 μM or more, 1000 μM or more, 10 mM or more, 100 mM or more, or 1 M or more, 10 M or less, 1 M or less, 100 mM or less, 10 mM or less, 1000 μM or less, 100 μM or less, 50.0 μM or less, 30.0 μM Or less, 25.0 μM or less, 20.0 μM or less, 15.0 μM or less, 10.0 μM or less, 9.0 μM or less, 8.5 μM or less, 8.3 μM or less, 8.0 μM or less, 7.0 μM or less, 6.0 μM or less, 5.0 μM or less, 4.0 μM or less, 3.0 μM or less, 2.8 μM or less, 2.5 μM or less, 2.0 μM or less, 1.5 μM or less, 1.0 μM or less, 0.9 μM or less, 0.8 μM or less, 0.7 μM or less, 0.5 μM or less, 0.4 μM or less, 0.3 μM or less, 0.2 μM It may be a concentration of 0.1 μM or less, or 0.01 μM or less.

In one aspect of the present invention, one or more extracts selected from the group consisting of the golden extract, blackcurrant extract, and hemania plant extract may be in a concentration of 0.1 ug/ml to 1000 ug/ml based on the total volume of the composition. . Specifically, in one aspect of the present invention, the one or more extracts selected from the group consisting of the golden extract, blackcurrant extract, and hemanthus plant extract is 0.1 ug/ml or more, 0.5 ug/ml or more, based on the total volume of the composition. , 1 ug/ml or more, 2 ug/ml or more, 3 ug/ml or more, 4 ug/ml or more, 5 ug/ml or more, 5.55 ug/ml or more, 10 ug/ml or more, 15 ug/ml or more, 16 ug/ml or more, 16.5 ug/ml or more, 16.6 ug/ml or more, 16.8 ug/ml or more, 17.0 ug/ml or more, 17.1 ug/ml or more, 17.2 ug/ml or more, 17.5 ug/ml or more, 18 ug/ ml or more, 20 ug/ml or more, 50 ug/ml or more, 100 ug/ml or more, 500 ug/ml or more, or 1000 ug/ml or more and 2000 ug/ml or less, 1000 ug/ml or less, 500 ug/ ml or less, 100 ug/ml or less, 50 ug/ml or less, 20 ug/ml or less, 18 ug/ml or less, 17.5 ug/ml or less, 17.2 ug/ml or less, 17.1 ug/ml or less, 17.0 ug/ml or less , 16.8 ug/ml or less, 16.6 ug/ml or less, 16.5 ug/ml or less, 16 ug/ml or less, 15 ug/ml or less, 10 ug/ml or less, 5.55 ug/ml or less, 5 ug/ml or less, 4 It may be less than ug/ml, less than 3 ug/ml, less than 2 ug/ml, less than 1 ug/ml, less than 0.5 ug/ml, or less than 0.1 ug/ml, but is not limited thereto.

In this specification "golden ( Scutellaria baicalensis or Scutellaria lateriflora )" refers to a plant of the Lamiaceae family of dicotyledons of the genus plant family. It is known to have efficacy against antipyretic, diuretic, branch, otitis, and anti-inflammation, but SHIP2 inhibitory effect or SHIP2 activation. There is no known efficacy in improving cell damage or inhibiting death.

In the present specification, the term " Polyozellus multiplex " refers to a mushroom of the genus Basidiomyces mycosis fungus sub-family Democratic Rum mushroom order Chimney mushroom and Blackcurrant. It is known to be effective in anticancer, prevention of food poisoning, and treatment of diabetes, but there is no known about the efficacy of inhibiting SHIP2 or improving nerve cell damage caused by SHIP2 activation or inhibiting death.

In the present specification, "Tetradium genus plant" refers to a plant of the genus Dicotyledons of the genus Dicotyledonous plant family of the genus S. The plant of the genus Schizophrenia is a shrub ( Tetradium daniellii ), a sallow tree ( Tetradium ruticarpum ), Tetradium austrosinense , Tetradium calcicola calcicola ), Tetradium prascinifolium fraxinifolium ), Tetradium glabrifolium , and Tetradium trichotomium trichotomum ) may be one or more selected from the group consisting of. It is known to have efficacy against tosa-gwakran, headache, toothache, and neuralgia, but there is no known efficacy for inhibiting SHIP2 or improving neuronal damage or suppressing death due to SHIP2 activation.

In the present specification, "extract" includes all substances obtained by extracting the components of natural substances, regardless of the extraction method, extraction solvent, extracted component, or form of the extract, and after extracting the substance obtained by extracting the component of a natural substance, It is a broad concept including all substances that can be obtained by processing or processing by a method, and specifically, the processing or treatment may be additional fermentation or enzyme treatment of the extract. Therefore, in the present specification, the extract is a broad concept including a fermented product, a concentrate, and a dried product, and specifically, in the present specification, the extract may be a fermented product.

In one aspect of the present invention, "golden extract", "blackcurrant extract" or "shrub extract" is regardless of the extraction method, extraction solvent, extracted component or extract form, gold, blackcurrant, or genus It includes all substances obtained by extracting plant components, and includes substances obtained through an extraction method including a process of treatment with heat, acid, base, enzyme, etc. in the process of extracting the components. In addition, it is a broad concept that includes all substances that can be obtained by extracting the components of gold, blackcurrant, or hemania plant and then processing or processing them by other methods after extraction. Specifically, the processing or treatment may be additional fermentation or enzymatic treatment of a golden extract, blackcurrant extract, or an extract of a genus plantain. Therefore, the golden extract, blackcurrant extract, or the genus plant extract of the present specification may be a fermented product.

In one aspect of the invention, "Golden ( Scutellaria baicalensis or Scutellaria lateriflora )" may be in the form of an extract, raw gold, a pulverized product of the herbal medicine itself, a dried product of a herbal medicine, a dried pulverized product of a herbal medicine, or a fermented product of gold, but are not limited thereto. In one aspect, "Blackcurrant ( Polyozellus multiplex )" may be in the form of an extract, or raw blackcurrant, a pulverized product of a herbal medicine itself, a dried product of a herbal medicine, a dried crushed product of a herbal medicine, a fermented product of blackcurrant, In one aspect of the present invention, the "Tetradium genus plant" is in the form of an extract, or a raw genus plant, the pulverized product of the herbal medicine itself, the dried product of the herbal medicine, the drying of the herbal medicine It may be a pulverized product or a fermented product of a genus plant, but is not limited thereto. Also, there is no limitation on the method of obtaining the gold, blackcurrant, or plant genus used in the present specification, and cultivate and use or purchase a commercially available one It may be used, and part or all of the above-ground part or the root part of the golden herb may be used, part or all of the blackcurrant may be used, and part or all of the above-ground part or the root part of the woody plant of the genus C. can be used. Specifically, one or more selected from the group consisting of leaves, flowers, stems, fruits, and roots of golden herbs may be used, and one or more selected from the group consisting of blackcurrant mushroom stalks, mustards, etc. may be used, and At least one selected from the group consisting of woody leaves, flowers, stems, fruits, roots, stems, or roots of the heartwood and these may be used. It is not limited as long as it is a raw material in a form suitable for extracting the active ingredients of each of the golden, blackcurrant and hex.

In one aspect of the present invention, water includes distilled water or purified water, and the organic solvent includes alcohols and acetone, ether, ethyl acetate, diethyl ether, ethyl methyl ketone, and examples of lower alcohols of C ₁ to C ₆ It includes at least one selected from the group consisting of chloroform, but is not limited thereto.

In one aspect of the present invention, the golden extract, the blackcurrant extract, or the genus plant extract may include a C ₁ ~ C ₆ alcohol extract of gold, blackcurrant or hemanis plant, and specifically, the alcohol is methanol or ethanol Can be

In one aspect of the present invention, the golden extract, the blackcurrant extract or the genus plant extract comprises the step of extracting the golden extract, the blackcurrant extract or the genus plant extract with water, an organic solvent, or a mixture thereof It can be obtained by

In one aspect of the present invention, the golden extract, the blackcurrant extract or the genus plant extract may be a crude extract of a solvent selected from the group consisting of water, an organic solvent, and a combination thereof. The organic solvent may be a C ₁ -C ₆ alcohol, specifically, the C ₁ -C ₆ alcohol may be methanol or ethanol. In one aspect of the present invention, when extracting the golden extract, the blackcurrant extract or the genus plant extract with a solvent, it corresponds to about 5 to 15 times (v/w) of the golden extract, the blackcurrant extract or the genus plant extract It is preferable to extract by adding a solvent, specifically, it is preferable to extract by adding about 10 times the solvent, but is not limited thereto.

In one aspect of the present invention, the extraction may be hot water extraction, ethanol extraction, heat extraction, cold needle extraction, reflux extraction, reflux cooling extraction, or ultrasonic extraction, etc., and there is no limitation as long as the extraction method is obvious to those skilled in the art. The extraction may be hot water extraction or ethanol extraction.

In one aspect of the present invention, extraction may be performed at room temperature, but for more efficient extraction, it may be performed under heated conditions, preferably at a temperature of about 40 to 100 °C, more preferably about 65 to 75 °C. It can be extracted from, but is not limited thereto. The extraction time may be performed for about 2 hours to about 48 hours, specifically 18 hours to 36 hours, more specifically 20 hours to 28 hours, most specifically 22 hours to 26 hours, but is not limited thereto, It may vary depending on conditions such as extraction solvent and extraction temperature. The extraction may be extracted once or more several times in order to obtain a larger amount of the active ingredient, preferably 1 to 5 times, more preferably 3 times continuous extraction to use the combined extract.

In one aspect of the present invention, the golden extract, the blackcurrant extract, or the genus plant extract may include a crude extract of a golden extract, a blackcurrant extract, or an extract of a genus Hemiaceae plant as described above, and the crude extract has a low polarity. It can also be included as a soluble fraction of an organic solvent obtained by further extraction with an organic solvent. In one aspect of the present invention, hexane, methylene chloride, ethyl acetate, n-butanol, and the like may be used as the organic solvent, but is not limited thereto. The extract extracted by the above method or a soluble fraction of the extract may be used as it is, but may be used in the form of an extract after filtration and concentration, and may be used as a dried product after concentration.

In one aspect of the present invention, drying may be evaporation drying, spray drying, or freeze drying, and specifically, freeze drying may be performed at -50 to -70°C for 3 to 4 days during freeze drying.

The pharmaceutical composition according to one aspect of the present invention may be in various oral or parenteral dosage forms. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, soft or hard capsules, and the like, and these solid preparations include at least one excipient, such as starch, calcium carbonate, and sucrose, in one or more compounds. Or it is prepared by mixing lactose, gelatin, and the like. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, and syrups.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as humectants, sweeteners, fragrances, and preservatives may be included. have. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.

The pharmaceutical dosage form of the composition according to an aspect of the present invention may be used in the form of a pharmaceutically acceptable salt thereof, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable set. The salt is not particularly limited as long as it is pharmaceutically acceptable, and examples include hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid. , Benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, and the like can be used.

The composition according to an aspect of the present invention may be administered parenterally or orally as desired, and may be administered in an amount of 0.1 to 500 mg per 1 kg of body weight per day, specifically 1 to 100 mg per day. Can be administered separately. The dose administered to a specific patient may vary according to the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, disease severity, and the like.

The pharmaceutical composition according to an aspect of the present invention, respectively, according to a conventional method, oral dosage forms such as powders, granules, tablets, soft or hard capsules, suspensions, emulsions, syrups, drinks, aerosols, ointments, and skin such as creams. It can be formulated and used in any form suitable for pharmaceutical preparations, including external preparations, suppositories, injections, and sterile injectable solutions, and preferably can be formulated and used in the form of injections or skin external preparations.

The composition according to an aspect of the present invention may be administered to mammals such as rats, mice, livestock, humans, etc. by various routes such as parenteral or oral, and all modes of administration can be expected, for example, It can be administered by oral, transdermally, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The composition according to an aspect of the present invention may be administered by various routes that can be easily applied by a person skilled in the art.

In one aspect of the present invention, the food composition may be a health functional food composition.

The formulation of the food composition according to an aspect of the present invention is not particularly limited, but may be formulated as, for example, a tablet, granule, powder, liquid such as a drink, caramel, gel, bar, or the like. In the food composition of each formulation, in addition to the active ingredients, ingredients commonly used in the field may be appropriately selected and blended by a person skilled in the art according to the formulation or purpose of use without difficulty, and synergistic effects may occur when applied simultaneously with other ingredients.

In the food composition according to an aspect of the present invention, the determination of the dosage of the active ingredient is within the level of those skilled in the art, and the daily dosage thereof is, for example, 0.1 mg/kg/day to 5000 mg/kg/day, more Specifically, it may be 50 mg/kg/day to 500 mg/kg/day, but is not limited thereto, and may vary depending on various factors such as age, health condition, and complications of the subject to be administered.

Food compositions according to one aspect of the present invention include, for example, various foods such as chewing gum, caramel products, candies, frozen desserts, and confectionery, beverage products such as soft drinks, mineral water, alcoholic beverages, and health including vitamins or minerals. It may be a functional food.

In addition to the above, the food composition according to an aspect of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and enhancing agents (cheese, chocolate, etc.), pectic acid and salts thereof. , Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like. In addition, functional food compositions according to an aspect of the present invention may include flesh for the manufacture of natural fruit juice and fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The proportion of these additives is not so critical, but is generally included in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition herein.

Hereinafter, the configuration and effects of the present specification will be described in more detail with reference to Examples and Test Examples. However, these Examples and Test Examples are provided for illustrative purposes only to aid understanding of the present specification, and the scope and scope of the present specification are not limited by the following examples.

[ Example ] Baikalin , Polycrop , Ebodiamine Preparations

Baicalin (Example 1), polyzaline (Example 2), and Ebodiamine (Example 3) used as active ingredients in the present invention were each purchased from Sigma-Aldrich, USA. . The purchased sample was made into a 10 mM DMSO solution, diluted 3 times during the experiment, and used in the experiment.

[Experimental Example 1] Confirmation of SHIP2 activity inhibition effect

Using the Malachite green assay in 384 well plate (Corning 3640, NY, USA), the effect of inhibiting SHIP2 phosphatase activity by the three active ingredients of Examples 1 to 3 was confirmed.

SHIP2 (SH2 domain-containing phosphatidylinositol 5'-phosphatase 2) phosphatase and Ptdlns(3,4,5)P ₃ were mixed with each of the samples of Examples 1 to 3 to make 15 μL. Specifically, PtdIns(3,4,5)P ₃ (Echelon P-3908, Salt lake city, UT, USA) was dissolved in distilled water and used as 1 mM stock, and 100 μM was used in the experiment. The SHIP2 phosphatase was diluted with a reaction buffer (6mM MgCl ₂ , 10mM HEPES, 0.25mM EDTA, 0.1% CHAPS, 250mM sucrose, dH2O, pH±7.25) to make 30nM during the reaction. Reaction buffer was prepared and used on the same day, and must be kept on ice when using. The enzyme reaction was carried out as follows. First, SHIP2 phosphatase and the sample were incubated at 27°C for 20 minutes. Thereafter, PtdIns(3,4,5)P ₃ was added and incubated again for 50 minutes at the same temperature for 50 minutes. After the incubation was completed, 40 μL of Malachite green solution (Echelon K-1501, Salt lake city, UT, USA) was added and incubated at 27° C. for 20 minutes. After the reaction was completed, the 384 well plate was measured for absorbance at 620 nm using a Spectra Max M2 (molecular Devices, USA) instrument. IC ₅₀ (the concentration that inhibits 50% SHIP2 activity) was calculated using GraphPad Prism 5.0 (San Diego, CA), and the results are shown in FIGS. 1 to 3. FIG. 1 shows the inhibition rates of SHIP2 activity of baicalin of Example 1, polysarin of Example 2, and Ebodiamine of Example 3, respectively.

As shown in FIGS. 1 to 3, when the baicalin concentration was 2.05 uM, the polyzaline concentration was 2.4 uM, and the eboiamine concentration was 8.8 uM, the activity of SHIP2 decreased to 50%. That is, baicalin and polyzaline showed an IC ₅₀ value equivalent to that of the positive control group (AS90) (Figs. 1 and 2), and baicalin, polyzaline, and ebodiamine have the effect of inhibiting SHIP2 activity even at very low concentrations. I could see that there was.

[Experimental Example 2] PIP ₂ Confirmation of production inhibition effect

In order to confirm the inhibitory effect of PIP ₂ production by the _two active ingredients of Examples 1 and 3, first, 1 x 10 ⁶ HT22 cells induced from the hippocampal neuron of the mouse were placed in a 10 cm cell culture dish. Incubated for 24 hours at 37°C for 24 hours and in a 5% CO ₂ incubator for 24 hours. Thereafter, the culture was performed for 5 hours by exchange with a medium without bovine fetal serum, and then 10 uM of each of the samples of Examples 1 and 3 and 5 uM of VO-Ohpic trihydrate (PTEN inhibitor) were treated on the cells and incubated for 30 minutes. Then, after treatment with 20 mM H ₂ O ₂ for 5 minutes, the cells were harvested with cold 0.5 M TCA (Trichloroacetic acid) and washed with 5% TCA/1 mM EDTA solution. After removing neutral lipids with 3mL of MeOH:CHCl ₃ (2:1), the pellet was vortexed for 30 minutes in 2.25mL of MeOH:CHCl ₃ :conc.HCl (80:40:1) solution and then 3000rpm for 5 minutes. Centrifuged at. Thereafter, the organic layer was collected, dried under reduced pressure, and quantified with an ELISA kit (Echelon Inc. Cat No: K3800), and the results are shown in FIG. 4. DMSO (unstimulated) of FIG. 4 is a control group that is not treated with hydrogen peroxide (H ₂ O ₂ ), and DMSO is a control group that stimulates cells by treatment with hydrogen peroxide.

As shown in FIG. 4, the amount of PIP ₂ produced in the cells stimulated by hydrogen peroxide treatment increased, but the amount of PIP ₂ produced decreased when each treatment with baicalin and ebadiamine decreased, whereas baicalin and ebadiamine produced PIP ₂ It was found that the effect of suppressing the was excellent.

An example of the formulation of the composition according to an aspect of the present invention is described below, but it can be applied to various other formulations, and this is not intended to limit the present specification, but is intended to be described in detail.

[Formulation Example 1] Soft capsule

Baicalin 8mg, vitamin E 9mg, vitamin C 9mg, palm oil 2mg, hydrogenated vegetable oil 8mg, sulfur wax 4mg and lecithin 9mg are mixed and mixed according to a conventional method to prepare a soft capsule filling solution. Each capsule is filled with 400 mg to prepare a soft capsule. In addition to the above, a soft capsule sheet was prepared in a ratio of 66 parts by weight of gelatin, 24 parts by weight of glycerin, and 10 parts by weight of sorbitol liquid, and the filling solution was filled to prepare a soft capsule containing 400 mg of the composition according to the present specification.

[Formulation Example 2] Tablet

Baicalin 8mg, vitamin E 9mg, vitamin C 9mg, galactooligosaccharide 200mg, lactose 60mg and maltose 140mg were mixed, granulated using a fluid bed dryer, and then sugar ester 6mg was added. 500 mg of these compositions are tableted by a conventional method to prepare tablets.

[Formulation Example 3] Drink agent

After mixing 8 mg of baicalin, 9 mg of vitamin E, 9 mg of vitamin C, 10 g of glucose, 0.6 g of citric acid, and 25 g of liquid oligosaccharide, 300 ml of purified water is added and each bottle is filled with 200 ml each. After filling the bottle, sterilize at 130°C for 4-5 seconds to prepare a drink.

[Formulation Example 4] Granules

Baicalin 8mg, vitamin E 9mg, vitamin C 9mg, anhydrous crystalline glucose 250mg and starch 550mg are mixed, molded into granules using a fluid bed granulator, and then filled into a bag to prepare granules.

[Formulation Example 5] Injection

According to the composition shown in Table 1 below, an injection was prepared by a conventional method.

Ingredients content Baikalin 10-50 mg Sterile distilled water for injection Appropriate amount pH adjuster Appropriate amount

[Formulation Example 6] Health Functional Food

Health functional foods were prepared in a conventional manner according to the composition shown in Table 2 below.

Ingredients content Baikalin 20mg Vitamin A acetate 70μg Vitamin E 1.0mg Vitamin B1 0.13mg Vitamin B2 0.15mg Vitamin B6 0.5mg Vitamin B12 0.2μg Vitamin C 10mg Biotin 10μg Nicotinic acid amide 1.7mg Folic acid 50μg Calcium pantothenate 0.5mg Ferrous sulfate 1.75mg Zinc oxide 0.82mg Magnesium carbonate 25.3mg Potassium Phosphate Monobasic 15mg Dicalcium phosphate 55mg Potassium citrate 90mg Calcium carbonate 100mg Magnesium chloride 24.8mg

The composition ratio of the vitamin and mineral mixture is a mixture of components suitable for a health functional food in a preferred embodiment, but the mixing ratio may be arbitrarily modified.

[Formulation Example 7] Healthy Drink

A health drink was prepared by a conventional method according to the composition shown in Table 3 below.

Ingredients content Baikalin 1000mg Citric acid 1000mg oligosaccharide 100 g Taurine 1g Purified water Balance

After mixing the above ingredients according to a conventional health drink manufacturing method, stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and sterilized.

Claims (13)

Baikalin (Baicalein) or gold containing it ( Scutellaria baicalensis or Scutellaria lateriflora ) extract;
Polyozellin or blackcurrant ( Polyozellus multiplex ) extract containing the same; And
Evodiamine or a plant extract of Tetradium genus containing the same; SHIP2 (SH2 domain-containing phosphatidylinositol 5'-phosphatase 2) activation containing at least one selected from the group consisting of as an active ingredient Composition for improving damage or inhibiting neuronal cell death.
Baikalin (Baicalein) or gold containing it ( Scutellaria baicalensis or Scutellaria lateriflora ) extract;
Polyozellin or blackcurrant ( Polyozellus multiplex ) extract containing the same; And
Inhibition of SHIP2 (SH2 domain-containing phosphatidylinositol 5'-phosphatase 2) activity in neurons containing at least one selected from the group consisting of Evodiamine or Tetradium genus plant extract containing the same as an active ingredient Dragon composition.
Baikalin (Baicalein) or gold containing it ( Scutellaria baicalensis or Scutellaria lateriflora ) extract;
Polyozellin or blackcurrant ( Polyozellus multiplex ) extract containing the same; And
Evodiamine or a plant extract of Tetradium genus containing the same; caused by the activation of SHIP2 (SH2 domain-containing phosphatidylinositol 5'-phosphatase 2) containing at least one selected from the group consisting of as an active ingredient. Composition for the treatment, prevention or improvement of neurodegenerative diseases.
The method of claim 3,
The neurodegenerative diseases caused by the activation of SHIP2 include Alzheimer's disease, frontotemporal lobar degeneration, corticobasal degeneration, progressive supranuclear palsy, and Pick's disease. A composition that is at least one selected from the group.
The method according to any one of claims 1 to 4,
Baikalin is golden ( Scutellaria baicalensis or Scutellaria lateriflora ) isolated or purified, or
Polyzaline is isolated or purified from Blackcurrant ( Polyozellus multiplex ), or
Ebodiamine is a composition that is isolated or purified from a plant extract of Tetradium genus.
The method of claim 5,
The gold is golden ( Scutellaria baicalensis or Scutellaria lateriflora ) Herbal leaves, flowers, stems, fruits and roots of at least one selected from the group consisting of, the composition.
The method of claim 6,
The gold is the root of the golden herb, composition.
The method of claim 5,
The genus plant is Tetradium daniellii ), Tetradium ruticarpum , Tetradium ostrosinens austrosinense , Tetradium calcicola , Tetradium fraxinifolium , Tetradium glibrifolium glabrifolium ) and tetradium trichotomum ( Tetradium trichotomum ) is at least one selected from the group consisting of, the composition.
The method of claim 8,
The genus plant is Tetradium daniellii ), Tetradium ruticarpum , Tetradium ostrosinens austrosinense , Tetradium calcicola , Tetradium fraxinifolium , Tetradium glibrifolium glabrifolium ) and tetradium trichotomum ( Tetradium trichotomum ) of at least one tree selected from the group consisting of leaves, flowers, stems, fruits, roots, stems or roots of the heartwood and at least one selected from the group consisting of these.
The method according to any one of claims 1 to 4,
At least one extract selected from the group consisting of the golden extract, the blackcurrant extract and the Hewberry extract is one or more extracts selected from the group consisting of water, organic solvents, and mixtures thereof.
The composition of claim 10, wherein the organic solvent is at least one selected from the group consisting of C ₁ to C ₆ lower alcohol, butylene glycol, and propylene glycol.
The method according to any one of claims 1 to 4,
At least one selected from the group consisting of baicalin, polyzaline, and eboiamine is in a concentration of 0.01 μM to 10 M based on the total volume of the composition.
The method according to any one of claims 1 to 4,
The composition is a pharmaceutical or food composition, composition.

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