KR101910047B1 - Composition for protecting cells from oxidative stress comprising latifolin - Google Patents

Abstract

The present invention relates to a composition comprising latiphorin as an active ingredient. The composition of the present invention protects nerve cells from oxidative stress and inhibits the death of nerve cells. In addition, the expression of antioxidant enzymes that inhibit apoptosis caused by an increase in oxidative stress in cells can be increased. Accordingly, the composition of the present invention can exhibit an effect of preventing, ameliorating or treating a secondary disease that may be caused by the death of nerve cells from oxidative stress, and thus can be used as a pharmaceutical or food composition.

Description

Technical Field [0001] The present invention relates to a composition for protecting cells from oxidative stress including latifolin,

The present disclosure relates to compositions comprising latiphorin.

Oxidative stress in neurons induces many neurological diseases, especially neuropathy such as paralysis, amyotrophic lateral sclerosis (Lou Gehrig's disease), Parkinson's disease, and glutamate toxicity is associated with acute and chronic toxicity in neurons (Andersen, JK et al., Nat. Rev. Neurosci., 2004, 5, S18-S25, Coyle, J. et al. Science 1993, 262, 689-695). Thus, treatment of high concentrations of glutamate in neurons inhibits the influx of cysteine, deficiency of glutathione and increase of active oxygen, resulting in the death of nerve cells. Compounds that inhibit this phenomenon reduce nerve damage by oxidative stress Or as a drug for preventing or preventing cancer.

Latifolin is known as a strong Studies on the pharmacological activity of latifolin as a phenolic compound isolated from odorifera T. Chen have been limited and there has been no report on the protective action of latifurin against nerve damage due to oxidative stress .

The present invention also relates to a pharmaceutical composition for preventing or treating osteoporosis, which comprises a tough extract, a health composition (Korean Patent Laid-Open No. 10-2001-0030064), and an external preparation for skin containing a tall fragrance extract (Korean Patent Publication No. 10-2010-0011764) Has been known.

Korean Patent Publication No. 10-2001-0030064 Korean Patent Publication No. 10-2010-0011764

Andersen, J. K. et al. Nat. Rev. Neurosci. 2004, 5, S18-S25, Coyle, J. et al. Science 1993, 262, 689-695 Yunyi Kang et al., Cellular protection using Flt3 and PI3Ka inhibitors demonstrate multiple mechanisms of oxidative glutamate toxicity, NATURE COMMUNICATIONS, 5: 3672, DOI: 10.1038 / ncomms4672 Nailya S Gliyazova et al., A novel phenoxy thiophene sulphonamide molecule protects against glutamate evoked oxidative injury in a neuronal cell model, BMC Neuroscience 2013, 14: 93 Masayuki Fukui et al., Mechanism of glutamate-induced neurotoxicity in HT22 mouse hippocampal cells, European Journal of Pharmacology, 617 (2009), 1-11

It is an object of the present invention to provide a composition exhibiting an effect of protecting nerve cells against oxidative stress.

In one aspect, the present invention provides a composition comprising latiphorin as an active ingredient.

The composition according to one aspect of the present invention protects nerve cells from oxidative stress and has an effect of inhibiting the death of nerve cells and is useful as an agent for inhibiting apoptosis caused by an increase in oxidative stress in nerve cells And exhibits an effect of increasing expression. Therefore, the composition according to one aspect of the present invention can be used as a pharmaceutical or food composition since it can exhibit the effect of preventing, ameliorating or treating a secondary disease that may be caused by the death of nerve cells from oxidative stress.

1 is earthquake direction (Dalbergia odorifera T. Chen ). < / RTI >
Fig. 2 is a ¹ H NMR spectrum of lattifolines separated in a strong direction.
3 is a < ¹³ > C NMR spectrum of latifolin separated in a strong gradient.
FIG. 4 is a graph showing the cell survival rate of neuronal cells treated with latifolin isolated from strong-smelling fractions.
FIG. 5 is an image photograph showing the survival of nerve cells when cells treated with latifolin isolated from a strong direction. In FIG. 5, the scale bar indicated by white bar is 100 um.

In one aspect, the present invention may relate to a composition comprising latifolin or Dalbergia odorifera T. Chen extract containing the same as an active ingredient.

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

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

In one aspect of the present invention, the composition may be a composition for protecting cells against oxidative stress or a composition for inhibiting apoptosis. Specifically, in one aspect of the present invention, the cell may be a neuron.

In one aspect of the invention, the oxidative stress may be caused by glutamate.

In one aspect of the present invention, the composition may be a composition for treating, preventing or ameliorating a disease caused by the death of nerve cells.

In one aspect of the present invention, the disease caused by nerve cell death may be at least one selected from the group consisting of nerve palsy such as stroke, amyotrophic lateral sclerosis (Lou Gehrig's disease) and Parkinson's disease.

In one aspect of the present invention, the composition may be a composition for treating, preventing or ameliorating a neurodegenerative disease.

In one aspect of the present invention, the neurodegenerative disease may be at least one selected from the group consisting of nerve palsy such as stroke, amyotrophic lateral sclerosis (Lou Gehrig's disease) and Parkinson's disease.

In one aspect of the present invention, the composition may be a composition for treating, preventing or ameliorating a neurological disease caused by oxidative stress, wherein the neurological disease caused by oxidative stress is a neuropathy such as a stroke, Cirrhosis (scleroderma) and Parkinson ' s disease.

In one aspect of the present invention, the composition may be a composition for promoting the expression of an antioxidant enzyme in neurons. In one aspect of the invention, the antioxidant enzyme may be one that inhibits nerve cell death resulting from increased oxidative stress.

In one aspect of the present invention, the composition may be a composition for reducing or inhibiting active oxygen. In one aspect of the present invention, the composition may be a composition for reducing or inhibiting active oxygen generated in a cell, specifically, a neuron.

In one aspect of the present invention, the strong-flavored extract may be one or more extracts selected from the group consisting of water, an organic solvent, and mixtures thereof. Specifically, in one aspect of the present invention, the organic solvent may be at least one selected from the group consisting of C ₁ -C ₆ lower alcohols, butylene glycol, propylene glycol, methylene chloride, and hexane-ethyl acetate mixed solvent, More specifically, the lower alcohol may be ethanol.

In one aspect of the present invention, the strong-flavored extract may be a strong-flavored ethanol extract.

In one aspect of the present invention, the fragrance extract may be a fraction obtained by fractionating the fragrant ethanol extract with a mixture solvent of methylene chloride and hexane-ethyl acetate. Specifically, the volume ratio (v / v) of hexane: ethyl acetate in the hexane-ethyl acetate mixed solvent may be 3-1: 1 to 9, more specifically 1: 3.

In one aspect of the present invention, the earthquake may be at least one selected from the group consisting of wood of leaves, flowers, stems, fruits, roots, roots, or roots of woody logs ( Acronychia pedunculata (L.) Miquel. . Specifically, the tsunamis may be the core of a stem or root of a hardwood tree.

In one aspect of the present invention, the latipurin may be one separated or purified from a strong-smelling flavor, and in particular, the strong-smelling flavor may be the strong-flavored extract or the strong-flavored ethanol extract.

In one aspect of the present invention, latifolin may be obtained by fractionating a strong-flavored or hard-boiled ethanol extract with methylene chloride, fractionating the methylene chloride fraction with a hexane-ethyl acetate mixed solvent, and separating the fraction by chromatography .

In one aspect of the present invention, latifolin can be separated or purified from a strong-shear by a method comprising the steps of:

1) Suspending the extract of Ganoderma lucidum in water and adding methylene chloride to obtain a methylene chloride solvent fraction containing latifolin;

2) adding the methylene chloride solvent fraction to a hexane-ethyl acetate mixed solvent to obtain a hexane-ethyl acetate fraction;

3) separating or purifying the fraction by chromatography.

In the process according to one aspect of the present invention, the volume ratio (v / v) of methylene chloride added for water and fraction to suspend the strongly flavored extract may be 0.5-1.5: 1.5-2.5.

In the method according to one aspect of the present invention, the volume ratio (v / v) of hexane and ethyl acetate in the hexane-ethyl acetate mixed solvent may be 3-1: 1 to 9, 1, 1: 3, 1: 6, or 1: 9, and more specifically 1: 3.

In the method according to one aspect of the present invention, the chromatography may be high performance liquid chromatography (HPLC).

In the process according to one aspect of the present invention, separation or purification by chromatography may be carried out in a volume ratio of acetonitrile: water of 4: 6 to 9: 1, increasing the volume ratio of acetonitrile to 40 minutes to 1 hour and 20 minutes, For 1 hour, wherein separation or purification can be carried out using a reversed phase column.

In one aspect of the present invention, the strong-flavored fragrance extract is prepared by adding water or an organic solvent to the strong flavor and reflux-extraction for 2 to 4 hours, preferably 3 hours; And then drying the extract under reduced pressure.

In one aspect of the invention, the latipurine may be at a concentration of 0.01 μM to 1 M based on the total volume of the composition. Specifically, in one aspect of the present invention, latiphorin is present in an amount of at least 0.01 μM, at least 0.1 μM, at least 0.2 μM, at least 0.3 μM, at least 0.4 μM, at least 0.5 μM, at least 0.7 μM, at least 0.8 μM Or more, at least 0.9 μM, at least 1.0 μM, at least 1.5 μM, at least 2.0 μM, at least 2.5 μM, at least 2.8 μM, at least 3.0 μM, at least 4.0 μM, at least 5.0 μM, at least 6.0 μM, at least 7.0 μM, At least 10 μM, at least 15 μM, at least 20 μM, at least 25 μM, at least 30 μM, at least 50 μM, at least 100 μM, at least 1000 μM, at least 10 mM, at least 100 mM, Or less, 10 mM or less, 1M 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, No more than 8.3 μM, no more than 8.0 μM, no more than 7.0 μM, no more than 6.0 μM, no more than 5.0 μM, no more than 4.0 μM, no more than 3.0 μM, no more than 2.8 μM, no more than 2.5 μM, no more than 2.0 μM, no more than 1.5 μM, no more than 1.0 μM, 0.9 mu M or less, 0.8 mu M or less, 0.7 mu M And, it may be less than 0.5μM, 0.4μM or less, less than 0.3μM, 0.2μM or less, 0.1μM or less, or a concentration of less than 0.01μM.

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

The pharmaceutical composition according to one aspect of the present invention may be various oral or parenteral formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, soft or hard capsules, etc. These solid preparations may contain one or more excipients such as starch, calcium carbonate, sucrose, Or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The pharmaceutical dosage forms of the compositions according to one aspect of the present invention may be used in the form of their pharmaceutically acceptable salts and may also 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 so long as it is pharmaceutically acceptable and includes, for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, , Benzenesulfonic acid, toluenesulfonic acid, and naphthalenesulfonic acid.

The composition according to one aspect of the present invention may be administered parenterally or orally, and may be administered in an amount of 0.1 to 500 mg, preferably 1 to 100 mg per kg of body weight per day, Can be administered separately. The dosage for a particular patient may vary depending on the patient's body weight, age, sex, health condition, diet, time of administration, administration method, excretion rate, severity of disease, and the like.

The pharmaceutical compositions according to one aspect of the present invention may be formulated into tablets, capsules, tablets, pills, tablets, soft or hard capsules, oral formulations such as suspensions, emulsions, syrups, The pharmaceutical composition may be formulated in any form suitable for pharmaceutical preparations including external preparations, suppositories, injectable solutions and sterilized injection solutions, and may be formulated in the form of an injectable or external preparation for skin.

The composition according to one aspect of the present invention can be administered to mammals such as rats, mice, livestock, humans, and the like by various routes such as parenteral, oral, etc. All the ways of administration can be expected, May be administered by oral, transdermal, rectal or intravenous, intramuscular, subcutaneous, intramural or intracerebroventricular injection.

The composition according to one aspect of the present invention may be administered by a variety of routes that are readily applicable to those of ordinary skill in the art. In particular, the pharmaceutical composition according to the present invention can be administered by a route applied to the skin surface as an external preparation for skin.

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

The formulation of the food composition according to one aspect of the present invention is not particularly limited, but may be formulated into, for example, tablets, granules, powders, liquid preparations such as drinks, caramels, gels, bars and the like. The food composition of each formulation can be blended with the ingredients commonly used in the field in addition to the active ingredient without difficulty by those skilled in the art depending on the purpose of formulation or use, and synergistic effect can be obtained when the composition is applied simultaneously with other ingredients.

In the food composition according to one 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 its daily dose is, for example, from 0.1 mg / kg / day to 5000 mg / kg / 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 the age, health condition, and complication of the subject.

The food composition according to one aspect of the present invention can be used in various foods such as chewing gum, caramel product, candy, ice cream, confectionery, beverage such as soft drink, mineral water, alcoholic beverage, health including vitamins and minerals It may be a functional food.

In the present specification, latifulin is a compound having the structure of the above formula (1), specifically, R - (-) - Latipoline, which has a CAS number of 10154-42-4 and a molecular weight of about 286 Da Lt; / RTI >

In the present specification, the strong-feeling fragrance is a medicinal herb which is also called a river or a strong-gauge, and may correspond to a core portion of a stem or root of a strong-hemlock tree. This kind of strong gentle incense helps to stop the bleeding, and it has the efficacy of painfulness. It is a medicinal medicine that treats back and leg pain symptoms due to bleeding, bruise, swelling, Lt; / RTI >

As used herein, the term " extract " includes all substances obtained by extracting components of a natural product, regardless of the extraction method, extraction solvent, extracted component or extract form, Or a substance that can be obtained by processing or treating the extract, in particular, the processing or treatment may be an additional fermentation or enzymatic treatment of the extract. Thus, the extract herein is a broad concept including fermentation products, concentrates, and dried products. Specifically, the extract may be a fermentation product in this specification.

In one aspect of the present invention, " Tropical Fragrant Extract " is intended to encompass all of the substances obtained by extracting a strong-flavored component, regardless of the extraction method, extraction solvent or extracted component or extract form, It contains the substance obtained by the extraction method including the process of treating with heat, acid, base, enzyme, etc., and extracts the substance obtained by extracting strong fragrance ingredient, It is a broad concept that includes all substances that can be obtained by treatment. Specifically, the processing or treatment may be a fermentation, an enzyme treatment or the like in addition to the fragrance extract. Thus, the fragrance extract of the present specification may be a fermented product.

In one aspect of the present invention, " toughness fragrance " may be in the form of an extract, a raw seaweed flavor, a pulverized product of the herbal medicine itself, a dry product of the herbal medicine, a dry pulverized product of the herbal medicine, But is not limited thereto. In addition, there is no limitation on the method of obtaining the strong-smelling fragrance used in the present invention. The fragrant fragrance used in the present invention is not limited to the method of obtaining the fragrance, and may be cultivated or used commercially. More specifically, at least one selected from the group consisting of a hardwood tree, a flower, a stem, a fruit, a core of a root, a stem or a root, and a group consisting of these may be used. In the case of the toughness of the present specification, it is not necessarily manufactured through drying, and it is not limited as long as it is a raw material in a form suitable for extracting the strong active ingredient.

In one aspect of the present invention, the water comprises distilled or purified water, and the organic solvent is selected from the group consisting of alcohols such as C ₁ -C ₅ lower alcohols, acetone, ether, ethyl acetate, diethyl ether, ethyl methyl ketone, Chloroform, and the like, but are not limited thereto.

In one aspect of the present invention, extraction may be performed by hot water extraction, ethanol extraction, heat extraction, cold extraction, reflux extraction, reflux cooling extraction, ultrasonic extraction, etc. Any extraction method obvious to a person skilled in the art is not limited, The extraction may be hot water extraction or ethanol extraction.

In one aspect of the invention, the extraction may be carried out at room temperature, but may be carried out under heating conditions for more efficient extraction, preferably at a temperature of about 40 to 100, more preferably about 65 to 75 But is not limited thereto. The extraction time may be from about 2 to about 48 hours, specifically from about 18 to about 36 hours, more specifically from about 20 to about 28 hours, and most specifically from about 22 to about 26 hours, Solvent and extraction temperature, and the like. The extraction may be carried out one or more times several times to obtain a larger amount of the active ingredient, preferably 1 to 5 times, more preferably 3 times of continuous extraction.

In one aspect of the present invention, the drying may be vacuum drying, evaporation drying, spray drying, or freeze-drying. Specifically, the freeze-drying may be performed at -50 to -70 for 3 to 4 days.

Hereinafter, the configurations and effects of the present invention 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 in order to facilitate understanding of the present specification, and the scope and range of the present specification are not limited by the following examples.

[Example 1] Separation of Latipolin from Gangjin Incense Ethanol Extract

1.2 g of ethanol was added to 600 g of dried and cut herb medicinal herb Gangjin fragrance (core material part of the stem or root of Gangjin japonica) purchased from Kyungdong market, and the mixture was refluxed for 3 hours. The extract was dried under reduced pressure to obtain 82 g of a Gangjin fragrance ethanol extract . In the following experiment, the ethanol extract of Gangjin extract was obtained from the natural materials bank of Korea Oriental Industry Promotion Agency.

11 g of a methylene chloride (dichloromethane) solvent fraction containing latifolin was obtained by suspending the crude extract in ethanol (1 liter) and extracting (fractionating) the same amount of methylene chloride (dichloromethane) twice Respectively.

Then, 11 g of the methylene chloride fraction was eluted with 1 L of a mixed solvent of hexane-ethyl acetate (hexane: ethyl acetate mixing ratio of 3: 1, 1: 1, 1: 3, 1: 6 and 1: 9, respectively) Five fractions were obtained. 3 g of the fraction of the eluent using a mixed solvent in which hexane: ethyl acetate was mixed at a mixing ratio of 1: 3 was increased in volume ratio of acetonitrile to water at a ratio of 4: 6 to 9: 1 for 1 hour , And 76 mg of latifolin was separated and purified in the form of pale yellow powder by high performance liquid chromatography separation using a C18 reverse phase column.

Separation of Latofolin was measured by ¹ H NMR (Fig. 2) and ¹³ C NMR (Fig. 3) in the natural materials bank of the Korean Oriental Industry Promotion Agency and was compared with the existing literature (J. Agric. Food. Chem. 2009, 57, 5707- 5712), and the structure thereof was identified as shown in Fig. The results of ¹ H NMR and ¹³ C NMR spectra measured for separated latifolin are shown in FIGS. 2 and 3, respectively, and the numerical values of the signals are as follows.

^{1 H NMR (500 MHz, CD} 3 OD) δ 6.85-7.19 (4H, m), 6.78 (1H, s), 6.52 (1H, s), 6.15 (1H, ddd, J = 17.0, 10.0, 6.0 Hz) (1H, br d, 6.0), 5.09 (1H, br dd, J = 10.0, 1.5 Hz), 4.72 (1H, br dd, J = , s).

¹³ C NMR (125 MHz, CD ₃ OD)? 154.7, 150.8, 146.2, 140.5, 139.6, 129.6, 129.0, 126.7, 124.1, 118.8, 116.2, 114.7, 114.0, 98.6, 56.2, 55.4, 40.1.

[Test Example 1] Measurement of cell survival rate of neurons by oxidative stress

HT-22 cells derived from mouse hippocampal neurons (Sark Research, USA) were inoculated into DMEM (life technology, 11965-092) medium with 10% fetal bovine serum (Gibco) The cells were subcultured with medium supplemented with 1% penicillin-streptomycin every 3 days while changing medium. In a 96-well plate, 100 uL per well was added to each well. 5,000 HT-22 cells were injected into each well. The cells were cultured in an incubator with 5% CO ₂ at 37 ° C for 24 hours and then dissolved in DMSO at a concentration of 10 mM Latipurin was treated with the concentrations (0.1 μM, 0.3 μM, 0.9 μM, 2.8 μM, 8.3 μM, and 25.0 μM) shown in FIG. After 2 hours of treatment, 10 mM glutamate was added and incubated for 24 hours. Cell viability was measured, and absorbance was measured at 450 nm with a plate reader using EZ-Tox reagent (Daeil Lab Service, Korea). As a control, a medium in which nothing was treated and a medium in which only glutamate was treated in 10 mM was set. These results are shown in Fig.

HT-22 cells cultured in the same manner as above were treated with 2.8 mM, 8.3 mM and 25.0 mM of lethypoline (DMSO solution), respectively. After 2 hours of treatment, 10 mM of glutamate was added and cultured in an incubator with 5% CO ₂ condition at 37 ° C for 24 hours. The cells were cultured using Operetta (high content imaging system, Perkin Elmer) Images were taken and cell viability was confirmed. As a control, a medium in which nothing was treated and a medium in which only glutamate was treated in 10 mM was set. These results are shown in FIG. 5, and the scale bar in FIG. 5 is 100 μm.

4 to 5, it can be seen that the latofolin according to one aspect of the present invention has an effect of increasing the survival rate of neurons killed by glutamate.

In particular, according to FIG. 5, it can be visually confirmed through the image that most of neuron cells die when glutamate is treated. However, in the case of treatment with lattipone of the present invention simultaneously with glutamate, it was confirmed that the cells were alive at a level similar to that of the control group treated with DMSO only.

In addition, according to the results of FIGS. 4 to 5, the latofolin of the present invention protects nerve cells from oxidative stress and inhibits the death of nerve cells, thereby causing neuronal death or inducing oxidative stress Prevent, or ameliorate a neurological disorder that may be associated with the disease.

According to the usual knowledge of the present invention (see non-patent documents 2, 3 and 4), glutamate corresponds to a major neurotransmitter in the central nervous system and, when present at a high concentration, . Conventional artisans have been used to study the pathogenesis of neurodegenerative diseases by applying this glutamate-induced oxidative stress model to HT22 cells. In accordance with one aspect of the present invention, latifulin or a strong-flavored fragrance extract containing the same exhibits an effect of inhibiting glutamate-induced neuronal cell death in HT22 cells. Therefore, Prevention, or amelioration of these neurodegenerative diseases. ≪ Desc / Clms Page number 2 >

Formulation examples of compositions according to one aspect of the invention are described below, but are also applicable to various other formulations, which are not intended to be limiting but rather to illustrate only specific details.

[Formulation Example 1] Soft capsule

Vitamin E-vitamin C-palm oil-vegetable hardening oil-lecithin is mixed at a ratio of 6: 3: 3: 1: 3: 3 according to a conventional method to prepare a soft capsule filling liquid. 500 mg per capsule is filled to prepare a soft capsule. Separately from this, a soft capsule sheet is prepared in a ratio of 66 parts by weight of gelatin, 24 parts by weight of glycerin and 10 parts by weight of sorbitol solution, and filled with the filling liquid to prepare a soft capsule containing 500 mg of the composition according to the present invention.

[Formulation Example 2] Tablets

20 mg of latifolin, 20 mg of vitamin E, 20 mg of vitamin C, 200 mg of galactooligosaccharide, 60 mg of lactose and 140 mg of maltose are mixed, granulated using a fluidized bed drier, and 6 mg of sugar ester is added. These compositions are tableted by conventional methods to prepare tablets.

[Formulation Example 3] Drinking agent

Lactifolin 50 mg, Vitamin E 20 mg, Vitamin C 20 mg, glucose 10 g, citric acid 0.6 g, and liquid oligosaccharide 25 g are mixed, and 300 ml of purified water is added to each bottle to fill 200 ml each bottle. It is filled in a bottle and sterilized at 130 ° C for 4 to 5 seconds to prepare a drink.

[Formulation Example 4]

50 mg of latifolin, 20 mg of vitamin E, 20 mg of vitamin C, 250 mg of anhydrous crystalline glucose and 550 mg of starch are mixed and granulated into granules using a fluidized bed granulator and filled in a capsule to prepare granules.

[Formulation Example 5]

Injections were prepared in a conventional manner according to the composition shown in Table 1 below.

Compounding ingredient content Latifolin 25 mg Sterile sterilized water for injection Suitable amount pH adjusting agent Suitable amount

[Formulation Example 6] Health functional food

Health functional foods were prepared according to the conventional methods according to the compositions shown in Table 2 below.

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

Although the composition ratio of the above-mentioned vitamin and mineral mixture is relatively well mixed with a composition suitable for health functional foods in a desirable composition, the compounding ratio may be arbitrarily modified.

[Formulation Example 7] Health drinks

Health drinks were prepared according to the compositions shown in Table 3 below in a conventional manner.

Compounding ingredient content Latifolin 10 mg Citric acid 1000 mg oligosaccharide 100 g Taurine 1g Purified water Balance

The above components are mixed according to a conventional health drink manufacturing method, and the mixture is stirred and heated at 85 DEG C for about 1 hour, and the solution thus prepared is then filtered to sterilize.

Claims (14)

A composition for protecting or inhibiting neuronal cell death from glutamate toxicity, the composition comprising latifolin as an active ingredient, wherein the latifolin concentration is 0.01 mu M to 1M concentration based on the total volume of the composition. delete delete A composition for treating, preventing or ameliorating a neurodegenerative disease caused by glutamate toxicity, the composition comprising latifolin as an active ingredient and the latifolin at a concentration of 0.01 mu M to 1M based on the total volume of the composition. 5. The composition of claim 4, wherein the neurodegenerative disease is at least one selected from the group consisting of stroke, amyotrophic lateral sclerosis (Lou Gehrig's disease) and Parkinson's disease. The method according to any one of claims 1, 4, and 5,
Lt; RTI ID = 0.0 > lectifin < / RTI > The method according to claim 6,
Wherein the latifolin is isolated or purified from a strong-flavored ethanol extract. 8. The method of claim 7,
Latiporin is obtained by fractionating the strong-spirited ethanol extract with methylene chloride, fractionating the methylene chloride fraction with hexane-ethyl acetate mixed solvent again, and isolating it with chromatography. The method according to claim 6,
Gangjin is a strong forest 　 pedunculata (L.) Miquel . ) Core of leaf, flower, stem, fruit, root, stem or root of wood, and a composition thereof. 10. The method of claim 9,
Wherein the composition is a core of a stem or root of a hardwood tree. delete delete delete The method according to claim 4 or 5,
Wherein said composition is a pharmaceutical composition.

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