KR101588452B1 - Composition for inflammatory neurodegenerative diseases comprising purple sweet potato terminal shoot extract - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating or treating an inflammatory neurodegenerative disease comprising an extract of Liliaceae as an active ingredient. The present invention also relates to a composition for antiinflammation or antioxidation comprising an extract of Liliaceae .

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or treating an inflammatory neurodegenerative disease,

The present invention relates to a composition for preventing, alleviating or treating an inflammatory neurodegenerative disease comprising an extract of Liliaceae as an active ingredient. The present invention also relates to a composition for antiinflammation or antioxidation comprising an extract of Liliaceae .

This study is the result of the research carried out by the Ministry of Commerce, Industry and Energy and the Korea Industrial Technology Development Agency in the promotion of local specialized industries.

In general, neuroinflammation is accompanied by the onset of various neurological and neurodegenerative diseases such as cerebral ischemia, Alzheimer's disease, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis. Microglia, a unique immune cell in the brain, play a particularly important role in brain inflammation through the production of proinflammatory cytokines, nitric oxide (NO) and other neurotoxic factors. Neuronal neurons can be directly injured by activated microglial cells by stimulating tumor necrosis factor (TNF-α) and cytokines such as interleukin IL-6.

Therefore, inhibition of cell signaling by microglial cells may lead to improvement of neuroinflammatory diseases.

Lipopolysaccharide (LPS) has been widely used as an active model immunostimulant in vivo and in vitro studies. Previous studies have shown that LPS stimulates microglial cells and induces cytokines such as inducible nitric oxide synthase (iNOS), NO and interleukin (IL) -1, IL-6 and tumor necrosis factor (TNF-α) It was proposed to produce.

Korean Patent Laid-Open No. 1020120011981 relates to a dermal extract having anti-inflammatory effect and a composition for external application for skin comprising the same, wherein the dermal extract increases the expression of hemoxigenase-1 and sirtuin-1, It has an inflammatory effect.

The present invention has been made in view of the above needs, and an object of the present invention is to provide an anti-inflammatory or antioxidant composition.

It is another object of the present invention to provide a composition for the treatment or prevention of inflammatory neurodegenerative diseases.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of inflammatory neurodegenerative diseases, comprising an extract of Liliaceae as an active ingredient.

In one embodiment of the present invention, the composition is preferably, but not limited to, inactivating the inducible nitric oxide synthase (iNOS) and NO production.

In another embodiment of the present invention, the extract is preferably an extract of ethyl acetate, but not limited thereto.

In another embodiment of the present invention, the composition preferably inhibits microglial cell activity in microglial cells, but is not limited thereto.

In another embodiment of the present invention, the composition preferably inhibits TNF-a activity, but is not limited thereto.

In addition, the present invention provides a food composition for relieving inflammatory neurodegenerative diseases, comprising an extract of Liliaceae as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for antiinflammatory activity, which comprises the extract of Liliaceae as an active ingredient.

The present invention also provides a functional food composition for antiinflammatory activity, which comprises an extract of Zomotelzici as an active ingredient.

In addition, the present invention provides a composition for antioxidation comprising an extract of Liliaceae as an active ingredient.

The pharmaceutical composition comprising the extract of Liliaceae according to the present invention as an active ingredient may be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned effective ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, A binder, a coating agent, a swelling agent, a lubricant, a lubricant or a flavoring agent.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, natural sugars such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

In addition, the present invention provides a food composition comprising the above extract.

The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meats, chocolates, foods, confectionery, pizza, ramen noodles, gums, ice cream, alcoholic beverages, vitamin complexes, .

The present invention provides a health functional food comprising the composition.

The present invention also provides the use of a composition comprising the above-described extract of Liliaceae for the preparation of a medicament or food for the prevention and treatment of tuberculosis as an active ingredient. The composition of the present invention comprising the extract of Jamiji end as an active ingredient can be used for the preparation of medicines or foods for the prevention or treatment of diseases related to prevention and treatment of tuberculosis.

The present invention also provides a method of preventing or treating a tuberculosis disease comprising administering to a mammal a therapeutically effective amount of a Jamie endum extract.

The term "mammal " as used herein refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.

As used herein, the term "therapeutically effective amount " refers to the amount of active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, This includes an amount that induces relief of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like.

The dose of the Jamaicans extract, which is an active ingredient in the pharmaceutical composition of the present invention, may be 6 to 30 mg per kg of body weight once or three times daily, depending on the patient's age, sex, symptom, administration method or preventive purpose . Dosage levels for patients exhibiting the specific symptoms 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.

In the treatment method of the present invention, the composition comprising the extract of Liliaceae of the present invention as an active ingredient may be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, And can be administered in a conventional manner.

INDUSTRIAL APPLICABILITY The pharmaceutical composition comprising the extract of Jamiji, which is an industrial by-product according to the present invention, can weaken LPS-induced inducible NO synthase and NO production in BV-2 cells, and inducible nitrogen (INOS) protein expression. These results suggest that the extract of Zygomycetes can be used as an effective anti-inflammatory, antioxidant and neurodegenerative disease.

FIG. 1 is a diagram illustrating a process of extracting the end of a line.
Figure 2 compares the antioxidant activities of the extract fractions from the end of the jam.
FIG. 3 is a graph showing the inhibition of NO production (BV-2 cells) of the endermic lot extract.
FIG. 4 is a graph showing cell viability (BV-2 cells) of the endermic lot extract. FIG.
FIG. 5 is a graph showing the antioxidative effect of the extract of Lycopersicon esculentum.
FIG. 6 is a graph showing the efficacy against inflammatory proteins caused by the extract of L. japonica.
FIG. 7 is a graph showing inhibition of TNF-alpha production (BV-2 cells) by the end-tails extract.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Example  One: Jami end  extraction

To increase the availability of sweet potatoes, it is possible to use sweet potatoes at the end (sweet potato trunks or leaves attached to the trunks about 10 to 15 cm from the growth point of the branches). The end of the sweet potato is 10 ~ 15 cm long at the end of the stem including the leaf and petiole in the bud which grew after 50 ~ 60 days after straightening the trout.

Jami is a sweet potato cultivar determined in 1998 as a cultivated or pigmentable cultivar after it has been selected and cultivated by artificial hybridization of Yamasawa Murasaki with anthocyanin pigment in 1993 as a model.

The end of Jami was purchased from Jecheon farmhouse in Chungcheongbuk-do, and 1 kg of distilled water was added to 100 g of Jamie end, followed by hot water extraction at 80 캜 for 12 hours. 2 The aqueous solution filtered through filter paper was extracted with 5 L of ethanol.

The extracted ethanol extracts were firstly adsorbed on polyphenols using HP20 adsorbent, washed, and then eluted with ethanol. In order to separate the components of the end-tails extract, they were sequentially fractionated using various solvents (Fig. 1). After 100 ml of n-hexane, 100 ml of n-hexane was added to the extract, and the mixture was stirred for 30 minutes. Then, the mixture was subjected to n-hexane fractionation, followed by Chloroform fraction-> Ethyl acetate fraction-> Butanol fraction-> - > DMSO.

    As shown in FIG. 2, the polyphenol content in chloroform and ethyl acetate fraction was high. Especially, the content of polyphenol in ethyl acetate was significantly higher than that of other solvent fractions Respectively.

Example  2: Antioxidant

Many of the functions of the living body are based on the antioxidant mechanism, and polyphenols can act as antioxidants, and the antioxidant activities of the endophytic fractions with different polyphenol contents were measured. Antioxidant activity was measured by electron donating ability. The antioxidant activity by electron donating activity was evaluated by using free radicals such as DPPH, and directly assessing the reactivity of the antioxidants against radicals in the model system of LOO from the decrease or decrease rate of radicals. Or DPPH forms an irreversibly stable molecule by electrons or hydrogen from the antioxidant, so that the antioxidant activity can be measured from the electron donating ability. In this experiment, electron donating ability (EDA) was measured by Blois et al. Namely, a ⁴ × 10 ^-4 M DPPH solution (1.1-diphenyl-2-picrylhydrazyl) was added to 0.2 ml (6.0 mg / ml) of each sample and the absorbance was measured at 525 nm using a spectrophotometer after 10 minutes.

The DPPH radical scavenging activity of the extract fraction, Jamae extract, showed that the antioxidative activities of chloroform and ethyl acetate fraction were similar to those of the other fractions. The ethyl acetate fraction showed the highest activity (Fig. 2)

Example  3: cell survival rate

The effect of LPS and extracts of B. subtilis ethyl acetate in BV-2 cells stimulated with LPS on cell viability was confirmed. As a result of cell viability measurement, it was confirmed that the cell survival rate was not changed in all experimental groups treated with LPS and Jamaicana extract alone or in the same manner (FIG. 4).

This indicates that LPS, an inflammation inducing substance, and Jamaicans extract, a liver function enhancing substance, do not affect cell survival.

Example  4: LPS In activated glial cells Extract  Concentration-dependent NO  Production-inhibiting action

In order to analyze the anti - inflammatory effect of the extract of L. japonica L., we investigated whether LPS, an inflammation - inducing factor, is dependent on the concentration of NO produced in stimulated glial cells.

   NO was measured by Griess reagent and NO was induced by LPS (5 ug / ml) in neurons. The NO was about 34 mM, which was about 30 times higher than that of 1 mM used as a control. As a result of treatment with the extracts at the end, the NO production was reduced to 100, 200, 400, and 800 ug / ml, respectively, depending on the concentration of the endermic extract.

Example  5: in the glial cells Jami end  After the extract treatment LPS Induced by iNOS  inducible nitric oxide synthase ) Concentration-dependent inhibition of protein expression

Changes in iNOS protein expression patterns were observed after administration of the extracts from the end of the. The expression of iNOS protein was analyzed by LPS, and the iNOS protein was expressed by activating the glial cells. Immunoblot analysis showed that the expression level of the iNOS protein in the control group not treated with LPS was negligible (Fig. 6, right panel, lane 1)

The expression of iNOS protein was significantly increased in the LPS-treated experimental group (Fig. 6, right panel, lane 2), indicating that the concentration-dependent decrease in expression was observed after treatment with the endermic extract (Fig. , 100 ug / ml Jamie end extract, lane 4, 200 ug / ml Jamie end extract). Beta-actin was used as a control for protein quantitation (Fig. 6).

Example  6: TNF - Alpha cytokines

TNF-alpha, a representative inflammatory cytokine, was measured using a kit in order to determine whether the ethyl acetate material inhibits inflammatory cytokines. This confirms the effect of ethyl acetate on the IL-6 concentration. As shown in FIG. 7, TNF-alpha, which is an inflammatory cytokine, was measured after LPS treatment, and TNF-alpha was significantly increased in the LPS-treated group, but it was decreased in the group treated with ethyl acetate. Therefore, it was confirmed that the end-tails ethyl acetate material decreased the inflammatory cytokine TNF-alpha in a dose-dependent manner.

Claims (11)

A method for reducing inducible nitric oxide synthase protein expression in microglial cells in vitro by treating lipopolysaccharide (LPS) -treated microglia in vitro. delete      The method according to claim 1, wherein the extract is a Zomi-endone ethyl acetate extract.   A method for reducing TNF-alpha expression of microglial cells in vitro by treating Jasmine Leaf Extract with in liposomal lipopolysaccharide (LPS) -treated microglial cells. 5. The method according to claim 4, wherein the extract is a Zygomycetes ethyl acetate extract.

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