KR20190025289A - A composition for antiinflammatory and inflammatory neurodegenerative diseases comprising jujube seed extract - Google Patents

Abstract

The present invention relates to an anti-inflammatory composition for preventing or treating an inflammatory neurodegenerative disease, including a jujube seed extract, which can attenuate production of inducible nitrogen oxide synthase (iNOS) and NO. The composition of the present invention comprises a jujube seed extract as an active ingredient, and inhibits microglia activity.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or treating inflammation and inflammatory neurodegenerative diseases, which comprises a jujube extract, a composition for preventing or treating inflammation and inflammatory neurodegenerative diseases,

The present invention relates to a composition for preventing or treating anti-inflammatory and inflammatory neurodegenerative diseases, comprising a jujube extract.

Jujube ( Zizyphus) jujuba Miller, jujube) is a fruit of deciduous and broad-leaved trees of the genus Zadigus and Zizyphus, distributed in Korea, Europe and Asia. It is known that there are about 40 varieties and 400 varieties in tropical, subtropical and temperate regions. Jujube has long been known to be effective in the treatment of calming stabilizers, alleviating digestion, celiac treatment, and tonic action.

Jujube seed is a byproduct of jujube processing, and has been used as a material for medicinal and processed foods. The content of carbohydrate was 89.4%, and the content of minerals such as magnesium, manganese, and potassium and amino acids such as proline, aspartic acid, and unsaturated fatty acids such as oleic acid and linoleic acid were high . The medicinal components of jujube seeds are known to be betulin and betulinic acid. Especially, cyclic-adenosine monophosphate (c-AMP) is known to be about 1,000 times higher than other plants.

On the other hand, there are antioxidant enzymes that protect against reactive oxygen species in the body such as superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT), glutathione reductase and glutathione-S- transferase. However, The role of antioxidants in the human body due to substances, smoking, stress, etc., exceeds the defense system, leading to protein degradation and DNA damage

       Therefore, synthetic materials such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) have been developed as antioxidants. However, studies on finding dietary antioxidants that are safe from nature have been actively carried out Become

       Inflammation, on the other hand, is a defense mechanism to prevent damage to human tissues caused by infection, which involves symptoms such as development, fever and pain. If the inflammatory reaction is repeated for a long period of time, it can cause various chronic diseases such as neurodegenerative diseases and cancer.

       Macrophages are involved in and regulate the production of inflammatory mediators such as nitric oxide (NO), prostaglandin (PG) and pro-inflammatory cytokines in the human immune response. These inflammatory mediators induce an inflammatory response and, if the host immune response does not respond appropriately, induce inflammatory diseases.

Korean Patent Laid-Open Publication No. 1020120011981 discloses 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 hemeoxygenase-1 and sirtuin-1 It has an anti-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 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 achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating inflammatory neurodegenerative diseases, comprising an extract of Daechu seed as an active ingredient.

In one embodiment of the invention, the composition is preferably but not limited to weaken inductive nitrogen oxide synthesis (iNOS) and NO production.

In another embodiment of the present invention, the extract is preferably a jujube alcohol extract, more preferably an ethanol extract, but is 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 addition, the present invention provides a food composition for relieving inflammatory neurodegenerative diseases, comprising an extract of Daechu seed as an active ingredient.

The present invention also provides a pharmaceutical composition for antiinflammation comprising an extract of Daechu Seed Extract as an active ingredient.

The present invention also provides a functional food composition for antiinflammation comprising an extract of Daechu seed as an active ingredient.

The present invention also provides a composition for antioxidation comprising an extract of Daechu seed as an active ingredient.

The pharmaceutical composition comprising the jujube extract of the present invention as an active ingredient may be prepared 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 sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium 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.

The present invention also provides a food composition comprising the jujube 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 jujuba extract as an active ingredient for the manufacture of a medicament or food for the prevention and treatment of tuberculosis. The composition of the present invention comprising the jujube extract as an active ingredient can be used for the manufacture of medicines or foods for the prevention or treatment of diseases associated with tuberculosis prevention and treatment.

The present invention also provides a method of preventing or treating inflammatory diseases, comprising administering to a mammal a therapeutically effective amount of a jujube 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 jujube extract which is an active ingredient in the pharmaceutical composition of the present invention can be 6 to 30 per kg of body weight once or three times a day depending on the patient's age, sex, symptom, method of administration, or prophylactic purpose have. 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 jujube extract of the present invention as an active ingredient may be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intramuscularly, transdermally, topically, And can be administered in a conventional manner.

The pharmaceutical composition comprising the extract of Daechujang which is an industrial by-product according to the present invention can completely weaken the production of LPS-induced inducible nitric oxide synthase (iNOS) NO in BV-2 cells and these results show that the jujube extract is effective Anti-inflammatory agents and neurodegenerative diseases.

Figure 1 shows the DPPH inhibition of jujube seed extract.
Figure 2 shows the cell viability (BV-2 cells) of the jujube extract.
FIG. 3 is a graph showing inhibition of NO production by the jujube seed extract. FIG.

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 1: Jujube Seed Extraction

Daechu was purchased from Taewoong Foods Co., Ltd., and 100 g of jujube seeds were extracted with 1 L of ethanol for 72 hours at room temperature.

Example 2: Polyphenol content of jujube seed extract

The content of polyphenols in the jujube extract was determined by Folin-Denis method.

Dissolve 1 mg of the jujube extract of Example 1 in 1 mL of distilled water, add 2 mL of a 2-fold diluted Folin reagent (manufactured by Sigma Aldrich) to 2 mL of the diluted solution 10 times with distilled water, mix well, . Thereafter, ₂ mL of 10% Na ₂ CO _{3 was} added and reacted for 1 hour. Then, the absorbance was measured at 700 nm using a UV-Vis spectrophotometer (UVIKON 922, Kontron, Italy) Respectively.

At this time, a standard curve using gallic acid was prepared by measuring the absorbance at 700 nm in the same manner as above so that the final concentration of gallic acid was 0, 5, 25, 50 or 75 μg / mL.

The content of polyphenol present in the jujube extract was measured using gallic acid as a reference material. The results of repeated experiments were shown in Table 1 below. The results are shown in Table 1, and the standard deviation (mean ± SD) Respectively.

The total polyphenol content of the jujube extract was about 26.12 mg / g.

Sample Total polyphenol content
(μg / mg) Jujube Seed Extract 26.12 ± 1.12

Example 3: Antioxidant

The DPPH radical scavenging activity of the jujube extract was determined by adding 30 μL of the extract solution and 30 μL of the 60 μM DPPH solution dissolved in ethanol to the capillary tube for 2 min at room temperature. The electron spin resonance (ESR) spectrometer -PX 2300, JEOL, Japan). The ESR spectrophotometer measured 336.5 ± 5 mT for the magnetic field, 5 mW for the microwave power, 9.41 GHz for the modulation frequency, DPPH free radical scavenging activity (%) for the modulation antioxidant sample (ESR signal intensity for As a result of calculation with X100,

Showed a DPPH radical scavenging activity of more than 63% at a concentration of 250 μg / mL and a DPPH radical scavenging activity of more than 90% at a concentration of 500 μg / mL (FIG. 1).

Example 4: Cell viability

In order to examine the effect of LPS and jujube extract on cell survival in BV-2 cells stimulated with LPS, cell biability was measured by using 3- [4,5-Dimethylthiazol-2-yl] -2,5-diphenyl-tetrazolium bromide MTT) assay.

Cells (1 × 10 ⁵ cells / ml) were seeded in a 96-well plate (180 μL), cultured in a CO2 incubator for more than 12 hours, and cultured for 24 hours. After culturing, the culture medium was removed, and 200 μL of the medium containing 0.5 mg / mL MTT was added, followed by incubation for 4 hours to reduce the MTT. After removing the culture medium, 100 μL of dimethylsulfoxide (DMSO) was added and the resulting formazone crystals were dissolved. The absorbance was measured at 540 nm using an ELISA reader. Cell viability was expressed as percentage (%) as compared with control.

As a result of cell viability measurement, it was confirmed that cell survival rate was not changed in all experimental groups treated with LPS and jujube extract alone or in the same way (FIG. 2).

This indicates that the inflammation inducers LPS and jujube extract do not affect cell survival.

Example 5: Concentration-dependent inhibition of NO production of jujube extract in LPS-activated glial cells

In order to investigate the anti - inflammatory effect of the extract of Daechu Seed Extract, LPS, which is an inflammation inducer, was examined to determine whether or not the concentration of nitric oxide synthase (NO)

NO was determined by seeding 5 × 10 ⁵ cells / well in a 24-well plate, adding LPS and extracts at different concentrations, reacting in a 24 h incubator, and then adding 50 μL each of Griess reagent (1% sulfanilamide / After reacting with 0.1% N- (1-naphthyl) -ethylenediamine dihydrochloride / 2.5% H3PO4), the value is measured using a Sunrise basic 96-well microplate spectrophotometer (TECAN AUSTRIA) with a wavelength of 540 nm.

The number of NO induced by LPS (100 ng / mL) in the glial cells was about 23 μM, which was about 23 times higher than that of 1 μM used as a control. , 250, and 500 μg / mL, respectively, indicating that NO production was reduced in a concentration-dependent manner in the jujube extract (Fig. 3).

Claims (5)

A composition for treating or alleviating an inflammatory neurodegenerative disease comprising an extract of Daechu seed as an active ingredient. The composition for treating or alleviating inflammatory neurodegenerative diseases according to claim 1, wherein the composition weakens inducible nitric oxide synthase (iNOS) and NO production. The composition for treating or alleviating an inflammatory neurodegenerative disease according to claim 1, wherein the extract is a jujube alcohol extract. The composition for treating or alleviating inflammatory neurodegenerative diseases according to claim 1, wherein the composition inhibits microglial cell activity in microglial cells. A composition for antiinflammation comprising an extract of Daechu seed as an active ingredient.

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