KR101385381B1 - Composition for preventing and treating inflammatory diseases comprising water derived from jeju - Google Patents

Abstract

The present invention relates to a composition for preventing and treating inflammatory diseases containing Jeju water as an active ingredient, wherein the Jeju water is cyclooxygenase-2 (COX-2) dependent on inflammatory and allergic reactions. a prostaglandin _{D 2 (prostaglandin D 2, PGD} 2) inhibiting the generation and 5-lipoxygenase (5-lipoxygenase; 5-LOX ) dependent leukotriene C ₄ (LTC ₄₎ By inhibiting production and at the same time inhibiting the degranulation response from mast cells, it can be usefully used for the prevention and treatment of inflammatory or allergic diseases.

Description

Composition for preventing and treating inflammatory diseases comprising water derived from jeju}

The present invention relates to a composition for preventing or treating inflammatory diseases comprising Jeju water as an active ingredient effective for the prevention or treatment of inflammatory diseases including allergic diseases.

Asthma is a chronic inflammatory disease caused by representative inflammatory cells such as mast cells and various humoral inflammatory mediators such as eosinophils and T-lymphocytes. It is characterized by reversible airway obstruction and bronchial hypersensitivity. Get involved.

Substances involved in asthma are inflammatory, other than leukotriene (LT), C ₄ , D ₄ , and E ₄ , a metabolite of 5-lipoxygenase, known as the slow reacting substance of anaphylaxis (SRS-A). It is known that cytokines such as Interleukin (IL) -4, -5, -6 are involved (Barnes P. J, et al., Inflammatory mediators of asthma: an update, Pharmacol Rev, 50, pp 515-596, 1998; Galli S, et al., Mast-cell-leukocyte cytokine cascades in allergic inflammation, Allergy 50, pp 851-862, 1995).

Bronchial asthma is also a disease of inflammatory reactions. Inflammation is a defense of biological tissues against local injuries. That is, in response to various harmful stressors, the biological defense reaction to recover the original state by removing the injury caused by stimulation is an inflammatory reaction. Inflammatory stimuli include infection, chemical and physical stimulation. Biocomponents related to the inflammatory reaction include low-molecular or polymeric chemicals such as free radicals, proteins, saccharides and lipids, and plasma, blood cells, blood vessels, And connective tissue.

The process of inflammation can be divided into acute inflammation and chronic inflammation. Acute inflammation is a short-term reaction within a few days. Plasma components and blood cells are involved in dephosphorylation by displaying a microcirculatory system. Chronic inflammation has a long duration and proliferation of tissues (Coico R, et al., Immunolgy-a Short Course, Wiely-Liss, INC. Fifth Edition, p16-18, 2003).

Key parameters for inducing the inflammatory and allergic diseases material prostaglandin acids (prostaglandines), leukotriene acids (leukotriens), platelet activating factor (PAF) Phospholipase A ₂ (phospholipase A _2), such as, cyclooxygenase (cyclooxygenase) And arachidonic acid, which is a precursor, by lipoxygenase.

Prostaglandins bind to specific cell surface receptors and act to increase intracellular concentrations of cAMP (and possibly cGMP). The effect of increasing cAMP depends on cell type and PGA ₂ , PGB _{2 lower} blood pressure (Cicardo VH, Mechanism of the hypotensive effect of prostaglandins A2 and E2, Acta. Physiol. Lat. Am, 1981; 31, pp 85-91, 1981; Liu F, et al., Prostaglandin B2-induced pulmonary hypertension is mediated by TxA2 / PGH2 receptor stimulation.Am. J. Physiol, 267, pp 602-608, 1994), PGD ₂ , PGE ₁ (Gollman HM et al., Comparative pyrogenic potency of endogenous prostanoids and of prostanoid-mimetics injected into the anterior hypothalamic / preoptic region of the cat.Brain Res. 449, pp281-293 , 1988, Collins DR, et al., Cutaneous sympathetic motor rhythms during a fever-like response induced by prostaglandin E (1), Neuroscience, 110, pp 351-360, 2002).

In particular, PGD ₂ is known to exacerbate asthma by contracting smooth muscle in bronchial asthma patients (Shiraishi Y, et al., Cyclooxygenase-2 / prostaglandin D2 / CRTH2 pathway mediates double-stranded RNA-induced enhancement of allergic airway inflammation, J. Immunol., 180, pp 541-549, 2008). Leukotrienes (LT) constitute the local functional hormone group produced in vivo from arachidonic acid. Important leukotrienes include leukotriene B ₄ (LTB ₄ ), leukotriene C ₄ (LTC ₄ ), leukotriene D _{4 4} ) and leukotriene E ₄ (LTE ₄ ). The biosynthesis of these leukotrienes is initiated by the enzyme 5-lipoxygenase acting on arachidonic acid to produce an epoxide known as leukotriene A ₄ , which can be converted to other leukotrienes (LTB ₄ , LTC ₄ , LTD ₄ , LTE ₄ ). Leukotriene is involved in allergic and allergic reactions such as pulmonary artery disease such as asthma, chronic bronchitis and related obstructive airways disease, allergic rhinitis, contact dermatitis, allergic conjunctivitis, inflammation such as arthritis or inflammatory bowel disease, pain .

Recently, drugs that have attracted attention as an allergic asthma treatment are drugs that have both histamine free inhibition, leukotriene C ₄ production inhibition, and platelet activator inhibitory activity (Dahlen SE, Treatment of asthma with antileukotrienes: first line or last resort therapy). , Eur. J. Pharmacol., 533, pp 40-56, 2006; Kitamura S, Leukotriene (B4, C4, D4, E4), Nippon Rinsho., Suppl 8, pp28-33, 2005).

On the other hand, natural natural water has a side effect even if taken for a long time and its efficacy has been proven for a long time, there is an advantage that the selection of raw materials is very easy. Therefore, the present invention was intended to complete the present invention using natural natural water as a solution for reducing the above side effects and at the same time to obtain a pharmacological effect.

Accordingly, the present inventors have completed the present invention by confirming that Jeju water taken from a specific region of Jeju Island can improve inflammatory diseases including allergy or asthma, as a result of earnest research to overcome the problems of the prior arts. It became.

Accordingly, it is an object of the present invention to provide a composition containing Jeju water as an active ingredient having an inflammatory disease improvement effect including allergy or asthma.

In addition, another object of the present invention to provide a functional food and pharmaceutical composition using the composition for preventing or treating inflammatory diseases, including allergy or asthma, including the Jeju water.

In order to achieve the above object, the present invention provides a composition for preventing or treating inflammatory diseases containing Jeju water as an active ingredient.

The Jeju water is groundwater taken from Jeju Samdasoo or Jeju Island, and is especially ground water taken by digging 200 to 600m underground in any one or two or more areas selected from Jocheon-eup, Gyorae-ri, Seogwipo-si, Jungmun-dong, and Seogwipo-si. According to one embodiment, the groundwater taken by digging at 250m is named S-3, and the groundwater taken by digging at 550m is named S-2.

According to an embodiment of the present invention, by digging 250m underground in Dosun-dong, Seogwipo-si, Jeju-do, it was possible to obtain groundwater taken from 10m underground (average sea level).

Jeju water of the present invention is specifically a mineral containing sodium (Na), magnesium (Mg), calcium (Ca), potassium (K), silica (SiO 2) and the like; Anions including fluorine ion, chlorine ion, nitrogen nitrate, sulfate ion, bromine ion and the like; And vanadium (V), aluminum (Al), arsenic (As), iron (Fe), copper (Cu), zinc (Zn), manganese (Mn), lead (Pb), chromium (Cr), selenium (Se) , Cadmium (Cd), boron (B), molybdenum (Mo), strontium (Sr), rubidium (Rb), nickel (Ni), barium (Ba), thallium (Tl), beryllium (Be), antimony (Sb) And trace elements including uranium (U), titanium (Ti), cobalt (Co), germanium (Ge), tin (Sn), zirconium (Zr), tungsten (W), cesium (Cs), and the like. .

In particular, the vanadium (V) is contained in the content of 7 to 30 ppb (μg / L), preferably in the case of three waters 7.1 to 8.9 ppb (μg / L), for S-2 22.0 to 26.0 ppb (μg / L) In the case of L) and S-3, the content is 24.0 to 28.0 ppb (μg / L), and the strontium (Sr) is included in the content of 10 to 50 ppb (μg / L). To 19.0 ppb (μg / L), 2-2 to 30.0 ppb (μg / L) for S-2 and 35.0 to 45.0 ppb (μg / L) for S-3.

Water is the Cheju inflammation and cyclooxygenase -2 (cyclooxygenase-2; COX- 2) that are related to an allergic reaction inhibition dependent prostaglandin _{D 2 (Prostaglandin D 2, PGD} 2) generation, and 5-lipoxygenase (5-lipoxygenase; 5-LOX) dependent leukotriene C ₄ (LTC ₄ ) By inhibiting production and at the same time inhibiting the degranulation response from mast cells, it can be usefully used for the prevention and treatment of inflammatory or allergic diseases.

The inflammatory diseases include allergic diseases, chronic inflammatory diseases, acute inflammatory diseases, and the like, and more particularly to allergic diseases, inflammatory bowel diseases, systemic lupus erythematosus, inflammatory collagen vascular diseases, glomerulonephritis, inflammatory skin diseases, , Retinitis, gastritis, hepatitis, enteritis, pancreatitis and nephritis.

In addition, the allergic disease may be any one selected from the group consisting of hypersensitivity, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis and urticaria.

In the composition for preventing or treating an inflammatory disease of the present invention, the composition is Jeju water itself as a functional food having an effect of improving inflammatory diseases including allergy or asthma, as it has anti-inflammatory, anti-asthmatic and anti-allergic effects. It is also available in addition to other functional foods containing natural extracts that have an effect on improving inflammatory diseases or in the form of extracting these natural products.

The functional food of the present invention can be used as a health functional food or health beverage, it is preferable to use in the form of soft or hard capsules filled with functional powders, pills, tablets or dry powdered ingredients.

Examples of foods to which Jeju water of the present invention can be added include various foods, candy, chocolate, beverages, gums, teas, vitamin complexes, health supplements, and the like, and are powders, granules, tablets, pills, capsules or beverages. Available in form.

The health beverage of the present invention has no particular limitation on the liquid component except that it contains Jeju water as an essential ingredient in the ratio indicated, and may contain various flavors or natural carbohydrates, etc. as additional ingredients, like ordinary drinks. Examples of natural carbohydrates include monosaccharides such as disaccharides such as glucose and fructose, and polysaccharides such as maltose and sucrose. For example, conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of the flavors include natural flavors such as tau martin, stevia extract, heba dioside A, glycyrrhizin and synthetic flavors such as saccharin and aspartame.

In addition to the above, the composition of the present invention may also be used as a flavoring agent, coloring agent, enhancer, pectic acid and its salt, alginic acid and its salt, organic acid, protective colloid thickening agent, pH of various nutrients, vitamins, minerals, synthetic flavors and natural flavors, Stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.

Jeju water of the present invention is a natural natural water, there is almost no toxicity and side effects can be used with confidence even for long-term use for the purpose of prevention.

When the Jeju water of the present invention is added to food for the purpose of improving inflammatory disease, it may generally be added at 20 to 70% by weight of the total food weight, and when added to a beverage, 1 to 100 based on 100 mL mL, preferably in a content ratio of 10 to 80 mL.

The present invention also provides a pharmaceutical composition for preventing or treating inflammatory diseases, which contains the Jeju water of the present invention as an active ingredient.

The pharmaceutical composition according to the present invention preferably contains 1 to 70 parts by weight of Jeju Water based on the total weight of the pharmaceutical composition. If Jeju water is contained in a small amount out of the above content range, it is difficult to obtain a desired inflammatory disease treatment or prophylactic effect, whereas if it is contained in an excessive amount, treatment or prophylactic effect of inflammatory disease may be slowed down.

In addition, the pharmaceutical composition according to the present invention may further comprise an appropriate carrier, excipient or diluent conventionally used in the production of the pharmaceutical composition.

Examples of the carrier, excipient or diluent which can be used in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method .

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 formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose sucrose), lactose, gelatin, and the like.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The amount of Jeju water used in the present invention may vary depending on the route of administration, the degree of disease, the age, sex, and weight of the patient, and may be administered once to several times daily.

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes. All modes of administration may be expected, for example, by oral, nasal, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.

Jeju water of the present invention, inflammation and cyclooxygenase -2 (cyclooxygenase-2; COX- 2) that are related to an allergic reaction inhibition dependent prostaglandin _{D 2 (Prostaglandin D 2, PGD} 2) generation, and 5-ripok Shigenase (5-lipoxygenase; 5-LOX) dependent leukotriene C ₄ (LTC ₄ ) Since it inhibits the production and at the same time inhibits the degranulation reaction from mast cells, it can be usefully used for the prevention or treatment of inflammatory diseases including asthma or allergy.

Figures 1a to 1c is to examine the effect of inhibiting degranulation reaction from mast cells of Samdasoo, S2 and S3 which is Jeju water according to the present invention,
2a to 2c is a review of the effects of inhibiting the production of 5-LOX-dependent LTC ₄ of Samdasoo, S2 and S3, which are Jeju water according to the present invention,
3A to 3C illustrate the effects of inhibiting COX-2 dependent PGD ₂ production of Samdasoo, S2 and S3, which are Jeju water according to the present invention.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are illustrative of the present invention, and the contents of the present invention are not limited by these examples.

Example 1 Preparation of Jeju Water

1. Preparation of Samdasoo

Location of Water: Gyorae-ri, Jeju-si

Date of manufacture: 2010.02.28

The ground water collected at 420m above sea level and Gyorae-ri, Jocheon-eup, Jeju-si, Jeju-do, was removed from the microfiltration system using a microfiltration of less than 1µm. filtration) Low molecular weight, high molecular and colloidal materials were filtered using a porosity filter of tens to hundreds of nanometers (nm) using 0.45 ㎛ and 0.2 ㎛. The ground water thus treated was passed through a sterilized ultraviolet water sterilizer that emits strong ultraviolet rays having a wavelength of 253.7 nm to obtain final ground water (number of products).

2. S-2 Preparation

Location of Water: Jungmun-dong, Seogwipo-si

Date of manufacture: 2009.10.14

Groundwater collected from Jungsan-dong, Jungmun-dong, Seogwipo-si, Jeju-do, Korea, was transported by tank lorry and transported to groundwater using a tank lorry, and then used as a microporous filter of 0.45㎛ and 0.2㎛ for a pore filter of tens to hundreds of nanometers (nm). Low molecular weight materials and polymer and colloidal materials were filtered out. The ground water thus treated was passed through a sterilization process through a UV-derivative sterilizer that emits strong ultraviolet rays having a wavelength of 253.7 nm, and finally filtered through a sterilization filter (0.2 μm) to obtain final ground water (number of products).

3. S-3 Preparation

Location of Water: Dosun-dong, Seogwipo-si

Date of manufacture: 2009.10.14

Groundwater collected at 240m above sea level and excavation depth 250m located in Dosun-dong, Seogwipo-si, Jeju-do was transported using tank lorry and then used a microfiltration filtration 0.45µm and 0.2µm. The molecular weight and the polymer and colloidal materials were filtered out. The ground water thus treated was passed through a sterilization process through a UV-derivative sterilizer that emits strong ultraviolet rays having a wavelength of 253.7 nm, and finally filtered through a sterilization filter (0.2 μm) to obtain final ground water (number of products).

Hereinafter, in the following examples, the experiment was performed using Samdasoo, S-2 and S-3 prepared by the above process.

Example 2 Jeju Water Analysis

1. Physicochemical Analysis

pH and EC (electric conductivity) were analyzed using Orion 5 Star instrument, bicarbonate was analyzed by 0.01N-HCl titration, free carbonate by 0.02N-NaOH titration, and hardness by 0.01M-EDTA titration. The results are shown in Table 1 below.

unit Water quality standard Samdasoo S-2 S-3 pH 5.8-8.5 7.8 ± 0.1 8.3 ± 0.1 8.3 ± 0.1 Hardness mg / L 300 18.0 ± 2.0 27.0 ± 2.5 22.0 ± 2.0 Turbidity NTU One 0.084 ± 0.010 0.087 ± 0.010 0.066 ± 0.010 smell Odorless Odorless Odorless Odorless flavor insipidity insipidity insipidity insipidity Electrical conductivity uS / cm - 70.0 ± 3.0 148.0 ± 5.0 175.0 ± 5.0 Bicarbonate mg / L - 28.0 ± 2.0 78.0 ± 3.0 89.0 ± 3.0 Free carbonic acid mg / L - 2.5 ± 0.5 2.1 ± 0.7 0.5 ± 0.6

2. Anion Analysis

Anion analysis was performed using a DIONEX ICS-2000 instrument, the results are shown in Table 2 below.

unit Water quality standard Samdasoo S-2 S-3 Fluorine (F ^-) mg / L 2 ≪ 0.1 ≪ 0.1 ≪ 0.1 Chlorine (Cl ^-) mg / L 250 6.0 ± 0.5 6.0 ± 0.5 7.0 ± 0.5 Nitric Acid Nitrogen (NO ₃ -N) mg / L 10 0.3 ± 0.05 0.3 ± 0.05 0.1 ± 0.05 Sulfate ion (SO ₄ ^2- ) mg / L 200 1.5 ± 0.3 2.3 ± 0.3 2.7 ± 0.3 Bromate Μg / L - ≪ 1.0 ≪ 1.0 ≪ 1.0

3. Major Mineral Analysis

Major mineral analysis was performed using a Varian 720-ES instrument, the results are shown in Table 3 below.

unit Water quality standard Samdasoo S-2 S-3 Sodium (Na) mg / L - 6.0 ± 1.0 18.0 ± 1.0 5.0 ± 1.0 Magnesium (Mg) mg / L - 2.5 ± 0.7 5.0 ± 1.0 3.0 ± 1.0 Potassium (K) mg / L - 2.0 ± 0.5 5.0 ± 0.5 3.0 ± 0.5 Calcium (Ca) mg / L - 2.6 ± 0.5 2.6 ± 0.5 3.4 ± 0.5 Silica (SiO ₂ ) mg / L - 24.0 ± 2.0 22.0 ± 2.0 25.0 ± 2.0

4. Trace element analysis

Trace element analysis was performed using a Varian 830-MS instrument, the results are shown in Table 4 below.

unit Water quality standard Samdasoo S-2 S-3 Vanadium (V) Μg / L - 8.0 ± 0.9 24.0 ± 2.0 26.0 ± 2.0 Aluminum (Al) Μg / L 200 1.5 ± 0.8 10 ± 1.0 10 ± 1.0 Arsenic (As) Μg / L 50 0.5 ± 0.3 3.0 ± 0.7 3.0 ± 0.7 Iron (Fe) Μg / L 300 <0.2 4.0 ± 1.0 1.2 ± 0.5 Copper (Cu) Μg / L 1,000 <0.2 0.4 ± 0.3 0.2 ± 0.3 Zinc (Zn) Μg / L 1,000 0.4 ± 0.3 8.0 ± 0.5 1.7 ± 0.5 Manganese (Mn) Μg / L 300 <0.2 0.2 ± 0.2 <0.2 Lead (Pb) Μg / L 50 <0.2 0.6 ± 0.3 <0.2 Chromium (Cr) Μg / L 50 0.3 ± 0.4 1.0 ± 0.3 0.6 ± 0.3 Selenium (Se) Μg / L 10 <0.2 0.4 ± 0.3 <0.2 Cadmium (Cd) Μg / L 5 <0.2 <0.2 <0.2 Boron (B) Μg / L - 7.1 ± 1.2 15.2 ± 1.5 23.9 ± 1.5 Molybdenum (Mo) Μg / L - 1.0 ± 0.5 3.1 ± 0.7 6.0 ± 0.7 Strontium (Sr) Μg / L - 15.0 ± 4.0 25.0 ± 5.0 40.0 ± 5.0 Rubidium (Rb) Μg / L - 10.0 ± 1.2 8.0 ± 1.2 12.7 ± 1.5 Nickel (Ni) Μg / L - <0.2 <0.2 <0.2 Barium (Ba) Μg / L - 0.6 ± 0.3 1.1 ± 0.5 1.9 ± 0.5 Thallium (Tl) Μg / L - <0.2 <0.2 <0.2 Beryllium (Be) Μg / L - <0.2 <0.2 <0.2 Antimony (Sb) Μg / L - 0.5 ± 0.2 1.3 ± 0.5 0.4 ± 0.3 Uranium (U) Μg / L - <0.2 0.4 ± 0.3 0.5 ± 0.5 Titanium (Ti) Μg / L - <0.2 1.0 ± 0.3 <0.2 Cobalt (Co) Μg / L - <0.2 <0.2 <0.2 Germanium (Ge) Μg / L - <0.2 <0.2 <0.2 Tin (Sn) Μg / L - <0.2 4.0 ± 0.5 <0.2 Zirconium (Zr) Μg / L - 0.4 ± 0.3 0.7 ± 0.5 0.5 ± 0.5 Tungsten (W) Μg / L - 0.4 ± 0.3 6.5 ± 1.0 1.9 ± 0.5 Cesium (Cs) Μg / L - <0.2 <0.2 <0.2

Reference Example 1 Preparation of Experimental Materials

Cell culture reagents such as RPMI-1640, modified Eagle Medium (MEM), non-essential amino acids solution, fetal bovine serum (FBS), streptomycin, penicillin, (Grand Island, USA). Among the reagents used in the experiment, c-kit ligand (KL) was obtained from STEMCELL Technologies (vancouver, Canada), lipopolysaccharide (lipopolysaccharide (LPS), interleukin (IL) -10, PBS, etc. were manufactured by Sigma (St. Louis, USA). EIA kits for prostaglandin D ₂ and leukotriene C ₄ were purchased from Cayman (Ann Arbor, USA).

Reference Example 2 Experimental Cell Culture

Mouse bone marrow-derived mast cells (BMMC) were isolated from bone marrow from BALB / C mice, and RPMI-1640 medium containing 10% FBS, 100 U / ml penicillin, 100 μg / ml streptomycin and IL -3 [cultured supernatant obtained by stimulating with pokeweed mitogen to mouse spleen cells to give a final 20%] was incubated for about 3 weeks to obtain more than 95% homogeneous BMMC.

Experimental Example 1 Inhibition of Mast Cell Degranulation Response

After treatment with a certain concentration of Jeju water to ₂ × 10 ⁵ cells / ml of mouse bone marrow-derived mast cells (BMMC) prepared in Reference Example 2, and pre-incubated for 30 minutes at 37 ℃, 5% CO ₂ conditions, Stimulation with KL (100 ng / ml) was incubated for 15 minutes and centrifuged for 5 minutes at 3000 rpm, 4 ℃.

The supernatant was beta-hexosaminidase (β-HEX) [100 mM citric acid buffer solution (0.955% citric acid, 1.478% sodium citrate dihydrate, pH 4.5), 1.3 mg / ml p-nitrophenyl-N -Acetyl-bD-glucosaminide] 1: 2 and reacted for 1 hour at 37 ℃, the reaction was stopped with 0.2M glycine (pH 10.7) to measure the absorbance at 405nm wavelength using ELISA The measured value was converted into% release granules.

[Equation 1]

% Degranulation = [free β-hex activity in culture supernatant / (released β-hex activity in culture supernatant + intracellular β-hex activity)] X 100

The results are shown in Figure 1 (a: Samdasoo, b: S2 and c: S3), the degree of inhibition is shown in Table 5 below as a percentage. Jeju water of the present invention showed a β-hexos aminidase free inhibitory effect in a concentration-dependent manner, as shown in Table 5 below, in particular, the best inhibitory effect in S3.

Jeju Water Throughput (μl) 10 20 40 Samdasoo 13.22 ± 0.5 17.54 ± 0.1 21.47 ± 0.7 S2 11.62 ± 0.7 19.25 ± 0.4 28.14 ± 0.5 S3 21.63 ± 1.5 30.88 ± 1.3 35.96 ± 1.0

<Experiment 2> Examination of the inhibitory effect of leukotriene production

In order to confirm the effect of Jeju water prepared in the above example on the production of leukotriene C ₄ , experiments were conducted by applying the method disclosed in the literature (Chang HW et al. Planta. Medica., 70 ( 5), pp. 474-476, 2004).

After 3 weeks of culture of mouse bone marrow-derived mast cells prepared in Reference Example 2, the pre-treated Jeju water prepared in the above example was pre-treated for 30 minutes at a predetermined concentration in mast cells (2Ｘ10 ⁵ cells / well), and then 100 Kng of stimulating agent was 100 ng / ml. The minutes were processed. The leukotriene C ₄ quantification of the supernatant after cell stimulation was measured using a leukotriene C ₄ EIA kit (Enzyme linked immunoassay, Cayman, product no. 520211).

The results are shown in FIG. 2 (a: samdasoo, b: S2 and c: S3), and the degree of inhibition is shown in Table 6 below as a percentage. Jeju water of the present invention showed the inhibitory effect of leukotriene C ₄ production in a concentration-dependent manner, as shown in Table 6 below, in particular, the best inhibitory effect in S3.

Jeju Water Throughput (μl) 10 20 40 Samdasoo 11.67 ± 1.1 16.07 ± 0.7 21.91 ± 0.5 S2 14.23 ± 0.9 24.53 ± 1.1 32.01 ± 0.5 S3 14.24 ± 1.5 26.07 ± 0.9 46.30 ± 1.2

Experimental Example 3 Cyclooxygenase-2-Dependent Prostaglandin D ₂  Review the effects of inhibition

Production amount of prostaglandin D ₂ (PGD ₂ ) using prostaglandin D ₂ EIA kit (Cayman, PGD2-MOX EIA kit, product no. 512011) to confirm the degree of inhibition of various inflammatory diseases of Jeju water prepared in Example Was measured.

After 3 weeks of culture of mouse bone marrow-derived mast cells (BMMC) prepared according to Reference Example 2, after pretreatment of Jeju water prepared in the above example for 30 minutes at a cell concentration of 2Ｘ10 ⁵ cells / well for 30 minutes, KL of 100ng / ml (c-kit ligand, mast cell proliferation factor), 100 ng / ml of LPS (Mast Cell Activator) and 100 U / ml of IL-10 (Mast Cell Activator) After time, the prostaglandin D ₂ of the supernatant was measured by an Enzyme linked immunoassay (EIA) using the prostaglandin D ₂ assay kit (Cayman). At this time, in order to inhibit the production of prostaglandin D ₂ produced by cyclooxygenase-1, 10 μg / ml of aspirin was pretreated with mouse bone marrow-derived mast cells one hour beforehand.

After washing 4 times with a washing buffer, the substrate solution was treated with 200 μl each for 5-20 minutes, and then treated with 50 μl stop solution, followed by absorbance at 450 nm. Was measured by ELx800 (BIO-TEK, Instrument, Inc, USA).

The results are shown in FIG. 3 (a: samdasoo, b: S2 and c: S3), and the degree of inhibition is shown in Table 7 below as a percentage. Jeju water of the present invention showed a cyclooxygenase-2 dependent prostaglandin D ₂ production inhibitory effect in a concentration-dependent manner, as shown in Table 7 below, in particular the best inhibitory effect in S3.

Jeju Water Throughput (μl) 10 20 40 Samdasoo -3.67 ± 0.8 -6.05 ± 3.5 5.80 ± 0.6 S2 4.39 ± 1.0 6.66 ± 0.8 12.44 ± 0.2 S3 7.60 ± 0.6 11.73 ± 0.3 22.40 ± 0.3

<Experimental Example 4> Toxicity test

Jeju water (Samdasoo, S2 and S3) prepared in Example was once orally administered to male Balb / c mice at doses of 0.1 mL / kg, 0.5 mL / kg and 1 mL / kg, and the survival rate and body weight of the mouse for 7 days Was investigated.

After this administration, we observed the mortality of the animal, clinical symptoms, weight change, hematologic test and blood biochemical test, and autopsied the visceral organs and thoracic organs were observed.

As a result, no clinical symptoms or dead animals were found in all animals, and no toxic changes were observed in weight changes, blood tests, blood biochemical tests, and autopsy findings.

As a result, the Jeju water (Samdasoo, S2 and S3) of the present invention does not show a toxic change up to 1 mL / kg in the mouse, therefore, oral administration medium dose (LD50) was determined to be a safe substance of 1 mL / kg or more.

Hereinafter, a preparation example including Jeju water S3 according to the present invention will be described, but the present invention is only intended to be described in detail and not intended to limit the same.

&Lt; Formulation Example 1 > Preparation of health food

Vitamin Mixture (70 μg Vitamin A Acetate, 1.0 mg Vitamin E, 0.13 mg Vitamin B 1, 0.15 mg Vitamin B 2, 0.5 mg Vitamin B 6, 0.2 μg Vitamin B 12, Vitamin C 10 mg, Biotin 10 μg, Nicotinamide) 1.7 mg, folic acid 50 µg, calcium pantothenate 0.5 mg, mineral mixture (1.75 mg ferrous sulfate, 0.82 mg zinc oxide, magnesium 25.3 mg, potassium monophosphate 15 mg, dibasic calcium phosphate 55 mg, potassium citrate) 90 mg, calcium carbonate 100 mg, magnesium chloride 24.8 mg) and Jeju Water S3 were added to make the total 900 mL, and then the health food was prepared according to a conventional method using the concentrate obtained by concentrating.

Preparation Example 2 Preparation of Health Beverage

1000 mg of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, 1 g of taurine, and Jeju water S3 were added to make 900 ml. After mixing the above components according to the usual method of preparing healthy drinks, the mixture was 85 for about 1 hour. After stirring and heating at ° C., the resulting solution was collected by filtration into a sterilized 2 L container, sealed sterilized and refrigerated.

Preparation Example 3 Preparation of Powder

100 mg of lactose, 10 mg of talc and Jeju Water S3 were added to make it total 900 ml, and the concentrated concentrate was filled into an airtight cloth to prepare a powder.

Preparation Example 4 Preparation of Tablet

100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate, and Jeju water S3 were added and mixed to a total amount of 900 ml, and the concentrated concentrate was compressed into tablets according to a conventional method for preparing tablets.

Preparation Example 5 Preparation of Capsule

100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate, and Jeju water S3 were added and mixed to a total amount of 900 ml. Then, the concentrated concentrate was mixed according to a conventional capsule preparation method and mixed into gelatin capsules. The capsule was prepared by filling.

Claims (8)

7 to 30 ppb (μg / L) of vanadium and 15 to 50 ppb (μg / L) of excavated from 200 to 600 m underground in any one or more areas selected from Gyorae-ri, Jocheon-eup, Jeju-si, Jungmun-dong, Seogwipo-si and Dosun-dong, Seogwipo-si Allergic disease, inflammatory bowel disease, systemic lupus erythematosus, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, pancreatitis and nephritis containing Jeju water containing strontium as an active ingredient Pharmaceutical composition for preventing or treating any one inflammatory disease selected from the group consisting of.
delete delete delete delete The allergic disease of claim 1, wherein the allergic disease is any one selected from the group consisting of hypersensitivity, allergic rhinitis, asthma, allergic conjunctivitis, allergic dermatitis, atopic dermatitis, contact dermatitis and urticaria. Inflammatory bowel disease, systemic lupus erythematosus, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, pancreatitis and nephritis Composition. 7 to 30 ppb (μg / L) of vanadium and 15 to 50 ppb (μg / L) of excavated from 200 to 600 m underground in any one or more areas selected from Gyorae-ri, Jocheon-eup, Jeju-si, Jungmun-dong, Seogwipo-si and Dosun-dong, Seogwipo-si Allergic disease, inflammatory bowel disease, systemic lupus erythematosus, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, pancreatitis and nephritis containing Jeju water containing strontium as an active ingredient Functional food composition for preventing or improving any one inflammatory disease selected from the group consisting of. The method of claim 7, wherein the functional food is an allergic disease, inflammatory bowel disease, systemic lupus erythematosus, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis , Pancreatitis and nephritis functional food composition for preventing or improving any one inflammatory disease selected from the group consisting of.

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