KR20030056582A - Composition for immune enhancement - Google Patents

Abstract

PURPOSE: An immunostimulating composition containing a natural herbal extract increasing effect on the production of interleukin-2 and interferon-γ and inhibiting secretion of histamine is provided. Therefore, the composition can be effectively used as an agent for or an adjunct to treatment of diseases according to the lowering of immunity, in particular, cold. CONSTITUTION: An immunostimulating composition contains an extract of Glycyrrhizae Radix, Zizyphi Fructus, Pyrus pyrifolia(Burm. fil.) Nakai var. culta (Makino) Nakai, Atractylodis rhizoma alba, Zingiberis Rhizoma, Folium Perillae, Astragali Radix and green onion, Allium fistulosum L. in a weight ratio of 1 to 4:1 to 4: to 10 to 40:3 to 9:1 to 4:1 to 4:2 to 6:5 to 10. The herbal extract is added with a mixture of rice, malted rice and yeast in a ratio of 1 to 4:0.2 to 1:0 to 0.1. and then fermented to produce the immunostimulating composition.

Description

Composition for immune enhancement

The present invention relates to an immune enhancing composition having effects such as cold prevention. More specifically, the present invention relates to a composition for enhancing immunity containing extracts of licorice, jujube, pear, baekchul, ginger, lobule, dermis, and baekbaek as an active ingredient.

Colds are the most frequently treated diseases in primary health care. About a third to a quarter of patients who visit general hospitals are acute upper respiratory tract infections called colds. Pediatric airway infections are particularly serious and can be life threatening. In children under five, airway infections are known to be the most important cause of death from lung disease. Infectious diseases such as these play an important role in immune function. Therefore, increased immunity can be a good way to prevent and treat colds.

In recent years, the market for alternative medicine for the prevention and treatment of colds is growing rapidly. In the United States, about one-third of the population relies on alternative medicines, not vaccines and chemicals. . In light of this trend, there is an urgent need for the development of a medicine or a food composition having an effect of increasing immune function as an alternative medicine such as herbal medicine.

The immune system can be divided into natural resistance, nonspecific immune system and specific immune system. Natural resistance (primary defense) refers to anatomical physiological elements that block all invaders, including microorganisms, regardless of their type. Nonspecific immunity (secondary defense) is a phagocyte that breaks through natural resistance and removes invaders. The constructed defense system, and the specific immune system (tertiary line of defense) refer to the immune system composed of lymphocytes, which are highly developed immune systems with memory and the ability to distinguish between self and nonmagnetic.

Leukocytes constitute a secondary or tertiary line of defense, breaking through the primary line of defense, and are responsible for foreign bodies entering the body. Macrophages contain many lysosomes, which contain acidic hydrolase and peroxidase. In addition, it strongly adheres to the glass surface and plastic surface, and actively detects microorganisms and tumor cells. This cell has cytokine receptors such as IFN-γ. They produce cytokines such as complement, interferon, interleukin-1 and tumor necrosis factor and can be enhanced by several cytokines produced from T-cells.

T-cells, also a type of white blood cell, make up about 70% of blood lymphocytes. T-cells differentiate in the thymus and have a T-cell antigen receptor (TCR). Peripheral blood T- cell is divided into CD4-positive T _H cells (helper T-cell) and CD8-positive TC cells (cytotoxic T-cell). CD4 ⁺ T _H cells are activated by recognizing antigens bound to MHC class II molecules, helping B cells to produce antibodies or to help other T cells function. CD4 ⁺ T _H cells can again be classified into T _H 1 and T _H 2 based on the cytokines they produce. T _H 1 cells in mice secrete interleukin-2 (IL-2), interferon-gamma (IFN-γ), etc., whereas T _H 2 cells produce IL-4, IL-5, IL-6 Secrete IL-9, IL-10, IL-13 and the like. However, in humans, the production of IL-2, IL-6, IL-10 and IL-13 is not strictly distinguished. In addition, IL-3, tumor necrosis factor-alpha (TNF-α), granulocyte-macrophage colony-stimulating factor (GM-CSF), (meth) enkephalin (chem) and chemokine (CK) are also distinguished. Are commonly secreted. T _H 1 cells are involved in cellular immune responses and activate cytotoxic and inflammatory responses. Cytokines produced in T _H 2 cells promote antibody formation, particularly the production of IgE and enhance the proliferation and function of eosinophils. Therefore T _H 2 cytokines are commonly found in antibody production and allergic reactions. It has been shown that T _H 1 and T _H 2 cytokines have mutually inhibitory functions and that the disease progression can be altered with anti-IL-4 and anti-IFN-γ antibodies in animal infections. In some cases, improvement of symptoms was observed by injecting IFN-γ.

As described above, substances having excellent production ability such as interleukin-2 (IL-2) and interferon-gamma (IFN-γ) are expected to have an immune enhancing effect, and further help in preventing cold or treating early cold. It is expected to be.

An object of the present invention is to identify the antihistamine effect, IL-2 and interferon-gamma-producing ability of pharmaceutically acceptable compositions containing natural products with no side effects, thereby enhancing immunity such as cold prevention and early cold treatment effects. It is to provide a composition for.

Figure 1 shows the effect of increasing interferon-gamma production for the composition according to the present invention,

Figure 2 shows the histamine secretion inhibitory effect on mast cells for the composition according to the present invention.

The present invention provides an immune enhancing composition containing extracts of licorice, jujube, pear, baekchul, ginger, lobule, dermis and baekbaek.

As the extraction solvent, water or a conventional organic solvent such as methanol, ethanol, propanol, and the like may be used, and preferably water is used. In addition, sweeteners such as brown sugar may be added to improve taste.

The disease in which the composition of the present invention is effective may be, in particular, weak constitution, lowering immunity, initial cold, and the like.

In the composition according to the present invention, licorice, jujube, pear, baekchul, ginger, lobule, dermis, and ginseng are 1-4: 1-4: 10-40: 3-9: 1-4: 1-4: 2-6 It is preferable to mix at a weight ratio of 5-10. In addition to the above composition, brown sugar may be mixed in a weight ratio of 1 to 4.

Hereinafter, the present invention will be described in detail.

Licorice ( Glycyrrhiza uralensis FISCH), the first active ingredient of the composition according to the present invention, is a perennial herb and muscles of the licorice. This product plays the role of Bobi ripening, clear heat detoxification, decontamination, harmonious pharmaceuticals. This product harmonizes several drugs to relieve the mild properties of the drug.

Jujube ( Zizyphus jujuba Mill. Var inermis (Bunge) Rehd), the second active ingredient of the composition according to the present invention, is a mature fruit of the jujube tree, which is a deciduous shrub or sub-tree. This product has the effects of Bobihwawi, pterosaur, anxin.

Pyrus pyrifolia var. Culta (Makino) Nakai, the third active ingredient of the composition according to the present invention, is a mature fruit of a deciduous arborescent or shrub plant belonging to the Rosaceae family. This product is an inorganic ingredient that contains 74% of calcium, sodium, and magnesium, and 25% of phosphorus and lactic acid, which is a strong alkaline food, so eating a lot of pears or processed foods keeps our blood neutral to maintain health. It has a big effect. The type of vitamins contained in the pear is not much higher than other fruits, but the vitamin B1 and B2 content is slightly higher than the apples, and the vitamin C content is low. Vitamin C is made from glucose produced by assimilation in leaves, which is known to accumulate in the fruit. In general, the amount of vitamin C is higher in the sunlight, which affects the production of vitamin C. . It is effective to use sputum with severe asthma, coughing of young children and whooping cough. The nature of the belly is cold, but effective in digestion, the feces or urine will come out to make the body heat. In the case of severe thirst or severe hangovers, the stomach stimulates liver activity to quickly detoxify the alcohol in the body, so the poison is quenched and thirst is eliminated.

Atractylodes macrocephala Koidz, the fourth active ingredient of the composition according to the present invention, is a perennial herb perennial herb of the Asteraceae family and the root of the same plant. This product has the effects of Bobi ripening, humidity control, high labeling. Applied to symptoms such as obesity caused by rectification of probation. In addition, since it has a high labeling effect, it can be applied to the deadline due to the license.

Ginger ( Zingiber offininale Rosc), the fifth active ingredient of the composition according to the present invention, is a fresh root of ginger, which is a perennial herb belonging to the family Ginger. This article has the effect of sweat sweating, warm earth district, the warming of the sea. It is used for the increase of feelings of admiration and the unauthorized.

The sixth active ingredient ( Perilla frutescens (L.) var. Acuta (Thunb.) Kudo) of the composition according to the present invention is a leaf or an outpost of perennial herb, which is a herbaceous herb belonging to the genus. This article has the efficacy of sweat sweating, affairs, seafish poison. It is utilized for indications of sensation of wind, symptom of seawater rash.

Citrus unshiu Marcor, the seventh active ingredient of the composition according to the present invention, is a mature rind of the evergreen cultivars of tangerine and related plants. This product has the effects of ego-gun dry, moist painting. Treats gastrointestinal gastropathy or gastrointestinal symptoms.

The eighth active ingredient (Allium fistulosum L.) of the composition according to the present invention is a perennial herb, which belongs to the family Liliaceae. This article has efficacy such as tonalization and expression. Headache caused by cold, cold, fever, etc.

In addition, black sugar (black sugar) that can be added to the composition according to the present invention to improve the taste is one of the sugar-containing sugar produced in sugar cane. The molasses of sugar cane is boiled and the molasses is not removed by centrifugal separator. The juice from the stem of sugar cane is added to a juice solution, such as lime, and then boiled. Brown sugar is not only rich in minerals and vitamins that are lost in the refining stage, but also effective in the body.

In the composition for enhancing immunity according to the present invention, each herbal medicine is extracted by the conventional method, respectively, or after mixing the herbal medicines at once, and if necessary, concentrated and dried to powder them. In the immune enhancing composition according to the present invention, each herbal medicine is preferably obtained by mixing with a prescribed weight ratio and then extracting with water.

The immune enhancing composition according to the present invention preferably further contains a pharmaceutically acceptable carrier. Carriers that can be used in the composition of the present invention are conventional in the pharmaceutical field, for example, in the case of oral preparations, there are binders, lubricants, disintegrants, excipients, solubilizers, stabilizers, and the like. In the case of a solvent preparation, there are a base, an excipient, a lubricant, and a preservative.

Compositions according to the present invention are conventional formulations in the pharmaceutical field, such as tablets, capsules, troches, solutions, suspensions or the like, oral injectable solutions or suspensions, or distilled water for injection upon injection. It may be formulated into a variety of preparations, such as topical preparations such as injectable preparations, ointments, creams, liquids, etc. in the form of ready-to-use injectable dry powders that can be prepared and used. The composition according to the invention is preferably in liquid form. The composition according to the present invention is more preferably in the form of a subject made by alcohol fermentation by adding fermentation raw materials to extracts of the active ingredients as herbal ingredients. As the fermentation raw material, any grains commonly used for alcoholic fermentation can be used, for example, rice, yeast, yeast, glutinous rice, crude and the like. As the fermentation raw material, it is particularly preferable to use a mixture of rice, yeast and yeast in view of taste and simplicity.

In a preferred embodiment of the present invention, the composition according to the present invention has licorice, jujube, pear, baekchul, ginger, lobule, dermis, and baekbaek 1-4: 1-4: 10-40: 3-9: 1-4: It contains in the weight ratio of 1-4: 2-6: 5-10. In addition, it is preferable to further contain rice, yeast and yeast and to ferment by a conventional method to produce a functional concentration of a suitable concentration. That is, licorice, jujube, pear, white peach, ginger, lobule, dermis and total white are 1-4: 1-4: 10-40: 3-9: 1-4: 1-4: 2-6: 5-10 To the water extract obtained by mixing in a weight ratio and decanted, rice, yeast and yeast were mixed at a weight ratio of 100 to 400: 20 to 100: 0 to 10 to 30 to 40% by weight of the water extract and fermented to a suitable concentration. Manufactured with the theme of.

In addition, the composition of the present invention is licorice, jujube, pear, baekchul, ginger, leaflet, dermis and total white 1-4: 1-4: 10-40: 3-9: 1-4: 1-4: 2-6: Also included is an alcohol extract obtained by mixing and fermenting a mixture of 5 to 10 in a weight ratio of rice: 100 to 400 in a ratio of yeast 20 to 100: yeast 0 to 10.

The composition ratios of the above-mentioned herbal medicines are obtained based on the repeated experimental results, and when the composition ratio of each herbal medicine is lower than the lower limit, the pharmacological effect of the composition of the present invention is reduced, and when higher than the upper limit, the pharmacological effect of each component Can be reduced so that the synergy and coordination of the composition of the present invention is reduced.

The composition of the present invention can be used as an adjuvant for preventing or treating colds through enhanced immunity, and such pharmacological effects are confirmed from Experimental Examples 1 to 3 below. The composition according to the present invention is estimated to enhance the local or systemic immune capacity by regulating the production of cytokines and chemical mediators from immune cells.

Preferred examples of the composition of the present invention include licorice 10-40g, jujube 10-40g, pear 100-40g, white 100-90g, ginger 10-40g, leaflet 10-40g, dermis 20-60g, total white 50-100g and Pour 6,000 ml of water into 10 to 40 g of brown sugar, decoction for about 2.5 hours, concentrate to about 4,000 ml to obtain an extract, and mix uniformly the steamed yeast, 0.2-1 kg of yeast and 0-0.1 kg of yeast, which are made separately from 1-4 kg of rice The extract was poured into the solution, fermented at 20 to 27 ° C. for 5 to 10 days, filtered to prepare a subject, and then purified water was adjusted to 3,000 ml.

A typical single dose of the composition is 0.1-5.0 ml, preferably 1.0-2.0 ml per kg body weight, which is taken three times a day. For example, for adults weighing 60 kg, take 6 to 300 ml once three times a day. When taken at this dose, the preparation is about 8 to 15 days old. However, the composition of the present invention can be appropriately increased or decreased the dose depending on the weight, age, sex, severity of the patient and the digestive state. In other formulations, an appropriate amount calculated in consideration of the liquid dosage can be administered orally or parenterally. As early as two to three days after taking the symptoms of the symptoms can be reduced, but there are individual differences. Take it for about a month after the effect starts.

Hereinafter, the present invention will be described in more detail with reference to the following Examples. The scope of the present invention is not limited in any way.

Example 1 Preparation of Liquid

Licorice 4g, jujube 4g, pear 40g, baekchul 6g, ginger 4g, lobule 4g, dermis 6g and a total of 10g poured 1,000ml of water, decoction for about 2 hours, concentrated to about 150ml to prepare a liquid formulation.

Example 2: Preparation of Moisture

Licorice 40g, jujube 80g, pear 800g, white roe 120g, ginger 80g, lobule 40g, dermis 60g, total white 200g, brown sugar 80g pour 6,000 ml of water, decoction for about 2.5 hours, concentrated to about 4,000 ml to obtain an extract To the extract, a mixture of steamed rice, yeast 1 kg and yeast 50 g, which was separately made from 2 kg of rice, was poured into 30% by weight of the extract and fermented at 20 to 27 ° C. for 7 days. The fermentation broth was filtered to separate the liquid and solid portions, heated to 55-60 ° C., and purified water was added to the filtrate.

Experimental Example 1: Toxicity and Side Effects Identification Test

1 times (2 mL / kg), 5 times (10 mL / kg), and 10 times the dose ratio (dose of 1-2 ml per kg of body weight) of administering the liquid preparation prepared in Example 1 to human 20 ml / kg) was administered orally to 30 adult rats for 90 days and then observed daily for more than 30 days. As a result, no abnormalities were found in all experimental groups as well as in physiological abnormalities.

Accordingly, the present inventors studied the appropriate dosage and administration method of the liquid formulation for adult male and female volunteers after 20 years of age. Except for the appearance of gastrointestinal discomfort, transient phenomena or intoxication in a small number of people, no serious local or systemic side effects have been identified, and the gastrointestinal discomfort level is 1/2 to 2/3. After losing weight, it disappeared remarkably.

Experimental Example 2 effect of increasing interferon-gamma production

Interferon-gamma is a cytokine secreted by T _H 1 cells and plays an important role in immune function and its increase can be understood as an indicator of increased immune function. Malt-4, a cell line of T _H 1 cells, was cultured and seeded at 2 × 10 ⁵ cell / well and stabilized for 30 minutes. Samples powdered by lyophilizing the subject of Example 2 (concentrations of 0.01 mg / ml, 0.1 mg / ml, 1.0 mg / ml) and other stimulants, such as Phytohaemaglutinin (PHA) After 20 hr / ml treatment, the production of interferon-gamma was measured using the ELISA method after 24 hours, and the results are shown in FIG. 1.

As shown in Figure 1, the composition of the present invention increased the amount of interferon-gamma produced in a concentration-dependent manner.

Experimental Example 3: Measurement of the production capacity of interleukin-2

In addition to interferon-gamma, interleukin-2 also plays an important role in immune function as a cytokine secreted by T _H 1 cells, and its increase can also be understood as an indicator of increased immune function. Malt-4, a cell line of T _H 1 cells, was cultured and seeded with 2 × 10 ⁵ cell / well and stabilized for 30 minutes. The subject of Example 2 was lyophilized and treated with powdered samples (0.01 mg / ml, 0.1 mg / ml, 1.0 mg / ml), and the production of interleukin-2 was determined after 24 hours using the ELISA method. And the results are shown in Table 1.

Concentration of Compositions of the Invention (mg / ml) IL-2 (ng / ml) 0 0.103 ± 0.01 0.1 0.231 ± 0.04 One 0.354 ± 0.07 ^*

As shown in Table 1, the composition of the present invention significantly increased the amount of interleukin-2 produced.

Experimental Example 4: Histamine secretion inhibitory effect

One of the most common cold symptoms is an allergic reaction, such as rhinitis or coloritis. Steroidal antihistamines are widely used as a symptomatic treatment for these symptoms. However, these antihistamines have many side effects and their effects are temporary. Therefore, the inhibitory effect of histamine secretion as an adjuvant against colds and treatment through immunity can be an important indicator. Therefore, the histamine inhibitory effect on the mast cell line was observed with the composition of the present invention. Rats were anesthetized with ether, followed by Tyrode buffer B (1 mM MgCl ₂ , 1.8 mM CaCl ₂ , 137 mM NaCl, 12 mM NaHCO ₃ , 2.7 mM KCl, 0.2 mM NaH ₂ PO ₄ ) containing 0.1% gelatin. , 5.5mM glucose) was injected intraperitoneally, and the abdominal wall was lightly massaged for 30 seconds. Then, the abdominal wall center line was carefully cut and a washing solution containing peritoneal cells was collected with a Pasteur pipette. The peritoneal cells were centrifuged at 150 × g for 3 min each time, and the supernatant was discarded and resuspended with the same T-load buffer B. The mast cells in the cell suspension were separated and purified using 22.5% (w / v) metrizamide. In other words, 22.5% (w / v) methazamide (concentration 1.120 g / ml) was superimposed on peritoneal cells (6 to 9 x 10 ⁷ ) suspended in 1 ml of T-load buffer B, and then heated at 400 x g for 15 minutes. Centrifugation at. Cells at the buffer and methazamide contact surfaces were collected and resuspended in pellets of 1 ml of T-load buffer B. This procedure was repeated to obtain peritoneal mast cells with greater than 99% purity.

The mast cells thus obtained were added to 96 wells, and three samples, each of which was powdered with a subject according to Example 2, were added to each well at concentrations of 0.01, 0.1 and 1 mg / ml. Distilled water was added as a control. Then, the amount of compound 48/80 6 ㎍ / ml was added to each well to induce histamine secretion. After 20 minutes at 37 ° C., the amount of histamine released was measured. In order to measure the amount of histamine, the following procedure was carried out.

Centrifuge the sample in each well for 5 minutes at 3000 rpm, and then put 500 μl of the supernatant into the eppendorf tube, add 450 μl of 0.1N-HCI, 50 μl of 60% perchloric acid solution, and mix and centrifuge (1,500 rpm, 20 minutes). The supernatant was added to 800 µl of a 5 N-NaOH solution, 3 ml of distilled water, 10 ml of n-butanol, and 1.2 g of NaCl, followed by centrifugation (2,000 rpm, 10 minutes) after shaking. 8 ml of butanol layer was placed in a 50 ml test tube, and 3 ml of 0.1 N-HCI solution and 10 ml of n-heptane were added thereto, followed by centrifugation (2,000 rpm, 10 minutes). To the aqueous layer obtained here with 2 ml 1 N-NaOH solution 400 ㎕ 1% o - au taldi aldehyde (o -phthaldialdehyde) into a solution of 100 ㎕, allowed to stand for two minutes after mixing the fluorine-meter spectrometer (spectrofluormeter) (λex = 360nm , λex = 440 nm) was measured to measure the amount of histamine secreted, and the results are shown in FIG.

As shown in Figure 2, the composition of the present invention was excellent in histamine inhibitory effect.

As described above, it can be seen that the composition of the present invention has an increase in interleukin-2 and interferon-gamma production. The ability to produce interleukin-2 and interferon-gamma (IFN- [gamma]) is directly or indirectly related to the immune enhancing effect. In addition, it can be seen that the composition of the present invention also has an effect of inhibiting the release of histamine. Among the many positive functions that can be expected from immune-boosting effects include the prevention of colds. The inhibitory effect of histamine secretion may play a role in alleviating the symptoms of early cold.

Therefore, the composition of the present invention can be usefully used in the prevention of diseases caused by lowering immunity, especially cold and treatment or treatment aid of early cold in modern industrial society in which immunity is lowered by exposure to various harmful environments.

Claims (7)

Immunity-enhancing composition containing licorice, jujube, pear, baekchul, ginger, lobule, dermis and baekbaek extract. The composition of claim 1, wherein the extract is a water extract obtained by extraction with water. The licorice, jujube, pear, white extract, ginger, lobule, dermis and total white are 1-4: 1-4: 10-40: 3-9: 1-4: 1-4: A composition mixed at a weight ratio of 2 to 6: 5 to 10. The composition of claim 1 or 2, further comprising brown sugar. The composition according to claim 2, wherein the composition is in the form of a main ingredient obtained by adding fermentation raw material to the water extract and fermenting the same. The composition of claim 5, wherein the fermentation raw material is a mixture of rice, yeast and yeast mixed in a weight ratio of 1 to 4: 0.2 to 1: 0 to 0.1. A method of producing the composition of claim 5, wherein a mixture of rice, yeast and yeast is added to the water extracts of licorice, jujube, pear, baekchul, ginger, lobule, dermis and ginseng mixture.