KR20050014615A - Composition comprising Schizandrae Fructus extract as an effective component for preventing and treating arthritis - Google Patents

Abstract

PURPOSE: A composition for treatment and prevention of osteoarthritis containing Fructus schizandrae extract having an action of inhibiting hyaluronidase is provided. It inhibits hyaluronic acid decomposition by hyaluronidase in a joint generally produced in osteoarthritis patients and the inhibition of an extracellular matrix of chondrocyte cells and therefore can be used as a drug and health food for prevention and treatment of arthritis. CONSTITUTION: The composition for treatment and prevention of osteoarthritis contains Fructus schizandrae extract as an effective component. The Fructus schizandrae extract is extracted by the steps of: extracting with methanol; pouring distilled water into the methanol extract and extracting with hexane; reextracting the extraction residue with chloroform; and extracting the extraction residue with ethyl acetate. The daily dose of the composition is 1 to 100mg/kg. For an example, 100mg Fructus schizandrae extract is mixed with 100mg corn starch, 100mg lactose and 2mg magnesium stearate to give a tablet.

Description

Composition comprising Schizandrae Fructus extract as an effective component for preventing and treating arthritis

The present invention relates to a composition for preventing and treating arthritis containing Schizandra chinensis extract as an active ingredient.

Arthritis is a representative of cartilage-related diseases, which are roughly classified into rheumatoid arthritis and degenerative arthritis, and knee joint cartilage is often found in traffic accidents or excessive exercise. At present, there are about one million arthritis patients in Korea, and women are more than twice as many men, especially menopausal women.

Rheumatoid arthritis is a type of autoimmune disease, and it occurs in women of 30 to 50 years at a ratio of 1: 4. Symptoms include symmetrical inflammation of the joints of fingers, elbows, knees, etc. It is severe at dawn, weakness and mild fever. Arthritis of acute joint rheumatism disappears within a few days to weeks, whereas rheumatoid arthritis lasts for months and years, resulting in joint deformation, dislocation, muscle and tendon buildup. And stiffness may cause severe disability.

Degenerative arthritis is a disease in which local degenerative changes occur as the articular cartilage wears off. Also called osteoarthritis or osteoarthritis. Osteoarthritis is a representative degenerative disease that is closely related to aging. About 10 to 15% of the entire population suffers from osteoarthritis. In particular, about 60 to 80% of the elderly aged 65 or older suffer from osteoarthritis.

The causes of degenerative arthritis are deeply related to senility and excessive weight, and the more and more severely in women as they age. The initial symptom is that one or two joints are swelling with stiffness, and if prolonged, the joints are deformed.

The cause of degenerative changes in articular cartilage has not yet been identified, but an absolute decrease in the number of chondrocytes and imbalances in the synthesis and degradation of chondrocytes in chondrocytes is known as one cause. Thus, degenerative changes in articular cartilage reduce cartilage strength and ability as a cushion due to degradation of cartilage substrate.

Currently, clinically used degenerative arthritis can be treated with medications (painkillers, steroids, nonsteroidal anti-inflammatory drugs), cartilage protectors (hyaluronic acid, glucosamine, chondroitin, etc.) or surgical treatment (arthroscopy, proximal tibia). Osteotomy, joint replacement, total knee arthroplasty, etc.). However, drug therapy has only the effect of non-specifically relieving pain or inflammatory response itself, and cartilage protection agent only serves to protect joints by nourishing chondrocytes or alleviating shock. In addition, steroidal drugs have side effects that can lead to osteoporosis, high blood pressure, diabetes, etc. due to loss of calcium when taken for a long time. Therefore, in most cases, the drug is used only for the purpose of reducing pain, and permanent arthroplasty is mainly used, but there are no drugs or surgical methods that provide a fundamental therapeutic effect.

Although much research and efforts have been made for the treatment of degenerative arthritis in the past decades, the pathological cause and the underlying treatment are not yet developed.

Recently, the treatment of arthritis through herbal and nutritional supplements has been developed, but the therapeutic effect and mechanism of action are not known. Therefore, these arthritis diseases are not only difficult to effectively control the disease with the simple anti-inflammatory drugs, the existing simple symptomatic therapy, but also cause a lot of difficulties in the treatment of the elderly, which is the majority of patients in reality due to the side effects of long-term administration of the drug.

Therefore, in order to treat arthritis, since it is necessary to take a medicament for a long time, development of a medicament with few side effects is desired.

Accordingly, the present inventors completed the present invention while searching for a substance for inhibiting hyaluronicides among herbal extracts, and there was a substance for inhibiting hyaluronicase in Schizandra chinensis extract.

The present invention confirms that Schisandra chinensis extract has an action of inhibiting hyaluronides, and is intended to be used in medicine or food for preventing and treating arthritis.

1 is a schematic diagram showing a method for extracting, separating and purifying Schizandra chinensis extract according to the present invention.

2 is a diagram showing the effect of Schizandra extract of the present invention on chondrocyte collagen type II expression.

The present invention provides a composition for the prevention and treatment of arthritis, containing Schizandra chinensis extract as an active ingredient.

The present invention also provides a hyaluronidate inhibitor containing Schizandra chinensis extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

Omija's scientific name is Schizandra chinensis Baillon, a deciduous broad-leaved vine whose leaves are alternate, ovate or obovate, with sharply pointed, jagged, somewhat hairy on the back veins. Flower is reddish white and blooms in June ~ July. Fruits are used for food and hang on the ends of branches. They ripen red in August-September. Fruits are called Schisandra (Fructus) and Schizandrae Fructus. 200 ~ 1,600m above sea level Jirisan, Jeonbuk, and Gangwon-do are the main regions, and are wild in the whole country except Chungnam and Chungbuk, and geographically distributed in Japan, Sakhalin, Manchuria, and China. Fruits include chizandrol, chizandrin A, B, C, angeloylgomisin O, P, epigomisin, pregomoisin, deoxyschizandrin ), And the like. Schisandra chinensis has long been used in herbal medicine as a raw material of medicinal herbs and has long-standing effects such as astringent, nourishment, tonic, ginhae medicine, haejudok, thirsty, astringent shovel, ripe ginjin and bosin salt. As a result, human stability has been confirmed according to the intake of the body. Schisandra chinensis extract stimulates the central nervous system, improves tonic action, intellect and labor capacity, and affects glucose metabolism to promote catabolism of hepatic glycogen, increasing glycogen content, promoting fatigue recovery, controlling blood pressure, and secreting gastric juice. It is used for low blood pressure, arteriosclerosis, diabetes, hepatitis, etc., because it has anti-oxidant effect, antibiotic detoxification function, relieves eczema and inflammation of skin, inhibits skin aging, antibacterial and whitening effect and is also used in cosmetics.

Preparation method of Schizandra chinensis extract of the present invention is as follows.

Schizandra chinensis is extracted with water or organic solvent. The organic solvent is used by selecting from alcohol, chloroform, hexane, ethyl acetate.

The extraction method may be a conventional extraction method such as cold needle, warm needle, heating using the solvent.

Schizandra chinensis extract of the present invention inhibits the activity of excellent hyaluronides.

Hyaluronic acid (hyaluronic acid) is a substance that is contained in cartilage tissue in vivo, it plays a big role in increasing the elasticity of the cartilage tissue, has a high moisture content, and has excellent viscoelasticity.

Intraarticular injection of hyaluronic acid has been reported to reduce the pain caused by osteoarthritis and to treat it.

In addition, there is a research report that collagenase and hyaluronidase, etc. increase the cell death of articular chondrocytes and decrease the metabolism of chondrocytes, thereby promoting the degenerative changes of chondrocytes.

Since the decomposition of hyaluronic acid is one of the factors causing osteoarthritis or the process of promoting osteoarthritis, the inhibitor of hyaluronic acid degrading enzyme (hyaluronidase) can be used as a preventive or therapeutic agent for osteoarthritis.

Schizandra chinensis extract of the present invention exhibits an excellent hyaluronatease inhibitory effect.

In particular, methanol extract and ethyl acetate extract showed an excellent effect, n-hexane or chloroform extract has a weak effect of inhibiting hyaluronidase.

In the present invention, N-acetylglucosamine (N-acetylglucosamine) produced by decomposition by hyaluronidase is measured by colorimetric determination at 585 nm using DMAB (p-dimetylaminobenzaldehyde).

As described above, it can be seen that the Schizandra chinensis extract of the present invention exhibits an excellent hyaluronidate inhibitory effect.

In order to determine the effect of hyaluronidase inhibitory activity on the extracellular matrix synthesis of chondrocytes in the present invention, after incubating the hyaluronidase into chondrocytes and the treatment of hyaluronateides and Schisandra chinensis extract together to the chondrocytes The expression level of collagen type II, a specific extracellular matrix of chondrocytes, is measured.

As described above, the Schizandrae chinensis extract of the present invention inhibits hyaluronides inhibiting the extracellular matrix synthesis of chondrocytes.

The composition of the present invention may contain one or more active ingredients exhibiting the same or similar function in addition to the Schizandra chinensis extract.

The composition of the present invention may contain at least one active ingredient exhibiting another function in addition to the Schizandra chinensis extract.

The composition of the present invention, in addition to the above-described active ingredient for administration may further comprise one or more carriers which are acceptable in pharmaceutical or food. Pharmaceutical or food acceptable carriers may be used in combination with saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components, as necessary. And other conventional additives such as buffers and bacteriostatic agents can be added. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA.

The composition of the present invention can be parenterally or orally administered according to the desired method, oral administration is preferred. The dosage varies depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, excretion rate, and severity of the disease, but the daily dosage is 1-100 mg / kg once It is preferable to administer dividedly to several times.

The composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention and treatment of arthritis.

Examples of preparations for the compositions of the present invention are illustrated below.

Formulation Example 1 : Preparation of Pharmaceutical Formulations

1. Preparation of powder

Schisandra chinensis extract ethyl acetate fraction 2g

1g lactose

The above ingredients were mixed and filled in airtight cloth to prepare a powder.

2. Preparation of Tablets

Schizandra chinensis extract ethyl acetate fraction 100mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

3. Preparation of Capsule

Schizandra chinensis extract ethyl acetate fraction 100mg

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, the capsule was prepared by filling in gelatin capsules according to the conventional method for producing a capsule.

The compositions of the present invention may be added to foods or beverages for the purpose of ameliorating diseases caused by arthritis. In this case, the amount added may be added in an amount of 1 to 5% by weight, preferably 3% by weight of the total food weight in the case of the health food composition, and in the case of the health beverage composition, 1 to 3g, preferably 2g Can be added in proportion.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, etc. as additional components, as in the general beverage. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as tautin and stevia extract, synthetic sweeteners such as saccharin, aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 2-5 g, preferably about 4 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonic acid. Carbonating agents and the like used in beverages. In addition, the composition of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not critical but is usually selected in the range of 1 to 5 parts by weight per 100 parts by weight of the composition of the present invention.

Formulation Example 2 : Manufacture of food

Foods containing the Schisandra chinensis extract of the present invention were prepared as follows.

1. Preparation of Cooking Seasonings

Schisandra chinensis extract of the present invention was prepared in 20 to 95% by weight for health promotion cooking seasoning.

2. Preparation of Tomato Ketchup and Sauce

0.2 ~ 1.0% by weight of Schizandra extract of the present invention was added to tomato ketchup or sauce to prepare tomato ketchup or sauce for health promotion.

3. Manufacturing of Flour Foods

0.5 ~ 5.0% by weight of Schizandra chinensis extract of the present invention was added to the flour, and using the mixture to prepare bread, cakes, cookies, crackers and noodles to prepare foods for health promotion.

4. Preparation of soups and gravy

0.1-5.0 wt% of Schizandra chinensis extract of the present invention was added to soups and broths to prepare meat products for health promotion, soups and noodles for noodles.

5. Preparation of Ground Beef

10 wt% of Schisandra chinensis extract of the present invention was added to ground beef to prepare a ground beef for health promotion.

6. Manufacture of Dairy Products

5 to 10% by weight of Schizandra chinensis extract of the present invention was added to milk, and the milk was used to prepare various dairy products such as butter and ice cream.

7. Manufacture of wire

Brown rice, barley, glutinous rice, and yulmu were alphad by a known method, and then dried and roasted to prepare a powder having a particle size of 60 mesh using a grinder.

Black beans, black sesame seeds, and perilla were also steamed and dried by a known method, and then ground to a powder having a particle size of 60 mesh.

The Schizandra chinensis extract of the present invention was decompressed and concentrated in a vacuum concentrator, and the dried product obtained by drying with a sprayer or a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

It was prepared by combining the dry powder of the grains, seeds and Schisandra chinensis extract prepared in the following ratio.

Cereals (30% by weight brown rice, 15% by weight radish, 20% by weight barley),

Seeds (7% by weight perilla, 8% by weight black beans, 7% by weight black sesame),

Dry powder of Schizandra chinensis extract (3% by weight),

Ganoderma lucidum (0.5% by weight),

Foxglove (0.5 wt%)

Formulation Example 3 : Preparation of Beverages

1. Preparation of Carbonated Drinks

5-10% of sugar, 0.05-0.3% citric acid, 0.005-0.02% caramel, 0.1-1% of vitamin C are mixed, and 79-94% purified water is mixed to make syrup, and the syrup is 85-98 Sterilizing at 20 ℃ for 180 seconds to mix with a cooling water 1: 1: ratio of carbon dioxide gas was injected 0.5 ~ 0.82% to prepare a carbonated beverage containing Schizandra extract of the present invention.

2. Manufacture of health drinks

Instant sterilization by homogeneously mixing minor ingredients such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%) and Schizandra chinensis extract Health drinks were prepared by packaging in small packaging containers such as plastic bottles.

3. Preparation of Vegetable Juice

5 g of Schisandra chinensis extract of the present invention was added to 1,000 ml of tomato or carrot juice to prepare a vegetable juice for health promotion.

4. Preparation of Fruit Juice

1 g of Schizandra chinensis extract of the present invention was added to 1,000 ml of apple or grape juice to prepare fruit juice for health promotion.

Hereinafter, preferred examples are provided to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

Example 1 : Preparation of Schizandra chinensis Extract

1.Schisandra chinensis extract

1) Schizandra Water Extract

After cutting 3 kg of dried Schisandra chinensis fruits, stirring and extracting was repeated five times for 24 hours each with water, and then filtered through a filter paper, and the filtrate was concentrated under reduced pressure to obtain a water extract.

2) Schizandra Organic Solvent Extract

After cutting 3 kg of dried Schisandra chinensis fruit, the mixture was extracted by stirring repeatedly for 5 hours with methanol for 24 hours, filtered through a filter paper, and the filtrate was concentrated under reduced pressure to obtain a methanol extract.

2. Isolation of Schizandra chinensis Extract

Distilled water was added to the methanol extract obtained in 2) and extracted with hexane. The extracted residue was reextracted with chloroform, and the extracted residue was extracted with ethyl acetate again.

The ethyl acetate extract was checked for the number of active substances using TLC (Thin Layer Chromatography), and the active fractions were purified using silica, C ₁₈ and NH ₂ chromatography columns.

Extraction, separation and purification of the Schizandra chinensis extract of the present invention is shown in FIG.

Experimental Example 1 : Determination of Hyaluronidase Activity of Schizandra chinensis Extract

In order to determine the inhibitory activity of hyaluronides of the Schizandra chinensis extract of the present invention, the activity was measured using the Morgan-Elson Assay method.

Hyaluronic acid dissolved in phosphate buffer solution and Schizandra chinensis extract dissolved in DMSO, respectively, were added and incubated at 37 ° C. for 20 minutes. After incubation, the compound 48/80 solution was added and then secondary cultured. After hyaluronic acid was added and tertiary incubation, K ₃ BO ₃ was added thereto, and the reaction was stopped by heating in a boiling water. After centrifugation, N-acetylglucosamine (N-acetylglucosamine) produced by the decomposition of hyaluronic acid by hyaluronates was verified by colorimetric determination at 585 nm using DMAB (p-dimetylaminobenzaldehyde).

The results are shown in Table 1.

Inhibition of Hyaluronidase Activity of Schisandra chinensis Fractions Methanol extract n-hexanesoluble fraction Chloroform Soluble Fraction Ethyl Acetate Soluble Fraction % Activity inhibition 62.6 ± 0.71 10.0 ± 0.28 2.6 ± 0.07 59.0 ± 2.79

As shown in Table 1, Schizandra extract of the present invention can be seen that significantly inhibit the activity of hyaluronidase.

Experimental Example 2 : Effect of Schizandra chinensis Extract on the Growth of Articular Chondrocytes (C28 / I2 Cells) and Extracellular Matrix Synthesis

In order to investigate the effects of the hyaluronatease enzyme inhibitory activity and the extracellular matrix synthesis of chondrocytes on the human costal chondrocytes of Schizandra chinensis extract, the expression pattern of collagen type II, which is a specific extracellular matrix of cartilage, was determined by RT. -Confirmed by PCR.

Human costal chondrocytes were cultured in 1) control group (chondrocytes), 2) hyaluronidase treated group, 3) Schizandra extract and hyaluronidase treated groups, and observed for 24 ~ 72 hours.

After removing monolayer cultured chondrocytes, total RNA was isolated using Trizol Reagent (RNA extraction Kit, GIBCOBRL). Complementary DNA (cDNA) was prepared using AMV reverse transcriptase (BMS) for the same amount of RNA for each group, which was then amplified by PCR using primers for collagen type II and GAPDH. After the change was confirmed by electrophoresis on the agarose gel.

The results are shown in FIG.

As shown in Figure 2, the treatment of the Schizandra chinensis extract of the present invention can be seen that the collagen expression is increased than the hyaluronidase alone treatment group, which is that Schizandra extract inhibits the synthesis of extracellular matrix of chondrocytes. It can be seen that it suppresses again.

Therefore, it can be seen that the Schizandra chinensis extract of the present invention inhibits the activity of hyaluronides.

The composition containing the Schizandra chinensis extract of the present invention as an active ingredient is excellent in inhibiting hyaluronic acid degradation by hyaluronic acid in the joints commonly occurring in osteoarthritis patients, and hyaluronicase inhibits the extracellular matrix synthesis of chondrocytes. It can be usefully used in medicine and health foods for the prevention and treatment of arthritis by suppressing that.

Claims (7)

Arthritis prevention and treatment composition containing Schizandra chinensis extract as an active ingredient. According to claim 1, Schisandra chinensis extract is a composition for preventing and treating arthritis, characterized in that the extraction with water or an organic solvent. The method of claim 2, wherein the organic solvent is a composition for preventing and treating arthritis, characterized in that selected from alcohol, chloroform, hexane, ethyl acetate. The extract of Schizandra chinensis extract according to claim 1, wherein Schizandra chinensis extract is extracted with methanol, distilled water is added to the methanol extract, and then extracted with hexane. Arthritis prevention and treatment composition, characterized in that. A pharmaceutical composition for the prevention and treatment of arthritis, comprising the composition of any one of claims 1 to 4. Food composition for the prevention and treatment of arthritis, containing the composition of any one of claims 1 to 4. Hyaluronidase inhibitor containing Schizandra chinensis extract as an active ingredient.