KR20230044120A - Composition for prevention, amelioration or treatment of arthritis comprising Glycine soja extract as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating or treating arthritis containing a Glycine soja extract as an active component. The Glycine soja extract, specifically, a Glycine soja leaf or stem extract of the present invention, compared to an extract of Glycine soja fruits, reduces the content of prostaglandin E2 (PGE2), an inflammatory mediator, has an excellent effect of inhibiting MMP-13, a cartilage matrix decomposition enzyme, exhibits an analgesic effect when treated with Glycine soja leaf or stem extract in animal models suffering from pain, and thus has an effect of relieving edema. Therefore, the composition of the present invention can be usefully used for preventing, alleviating or treating arthritis.

Description

Composition for prevention, amelioration or treatment of arthritis comprising Glycine soja extract as effective component}

The present invention relates to a composition for the prevention, improvement or treatment of arthritis containing a stone soybean extract as an active ingredient.

As we enter an aging society, inflammatory diseases including arthritis are socially emerging regardless of gender. Inflammatory diseases caused by this inflammatory response include gastritis, colitis, arthritis, nephritis, hepatitis, arteriosclerosis, cancer, or degenerative diseases. Among them, effective drugs or treatments for the prevention and treatment of arthritis diseases have not been developed so far. Arthritis refers to an inflammatory change in a joint due to various causes such as aging, mechanical damage, and immune abnormality.

Among the above-mentioned arthritis, osteoarthritis (osteoarthritis) is a kind of arthritis, also called degenerative arthritis, refers to arthritis caused by degenerative changes in cartilage and surrounding bones in synovial joints. That is, osteoarthritis is a disease characterized by gradual loss of articular cartilage, hypertrophy of bones located below the cartilage, bone formation at the joint edge, and non-specific synovial inflammation. Osteoarthritis is a disease caused by damage to cartilage due to aging or excessive physical pressure (eg, obesity, trauma, etc.). Therefore, osteoarthritis shows severe pain and movement disorders in joints that receive a lot of weight, that is, knee (knee) joints, hip (hip) joints, etc., and even causes deformation of the joints when left unattended for a long period of time.

On the other hand, stone beans ( Glycine soja ) is an annual plant belonging to the legume family (Zingiberaceae) and is 2m long and has brown coarse hairs throughout. The leaves are alternate, the petioles are long, and the leaves are 3. Fruits are 2-3 cm long, hairy and similar to bean pods, and seeds are oval or kidney-like and slightly flat.

Dolbean is considered to be the origin of soybean ( Glycine max ), and is currently recognized as edible, but is rarely used for actual food. Most of them grow naturally in nature and have strong genes, so they are sometimes cultivated for research on genetic improvement of soybeans.

As prior art related to arthritis, Korean Patent Publication No. 2007-0109004 discloses a pharmaceutical composition for the prevention and treatment of bone metabolic diseases, diseases caused by oxidative stress, and inflammatory diseases, including herbal medicine extracts, and Korean Patent Publication No. 2005-0109004. No. 0019830 discloses a functional food for preventing autoimmune diseases containing an extract of herbal medicine as an active ingredient, but a composition for preventing, improving or treating arthritis containing the extract of dolbean as an active ingredient of the present invention has not been disclosed.

The present invention was derived from the above needs, and the present invention provides a composition for preventing, improving or treating arthritis containing a stone bean extract as an active ingredient, and a stone bean extract of the present invention, specifically a stone bean leaf or stem extract. Compared to this stone bean fruit extract, it reduces the content of prostaglandin E2 (PGE2), an inflammatory mediator, and has an excellent inhibitory effect on MMP-13, a cartilage matrix decomposing enzyme. The present invention was completed by confirming that there is an effect of showing the effect and alleviating edema.

In order to achieve the above object, the present invention provides a health functional food composition for preventing or improving arthritis containing a stone soybean extract as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of arthritis containing a stone soybean extract as an active ingredient.

In addition, the present invention provides a feed additive for preventing or improving arthritis containing a stone soybean extract as an active ingredient.

In addition, the present invention provides a veterinary composition for the prevention or treatment of arthritis in animals other than humans, containing a stone soybean extract as an active ingredient.

The present invention relates to a composition for the prevention, improvement or treatment of arthritis containing an extract of a soybean as an active ingredient, wherein the extract of a soybean, specifically a soybean leaf or stem extract, is an inflammatory mediator, prostaglandin E2 (PGE2 ) content, and has an excellent effect of inhibiting MMP-13, a cartilage matrix decomposition enzyme, and exhibits analgesic effect and relieves edema when treating pain-induced animal models with leaf or stem extracts. The composition of the present invention can be usefully used for preventing, improving or treating arthritis.

Figure 1 is the result of confirming the change in PGE2 content according to the treatment of the extract for each part of the soybean of the present invention. #### indicates that the PGE2 content of the IL-1β treated group was significantly increased compared to the untreated control group (Con), p<0.0001. *, **, *** means that the PGE2 content of the extract treatment group for each part of the soybean of the present invention was significantly decreased compared to the IL-1β treatment group, * is p <0.05, ** is p <0.01, *** indicates p<0.001.
Figure 2 is the result of confirming the change in the content of MMP-13 according to the treatment of the extract for each part of the soybean of the present invention. #### indicates that the MMP-13 content of the IL-1β treated group was significantly increased compared to the untreated control group (Con), p<0.0001. *, **, **** indicate that the MMP-13 content of the extract treatment group for each part of the soybean of the present invention was significantly decreased compared to the IL-1β treatment group, * is p <0.05, and ** is p < 0.01, and **** is p<0.0001.
Figure 3 is a result of confirming the analgesic effect on pain according to the administration of the leaf or stem extract in an animal model in which pain was induced by acetic acid. Diclofenac is a positive control. * indicates a statistically significant decrease in the number of turns of the body for 10 minutes in the sample-treated group compared to the control group (con) not treated with the sample after induction of pain, p<0.05.
Figure 4 is the result of confirming the foot edema relief effect according to the administration of stone bean leaf or stem extract in an animal model induced inflammation with carrageenan. ** indicates that the foot edema of the dolko leaf extract-treated group was significantly reduced compared to the inflammation-induced control group (con), p<0.01.

In order to achieve the object of the present invention, the present invention provides a health functional food composition for preventing or improving arthritis containing an extract of dolkong ( Glycine soja ) as an active ingredient.

The stone bean can be used any part of the stone bean plant, but preferably the stone bean leaf or stem part is used, more preferably the stone bean leaf is used, but is not limited thereto.

The arthritis is preferably, but not limited to, osteoarthritis, and the composition is characterized in that it inhibits inflammatory cytokines.

The stone soybean extract may be prepared by a method comprising the following steps, but is not limited thereto:

1) extracting by adding an extraction solvent to the leaves or stems of soybeans;

2) filtering the extract of step 1); and

3) preparing an extract by concentrating the filtered extract of step 2) under reduced pressure and drying it.

In step 1), the extraction solvent is preferably water, C ₁ ~ C ₄ lower alcohol or a mixture thereof, more preferably 70% (v / v) ethanol extract, but is not limited thereto.

In the above production method, all conventional methods known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction, can be used for extraction of the soybean leaves or stems. The vacuum concentration in step 3) is preferably performed using a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.

The composition is preferably prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup and beverage, but is not limited thereto.

When using the health functional food composition of the present invention as a food additive, the stone soybean extract may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to conventional methods. The mixing amount of the active ingredient can be suitably determined according to its purpose of use (prevention, health or therapeutic treatment). In general, when preparing food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. There is no particular limitation on the type of food. Examples of foods to which the extract or its fraction can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea , drinks, alcoholic beverages, and vitamin complexes, and includes all health functional foods in the conventional sense.

When the composition of the present invention is used as a health drink, it may contain various flavoring agents or natural carbohydrates as additional components, like conventional drinks. The aforementioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclotenstrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention. The composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal agents, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonic acid It may contain a carbonation agent used in beverages and the like. In addition, the composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The ratio of these additives is not very important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention relates to a pharmaceutical composition for the prevention or treatment of arthritis containing a stone soybean extract as an active ingredient.

The arthritis is preferably osteoarthritis, but is not limited thereto.

The composition of the present invention is preferably prepared in any one formulation selected from capsules, powders, granules, tablets, suspensions, emulsions, syrups and aerosols, but is not limited thereto.

The composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the above active ingredient, and may be in various oral or parenteral formulations. When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include capsules, powders, granules, tablets, pills, and the like, and these solid preparations include at least one excipient in one or more compounds, for example, starch, calcium carbonate, sucrose or lactose ( lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, emulsions, syrups, aerosols, etc., and various excipients such as wetting agents, sweeteners, aromatics, and preservatives may be included in addition to water and liquid paraffin, which are commonly used simple diluents. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried formulations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending solvents. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used. In the case of parenteral administration, it is preferable to select an external skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine intrathecal or intracerebrovascular injection method.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the level of the effective amount depends on the type, severity, and activity of the drug of the patient's disease. , sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered single or multiple times. Considering all of the above factors, it is important to administer an amount that can obtain the maximum effect with the minimum amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the composition of the present invention varies in its range depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate and severity of the disease, and the daily dose is based on the amount of dolbean extract 0.01 to 2,000 mg/kg, preferably 30 to 500 mg/kg, more preferably 50 to 300 mg/kg, and may be administered 1 to 6 times a day. 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.

In addition, the present invention provides a feed additive for preventing or improving arthritis containing a stone soybean extract as an active ingredient.

The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. In the present invention, the term 'feed' may refer to any natural or artificial diet, one meal, etc., or a component of the one meal meal, suitable for or suitable for consumption by animals. The type of feed is not particularly limited, and feeds commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products; Animal feed such as proteins, inorganic materials, oils, mineral oils, oils, single cell proteins, zooplankton, or food may be mentioned. These may be used alone or in combination of two or more.

In addition, the present invention provides a veterinary composition for the prevention or treatment of arthritis containing a stone soybean extract as an active ingredient.

The veterinary composition containing the extract of dol soybean of the present invention may further include appropriate excipients and diluents according to conventional methods. Examples of excipients and diluents that may be included in the veterinary composition containing the stone soybean extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium Phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostearate, polysorbate 60, methylparaben, propylparaben and mineral oil.

The veterinary composition containing the soybean extract according to the present invention may further include a filler, an anti-coagulant, a lubricant, a wetting agent, a spice, an emulsifier, a preservative, etc. The veterinary composition according to the present invention is active after administration to an animal It can be formulated using methods well known in the art to provide rapid, sustained or delayed release of the ingredients, and the dosage form can be powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft Alternatively, it may be in the form of a hard gelatin capsule, a suppository, a sterile solution for injection, or a sterile external preparation.

The veterinary composition according to the present invention may include the extract in an effective amount or as an active ingredient depending on its use. The effective amount can be appropriately selected according to the individual animal. The severity of the disease or condition, the subject's age, weight, health, sex, the subject's sensitivity to the extract, the administration time, administration route and excretion rate, administration period, factors including other compositions combined or used simultaneously with the composition, and other It may be determined according to factors well known in the physiological or veterinary field.

Hereinafter, the present invention will be described in more detail using examples. These examples are only for explaining the present invention in more detail, and it is obvious to those skilled in the art that the scope of the present invention is not limited thereto.

Example 1. Preparation of extracts for each part of soybean

For each of 1kg of dolbean fruit, leaf or stem, 15 ℓ of 70% (v / v) ethanol was added, extracted at 85 ° C. for 3 hours, and the filtered solution was concentrated under reduced pressure at 45 ° C. DC), soybean leaf extract (DC-L) and soybean stem extract (DC-S) were obtained, respectively.

Example 2. Evaluation of anti-inflammatory efficacy in chondrocytes

In human chondrocytes (SW1353), changes in the content of prostaglandin E2 (PGE2), an inflammatory mediator, according to the treatment of the extract were confirmed. In addition, it was confirmed whether MMP-13, a cartilage matrix degrading enzyme activated by an increase in inflammatory cytokines, was inhibited.

[Cell culture]

After culturing human chondrocytes (SW1353) using a culture medium in which 10% FBS was added to DMEM/F12 (Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12), 50 to 200 μg/ml of extract for each part of the eagle soybean was added. After each treatment and incubation for 2 hours, IL-1β (10 μg/ml) was treated for 24 hours.

(1) Confirmation of inhibitory effect on PGE2 production induced by IL-1β

In order to evaluate the anti-inflammatory effect of each part of the soybean extract, the inhibitory effect on the production of PGE2, an inflammatory mediator, was measured using an ELISA kit (R&D Systems).

As a result, the production of PGE2 was significantly increased in chondrocytes by IL-1β treatment, and in the cells pre-treated with the extract of each part of the soybean, the production of PGE2 was significantly reduced at a concentration of 100 μg/ml. In the treated cells, PGE2 production was significantly reduced even at a concentration of 50 μg/ml (FIG. 1).

(2) Confirmation of IL-1β-mediated reduction of MMP-13

The activation and synthesis of MMPs following the secretion of inflammatory cytokines degrades cartilage matrix molecules (ECM) constituting cartilage tissue. It was confirmed using

As a result, the expression level of MMP-13 increased in the culture supernatant treated with IL-1β for 24 hours, but the expression level of MMP-13 was significantly decreased in the cells pre-treated with the leaf extract or the stem extract of the stone bean (Fig. 2). .

Example 3. Evaluation of analgesic efficacy in pain-induced animal models by acetic acid

7-week-old ICR mice were acclimatized for one week, fasted for 18 hours or longer, and then orally administered with dolbean leaf or stem extract (300 mg/kg) and positive control diclofenac (10 mg/kg). One hour after administration, 0.75% (v/v) acetic acid (acetic acid) was intraperitoneally administered at a dose of 10 ml/kg, and the number of times the mouse twisted its body was observed for 10 minutes after administration.

As a result, intraperitoneal administration of acetic acid caused capillary damage and pain, resulting in an increase in the number of twists of the mouse's body, and when the extract of the leaf or stem of the present invention was administered, the body of the mouse was significantly It was confirmed that the number of beats decreased (FIG. 3).

Example 4. Evaluation of efficacy of relieving foot edema in an animal model inducing inflammation by carrageenan

After 7-week-old ICR mice were acclimatized for one week, they were orally administered with stone bean leaf or stem extract (300mg/kg) and indomethacin (5mg/kg) as a positive control, and 1% (w/v) carrageenan 1 hour after administration. 50 ul was administered to the sole of the right hind limb, and 5 hours later, the size of the edema of the foot of the right hind limb was compared using a edema measuring device (Plethysmometer, Bioseb, USA).

As a result, in the carrageenan-induced foot edema animal model, it was confirmed that the foot edema caused by the extract administration was significantly reduced at 5 hours after the edema was induced (FIG. 4).

Claims (10)

Stone beans ( Glycine soja ) Health functional food composition for preventing or improving arthritis containing an extract as an active ingredient. The health functional food composition for preventing or improving arthritis according to claim 1, wherein the stone bean is a leaf or stem portion of the stone bean. The functional food composition for preventing or improving arthritis according to claim 1, wherein the arthritis is osteoarthritis. The functional food composition for preventing or improving arthritis according to claim 1, wherein the composition inhibits inflammatory cytokines. The health functional food composition for preventing or improving arthritis according to claim 1, wherein the extraction solvent of the soybean extract is water, C ₁ ~ C ₄ lower alcohol, or a mixture thereof. The functional food composition for preventing or improving arthritis according to claim 1, wherein the composition is prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup and beverage. A pharmaceutical composition for the prevention or treatment of arthritis containing a stone bean extract as an active ingredient. The pharmaceutical composition for preventing or treating arthritis according to claim 7, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient. A feed additive for preventing or improving arthritis containing a stone bean extract as an active ingredient. A veterinary composition for the prevention or treatment of arthritis containing a stone bean extract as an active ingredient.

Images