WO2019177327A1 - Composition comprising schisandra chinensis extract as effective ingredient for prevention, alleviation, or treatment of arthritis - Google Patents

Abstract

The present invention relates to a composition comprising an extract from Schisandra chinensis leaves or stems as an effective ingredient for prevention, alleviation, or treatment of arthritis. Specifically, the extract from Schisandra chinensis leaves or stems according to the present invention is not cytotoxic, has an effect of inhibiting inflammation-inducted NO production, reduces a thickness of a foot swollen by gout arthritis, and decreases a content of IL-1β. The extract from Schisandra chinensis leaves according to the present invention has an effect of increasing weight bearing reduced in an osteoarthritis animal model with a statistical significance. Thus, the extract can be usefully availed for prevention, alleviation, or treatment of arthritis.

Description

Composition for the prevention, improvement or treatment of arthritis containing Schizandra chinensis extract as an active ingredient

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

As we move into an aging society, inflammatory diseases, including arthritis, are emerging socially regardless of gender. Inflammatory diseases caused by these inflammatory reactions include gastritis, colitis, arthritis, nephritis, hepatitis, arteriosclerosis, cancer or degenerative diseases. To date, effective drugs or treatments for the prevention and treatment of arthritis diseases have not been developed. Arthritis refers to the development of inflammatory changes in the joints due to various causes such as aging, mechanical damage, and immune disorders.

Of the above arthritis, osteoarthritis is a type of arthritis, also called degenerative arthritis, and refers to arthritis caused by degenerative changes in cartilage and surrounding bone in lubricated joints. In other words, osteoarthritis is a disease characterized by gradual loss of articular cartilage, hypertrophy of bone located below the cartilage, bone formation at the edge of the joint, and nonspecific synovial inflammation. Osteoarthritis is a disease caused by cartilage damage caused by aging or excessive physical pressure (eg, obesity, trauma, etc.). Therefore, osteoarthritis represents severe pain and movement disorders such as knee joints, knee joints, hip joints, etc., which are heavily weighted, resulting in deformation of joints when left for a long time.

In addition, gouty arthritis is an inflammation caused by the deposition of uric acid in the spaces and tissues of the joints. Urate is increased by increasing the concentration of uric acid (the product of the body's metabolism of purine, which is obtained through food). (Uric acid is in the form of urate in the blood, body fluids, and joint fluids.) Crystals deposit in the cartilage, tendons, and surrounding tissue of the joint. This phenomenon causes inflammation of the joints, leading to recurrent attacks with extreme pain.

On the other hand, Schizandra chinensis ) is a dicotyledonous plant, a deciduous vine of Magnolia schizandra. Mainly growing in the valley, the trunk is brown and has the property of climbing trees. The leaves are alternate, wide oval, long oval or egg-shaped, with hairs on the rear leaf veins and tooth-shaped teeth on the edges. Flowers bloom in June-July, single flowers, slightly reddish yellowish white. After flowering, the female flower jaw grows 3 ~ 5cm long and the fruit runs with water. Fruits are berry (漿果), almost round, with several spikes. It ripens red in August-September and contains 1-2 reddish brown seeds. Young sprouts eat as herbs. The fruit is mixed with five flavors of sour, sweet, bitter, salty and spicy. It is prescribed for nourishment, tonic, Jinhae, expectoration, and cold in Chinese medicine. It is prescribed for seawater, oil well, gugal, dohan, acute hepatitis. . In the private sector, Schisandra tea is made and drunk.

The prior art related to Schisandra chinensis No. 2007-0109004 discloses a pharmaceutical composition for the prevention and treatment of bone metabolism disease, oxidative stress, inflammatory diseases, including Schisandra chinensis extract, Korea Patent Publication No. 2005-09 Although 0019830 discloses a functional food for preventing autoimmune diseases containing Schizandra chinensis extract as an active ingredient, a composition for preventing, improving or treating arthritis containing Schizandra extract of the present invention as an active ingredient is not disclosed.

The present invention is derived from the above demands, Schisandra ( Schisandra) chinensis) provides a composition for the prevention, improvement or treatment of arthritis containing leaf or stem extract as an active ingredient, Schisandra of the present invention ( Schisandra chinensi s) leaf or stem extract is not cytotoxic, has the effect of inhibiting the production of inflammation-induced NO, and statistically significantly reduces the increased foot thickness caused by gouty arthritis, as well as the content of IL-1β The effect of reducing was superior to the Schisandra chinensis fruits, and when the Schisandra chinensis extract was administered in an osteoarthritis animal model, the present invention was completed by confirming that the weight loss rate was statistically significantly increased.

In order to achieve the above object, the present invention Schizandra ( Schisandra) chinensi s) It provides a health functional food composition for the prevention or improvement of arthritis containing leaf or stem extract as an active ingredient.

The present invention also provides a pharmaceutical composition for the prevention or treatment of arthritis, containing Schizandra leaf or stem extract as an active ingredient.

The present invention Schisandra ( Schisandra) chinensi s) relates to a composition for the prevention, improvement or treatment of arthritis containing a leaf or stem extract as an active ingredient, specifically, Schisandra of the present invention ( Schisandra chinensi s) leaf or stem extract is not cytotoxic, has the effect of inhibiting the production of NO induced by inflammation, and statistically significantly reduces the thickness of the feet increased by gouty arthritis, as well as the content of IL-1β In the osteoarthritis animal model, administration of the Schisandra chinensis leaf extract of the present invention has a statistically significant effect of reducing the weight-bearing rate.

1 is a result confirming the effect of the Schizandra fruit, leaf or stem extract of the present invention on the survival rate of RAW 264.7 cells. NMMA is a positive control group, N-Monomethyl arginine.

Figure 2 shows that the production of NO increased by the treatment of LPS (lipopolysaccharide) in RAW 264.7 cells, the result of confirming that the production of NO increased by LPS by treating the Schizandra leaf or stem extract of the present invention. Con is LPS-treated RAW264,7 cells, *, **** is a statistically significant NO production reduction compared to Con, * is p <0.05, **** is p <0.0001.

Figure 3 is a result confirming the effect of reducing the foot thickness (foot thickness) in C57BL6 mice induced gouty arthritis by MSU (monosodium ureate). Con is a control group that did not induce gouty arthritis, MSU is a group that caused gouty arthritis by administration of MSU, Schisandra chinensis-Schisandra chinensis, Schisandra chinensis-Schisandra chinensis, MSU + Schisandra chinensis fruit, leaf or stem extract Each group was treated and COL was colchicine, a positive control group. #### means that the foot thickness of the gouty arthritis group (MSU) by MSU induction was significantly increased compared to the normal group (Con), which means p <0.0001, and *, **** Compared with the gouty arthritis group, Schisandra chinensis leaf or stem extract treatment group showed statistically significant decrease in foot thickness. * Is p <0.05 and **** is p <0.0001.

Figure 4 is a result confirming the reducing effect of IL-1β in C57BL6 mice induced gouty arthritis by MSU (monosodium ureate). Con is a control group that did not induce gouty arthritis, MSU is a group that caused gouty arthritis by administration of MSU, Schisandra chinensis-Schisandra chinensis, Schisandra chinensis-Schisandra chinensis, MSU + Schisandra chinensis fruit, leaf or stem extract Each group was treated and COL was colchicine, a positive control group. #### indicates that the foot thickness of the gout arthritis group induced by MSU was significantly increased compared to the normal group (Con), which means p <0.0001, and * and *** are gout arthritis groups induced by MSU. Compared with (MSU), Schizandra chinensis extract treatment group and positive control group colchicine (COL) showed statistically significant reduction in foot thickness. * Is p <0.05 and *** is p <0.001.

Figure 5 is the result showing the hind limb weight load rate (%) of the SD rat. Con is a negative control group, MIA (monosodium iodoacetate) is a osteoarthritis-inducing group, Indomethacin is a positive control group, Schisandra chinensis leaf-300 is a group of 300 mg / ㎏ MIA and Schisandra chinensis extract, Schisandra chinensis -150 MIA and Schisandra chinensis Fructus extract 150 mg / kg. #### means that the weight-bearing rate of the osteoarthritis-induced group by MIA was significantly reduced compared to the negative control group, which means p <0.0001. *, **, ***, **** is an experimental group of MIA and Schisandra chinensis extract (300mg / kg, 150mg / kg) in parallel treatment; Alternatively, the weight-bearing rate (%) of the positive control group treated with MIA and indomethacin increased compared to the osteoarthritis-induced group treated with MIA, where * is p <0.05, ** is p <0.01, and *** Is p <0.001 and **** means p <0.0001.

The present invention Schisandra ( Schisandra) chinensi s) relates to a nutraceutical composition for the prevention or improvement of arthritis containing leaf or stem extract as an active ingredient.

Preferably, the arthritis is osteoarthritis or gouty arthritis, but is not limited thereto. The composition may be characterized by inhibiting inflammatory cytokines and pain, and the pain may be pain due to arthritis.

The Schisandra chinensis leaf or stem 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 Schizandra leaves or stems;

2) filtering the extract of step 1); And

3) concentration of the filtered extract of step 2) under reduced pressure and drying to prepare an extract.

The extraction solvent in step 1) is preferably water, lower alcohols of C ₁ ~ C ₄ or mixtures thereof, more preferably 70% (v / v) ethanol extract, but is not limited thereto.

In the above production method, the extraction of Schisandra chinensis leaves or stems may use all conventional methods known in the art, such as filtration, hot water extraction, dipping extraction, reflux cooling extraction, and ultrasonic extraction. Concentration under reduced pressure of step 3) is preferably a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.

The composition is preferably, but not limited to, prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrups and beverages.

When the health functional food composition of the present invention is used as a food additive, the Schizandra leaf or stem extract may be added as it is, or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method. The blending amount of the active ingredient can be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). Generally, in the preparation of food or beverages, the composition of the present invention is added in an amount of up to 15 parts by weight, preferably up to 10 parts by weight based on the raw materials. However, in the case of long-term intake for health and hygiene or health control purposes, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. There is no particular limitation on the kind of food. Examples of foods to which the extract or fractions thereof may be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, drinks, tea , Drinks, alcoholic beverages and vitamin complexes, and includes all the health functional foods in the conventional sense.

When the composition of the present invention is used as a health beverage, various flavors, natural carbohydrates, and the like may be contained as additional components, as in general beverages. The above-mentioned natural carbohydrates are sugars such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as textine and cyclotenstrin, xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio 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. In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols. And carbonation agents used in carbonated beverages. In addition, the composition of the present invention may contain a pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical, but the composition of the present invention is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.

The present invention also relates to a pharmaceutical composition for the prevention or treatment of arthritis, containing Schizandra leaf or stem extract as an active ingredient. The arthritis is preferably, but not limited to, osteoarthritis or gouty arthritis.

The composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient, and may be various oral or parenteral formulations. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include capsules, powders, granules, tablets, pills, and the like, which may comprise at least one excipient such as starch, calcium carbonate, sucrose or lactose (at least one compound). lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, emulsions, syrups, aerosols and the like, and may include various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc., in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycero gelatin and the like can be used. For parenteral administration, it is desirable to select either external or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dura or cerebrovascular injections.

The pharmaceutical composition according to the invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, the effective amount of the level of the disease, the severity, the drug activity of the patient , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of the drug, and other factors well known in the medical arts. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with a minimum amount without side effects, which can be readily determined by one skilled in the art.

The dosage of the composition of the present invention varies depending on the weight, age, sex, health condition, diet, time of administration, administration method, excretion rate and severity of the disease of the patient, the daily dosage of Schisandra chinensis leaf or stem extract 0.01-2,000 mg / kg, preferably 30-500 mg / kg, more preferably 50-300 mg / kg, based on the amount, can be administered 1-6 times per day. The compositions of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for explaining the present invention in more detail, it is obvious to those skilled in the art that the scope of the present invention is not limited by them.

Example 1 Preparation of Schisandra chinensis Leaves, Stems and Fruit Extracts

15 L of 70% (v / v) ethanol was added to each of 1 kg of Schizandra chinensis leaves, 1 kg Schizandra chinensis or 1 kg Schizandra chinensis, and extracted at 85 DEG C for 3 hours, and then the filtrate was filtered at 45 DEG C. Concentration under reduced pressure and drying yielded Schizandra leaf extract, Schizandra stem extract or Schizandra fruit extract.

Example 2 Evaluation of Anti-inflammatory Effect of Schizandra chinensis Extract in RAW 264.7 Cells

1) MTT Assay

RAW 264.7 cells, which are mouse macrophage lines, were dispensed in 96-well plates, followed by incubation for 24 hours or 48 hours by treatment of Schizandra ethanol extract prepared in Example 1 by concentration (25, 50 ㎍ / ㎖) It was. After culturing the cells, the MTT solution was added and reacted for 4 hours, and then the absorbance of 540 nm was measured using an ELISA reader. The relative cell viability was calculated by comparison with the control group.

As a result, as shown in Figure 1, Schizandra ethanol extract was confirmed that there is no cytotoxicity does not affect the cell viability.

2) NO measurement

LPS (500 ng / ml) was treated to RAW 264.7 cells divided into 48-well plates at an appropriate concentration, and the Schisandra chinensis ethanol extract prepared in Example 1 was treated by concentration (25, 50 µg / ml) for 24 hours. Incubated. After taking the culture supernatant, the amount of NO was measured according to the manufacturer's method using Griess reagent.

As a result, as shown in FIG. 2, the amount of NO production increased by LPS was reduced in a concentration-dependent manner by the treatment of Schisandra chinensis ethanol extract, and in particular, the reducing effect of Schisandra chinensis leaf or stem extract was remarkable.

Example  3. To MSU (Monosodium ureate)  Evaluation of Efficacy of Schizandra chinensis Extract in Induced Gouty Arthritis Animal Model

 In order to confirm the efficacy of the extract from Schizandra chinensis extract in Example 1 in MSU (Monosodium ureate) -induced gouty arthritis disease animal model, the effect of reducing the inflammatory cytokine IL-1β was confirmed.

70% (v / v) ethanol extract of Schizandra chinensis was administered orally to 7-week-old C57BL6 mice at a concentration of 300 mg / kg. One hour after administration of the Schizandra chinensis ethanol extract, 4 mg of MSU was suspended in 50 µl of PBS containing 2.5% Tween 80 and injected into the right paw tissue to induce gouty arthritis. The drug was administered once a day for 4 days, followed by necropsy on the 5th day of MSU induction.

Subsequently, the paw tissue obtained by the autopsy was immersed in RIPA buffer solution, and the tissue was crushed to measure the expression level of IL-1β protein, which is an inflammatory marker in the supernatant, by ELISA, and cholchicine as a positive control. Was used.

As a result, as shown in FIG. 3, it was confirmed that the thickness of the feet in the Schisandra chinensis leaf extract and Schisandra chinensis extract was statistically significantly reduced, and as shown in FIG. 4, IL-1β increased due to gouty arthritis of Schisandra chinensis. It was confirmed that significantly decreased by administration of the extract.

Example  4. MIA (monosodium iodoacetate -Induced Effects of Schizandrae Fructus Extract on Body Weight in Osteoarthritis-Induced Osteoarthritis

In order to confirm the effect of Schisandra chinensis leaf extract prepared in Example 1 on the weight load in MIA-induced osteoarthritis animal model, osteoarthritis-inducing substance MIA (diluted to a concentration of 60 mg / ml with 0.9% saline) Osteoarthritis was induced by injecting 50 μl into the right hind limb joint cavity of 7-week-old SD rats, followed by oral administration of Schisandra chinensis extract (150 and 300 mg / kg) once a day for a total of 21 days. Was measured. Indomethacin (1 mg / kg) was used as a positive control.

Hind limb weight load was measured using a foot weigher (Incapacitance tester, Linton instrument Co., UK, Ser No. 01/45/25). In osteoarthritis-induced rats in the tester's holder, the pain of the osteoarthritis stood on the normal foot without MIA, so the weight of both feet was unbalanced and the weight of the MIA-treated foot was relatively light. When measuring the weight of the foot, the weight (g) of both feet was measured without the SD rat's belly touching the sensor of the device.

Using the measured weight of the foot, the weight load rate (%) was calculated by the method of Equation 1 below. The weight load is the force supported by the foot, and in the normal case, the weight of both feet is balanced so that the weight load ratio of one foot is shown as 50%, but as the pain increases due to osteoarthritis, the weight load rate (%) of the hind limbs in which osteoarthritis is induced is increased. Lowers.

[Equation 1]

% Weight load = [weight of osteoarthritis-induced hind limb / (weight of hind limb of foot)] × 100

As a result, as shown in Figure 5 compared to the normal SD rat group was administered MIA compared to the weight-bearing rate (%) of the osteoarthritis-induced group was confirmed that the statistically significant decrease compared to the control group. In contrast, in the group administered with MIA and Schisandra chinensis extract, the weight loss rate (%) decreased statistically significantly from the 14th day.

Claims (10)

Schisandra chinensi s) health functional food composition for the prevention or improvement of arthritis containing leaf or stem extract as an active ingredient. The health functional food composition for preventing or improving arthritis according to claim 1, wherein the arthritis is osteoarthritis or gouty arthritis. The health functional food composition for preventing or improving arthritis according to claim 1, wherein the composition inhibits inflammatory cytokines and pain. The health functional food composition for preventing or improving arthritis according to claim 3, wherein the pain is pain caused by arthritis. The dietary supplement composition for preventing or improving arthritis according to claim 1, wherein the solvent of the Schizandra chinensis leaves or stem extract is water, a lower alcohol of C ₁ to C ₄ , or a mixture thereof. According to claim 1, wherein the composition is powder, granules, pills, tablets, capsules, candy, syrups and beverages, health functional food composition for the prevention or improvement of arthritis, characterized in that it is prepared in any one formulation. A pharmaceutical composition for the prevention or treatment of arthritis, containing Schizandra leaf or stem extract as an active ingredient. 8. The pharmaceutical composition for preventing or treating arthritis according to claim 7, wherein the arthritis is osteoarthritis or gouty arthritis. The pharmaceutical composition for preventing or treating arthritis of claim 7, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient. According to claim 7, wherein the composition is a pharmaceutical composition for the prevention or treatment of arthritis, characterized in that the formulation is made of any one of capsules, powders, granules, tablets, suspensions, emulsions, syrups and aerosols.

Images