Composition for prevention, amelioration or treatment of arthritis containing galla rhois extract as active ingredient

Abstract

The present invention relates to a composition for the prevention, amelioration or treatment of arthritis, containing a Galla rhois extract as an active ingredient. Specifically, in a collagen-induced arthritis animal model, upon administration of the Galla rhois extract, the expression levels of inflammatory cytokines (IL-1β, TNF-α, and IL-6) are reduced, the infiltration of inflammatory cells and the thickening of synovial membrane are suppressed in the ankle joint, cartilage loss of the articular surface is reduced, and bone destruction is improved. Therefore, the composition containing a Galla rhois extract as an active ingredient, of the present invention, can be effectively used as a material for the prevention, amelioration or treatment of arthritis, particularly rheumatoid arthritis.

Description

Composition for preventing, improving or treating arthritis containing extract of magnolia berry as an active ingredient

[0001] The present invention relates to a composition for preventing, improving or treating arthritis, containing an extract of Obaeja as an active ingredient.

[0002]

[0003] This work was carried out with the support of the Ministry of Science and ICT's major projects (Project Number: 1711177839).

[0004] Arthritis is a disease that causes inflammation and pain in the joints. It can be mainly divided into osteoarthritis and rheumatoid arthritis. Osteoarthritis, also called degenerative arthritis, occurs mainly in the elderly without a specific organic cause. If it progresses chronically, it can cause movement disorders such as difficulty walking due to deformation of the joint structure. Osteoarthritis is mainly caused by gradual damage to the articular cartilage or degenerative changes, which damage the bones and ligaments that form the joint, resulting in inflammation and pain. If we look at the mechanism, as osteoarthritis progresses, the production of inflammatory cytokines such as TNF-α (tumor necrosis factor-alpha) and IL-1β (Interleukin-1β) increases, and the secretion of matrix metalloproteinases (MMPs) such as collagenase and stromelysin increases, resulting in the destruction of articular cartilage. In addition, MMPs induce IL-1β, TNF-α, etc., which affect tissues such as muscles, tendons, and ligaments, causing severe pain. Here, MMPs are a major factor involved in cartilage damage, and can be divided into collagenases, gelatinases, stromelysins, membrane-type MMPs, and several other groups of MMPs.

[0005] Rheumatoid arthritis is an inflammatory disease characterized by polyarthritis, and autoimmune mechanisms are known to be the primary cause. Symptoms include inflammation of the synovial membrane, leading to the migration of macrophages, dendritic cells, T lymphocytes, and B lymphocytes into the synovial tissue. This leads to increased synovial fluid, swelling, and pain. If this inflammation persists, hyperplasia of the inflamed synovial tissue destroys bone and cartilage, deforming joint structures and causing movement disorders. Research has shown that in patients with rheumatoid arthritis, inflammatory cytokines induce the production of collagenases and neutral proteases in synovial fibroblasts and chondrocytes. These enzymes then destroy collagen and proteoglycans, ultimately destroying articular cartilage. When arthritis occurs, inflammation causes fever, swelling, and joint pain. If arthritis symptoms persist, cartilage and bone tissue are destroyed, leading to joint structural deformation and movement disorders. Therefore, there is a pressing need for medications that can suppress inflammation and pain, common symptoms of acute and chronic arthritis, while also improving or treating movement disorders, a chronic symptom. Furthermore, there is a pressing need for medications that are non-toxic to cartilage and synovial tissues, thereby protecting and maintaining joint structures.

[0006] On the other hand, Chinese gallnuts, also known as Rhus Galls in English or Galla Rhois or Galla Chinensis in Latin, are gallnuts formed by the parasitism of the aphid Schlechtendalia chinensis (Bell, 1851) of the Aphididae family on the leaves of plants of the Anacardiaceae family, such as Rhus chinensis Mill., Rhus potaninii Maxim., or Rhus punjabensis var. sinica (Diels) Rehder & E.H. Wilson. They are dried and used, and are also called Meretrix meretrix or Baichongcang.

[0007] As prior art related to arthritis, Korean Patent Registration No. 2475985 discloses a composition for preventing or treating rheumatoid arthritis containing a composite extract of rhubarb, gold, and coptis as an active ingredient, and Korean Patent Registration No. 1787082 discloses a pharmaceutical composition for treating rheumatoid arthritis containing an extract of balloon flower with an increased content of effective saponin as an active ingredient. However, there has been no disclosure yet regarding the 'composition for preventing, improving, or treating arthritis containing an extract of Chinese gallnuts as an active ingredient' of the present invention.

[0008] The present invention has been derived from the above requirements. The present invention provides a composition for preventing, improving, or treating arthritis containing an extract of Galla Rhois as an active ingredient. Specifically, in a collagen-induced arthritis animal model, when the extract of Chinese gallnuts is administered, the expression levels of inflammatory cytokines (IL-1β, TNF-α, and IL-6) are decreased, infiltration of inflammatory cells and thickening of the synovial membrane in the ankle joint are inhibited, cartilage loss on the joint surface is reduced, and bone destruction is improved. By confirming these effects, the present invention has been completed.

[0009] In order to solve the above problem, the present invention provides a health functional food composition for preventing or improving arthritis containing an extract of Obaeja as an effective ingredient.

[0010] In addition, the present invention provides a pharmaceutical composition for preventing or treating arthritis containing an extract of Obaeja as an active ingredient.

[0011] In addition, the present invention provides a feed additive for preventing or improving arthritis containing an extract of the quince plant as an effective ingredient.

[0012] In addition, the present invention provides a veterinary composition for preventing or treating arthritis containing an extract of the quince plant as an active ingredient.

[0013] In addition, the present invention provides a herbal medicine composition for preventing or treating arthritis containing an extract of Obaeja as an active ingredient.

[0014] The present invention relates to a composition for preventing, improving or treating arthritis containing a Galla Rhois extract as an active ingredient, and more specifically, in an animal model of arthritis induced by collagen, when a Galla Rhois extract is administered, the expression levels of inflammatory cytokines (IL-1β, TNF-α and IL-6) are reduced, infiltration of inflammatory cells and hypertrophy of the synovium in the ankle joint are suppressed, and cartilage loss on the joint surface is reduced and bone destruction is improved.

[0015] Figure 1 shows the results of confirming the effect of inhibiting articular cartilage loss and synovial hypertrophy by administering the Rhus chinensis extract of the present invention in an animal model of rheumatoid arthritis induced by collagen. CON is a normal group in which rheumatoid arthritis is not induced, CIA (collagen-induced arthritis) is a control group in which rheumatoid arthritis is induced by injecting collagen, and CIA+Rhododendron is a group in which Rhus chinensis extract (100 mg / kg) is administered to an animal model in which rheumatoid arthritis is induced by injecting collagen.

[0016] Figure 2 shows the results of confirming the change in the expression level of inflammatory cytokines (IL-1β, TNF-α, and IL-6) according to the administration of the Rhus chinensis extract of the present invention in an animal model of rheumatoid arthritis induced by collagen. CON is a normal group in which rheumatoid arthritis is not induced, CIA (collagen-induced arthritis) is a control group in which rheumatoid arthritis is induced by injecting collagen, and CIA+Rhod is a group in which Rhus chinensis extract (100 mg / kg) is administered to an animal model in which rheumatoid arthritis is induced by injecting collagen. # indicates that the expression level of inflammatory cytokines in the CIA control group (CIA) was statistically significantly increased compared to the normal group (CON), p<0.05. *, ** indicate that the expression of inflammatory cytokines in the CIA and Rhus officinalis extract administration group (CIA+Rho) was statistically significantly reduced compared to the CIA control group (CIA). * indicates p<0.05, and ** indicates p<0.01.

[0017] In order to achieve the purpose of the present invention, the present invention provides a health functional food composition for preventing or improving arthritis containing an extract of Galla Rhois as an effective ingredient.

[0018] The above arthritis is preferably rheumatoid arthritis, but is not limited thereto.

[0019] The above extract of the five-flowered quince may be prepared by a method including, but not limited to, the following steps:

[0020] 1) A step of extracting by adding an extraction solvent to the five-part extract;

[0021] 2) a step of filtering the extract of step 1); and

[0022] 3) A step of producing an extract by concentrating and drying the filtered extract of step 2) under reduced pressure.

[0023] In the above step 1), the extraction solvent is preferably water, a lower alcohol of C1 to C4, or a mixture thereof, more preferably water, but is not limited thereto.

[0024] In the above manufacturing method, the extraction of the gallnut can be performed using any conventional method known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction. The reduced pressure concentration in step 3) is preferably performed using a vacuum reduced pressure concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably performed using reduced pressure drying, vacuum drying, boiling drying, spray drying, or freeze drying, but is not limited thereto.

[0025] The above composition is preferably prepared in any one of the following dosage forms: powder, granules, pills, tablets, capsules, candies, syrups, and beverages, but is not limited thereto.

[0026] When the health functional food composition of the present invention is used as a food additive, the Gall extract can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention or improvement). Generally, when manufacturing a 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 below the above range, and since there is no problem in terms of safety, the active ingredient can be used in an amount exceeding the above range. There is no particular limitation on the type of food. Examples of foods to which the extract 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 all health functional foods in the conventional sense are included.

[0027] When the composition of the present invention is used as a health beverage, it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in conventional beverages. The natural carbohydrates mentioned above are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclotensin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame can 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 may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These components may be used independently or in mixtures. The proportion of these additives is not particularly important, but the composition of the present invention is typically selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.

[0028] In addition, the present invention provides a pharmaceutical composition for preventing or treating arthritis containing an extract of Galla Rhois as an active ingredient.

[0029] The above arthritis is preferably rheumatoid arthritis, but is not limited thereto.

[0030] The composition of the present invention is preferably prepared in any one formulation selected from among capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols, but is not limited thereto.

[0031] The composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the above-mentioned effective ingredient, and may be in various oral or parenteral dosage forms. When formulated, it is prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include capsules, powders, granules, tablets, pills, etc., and these solid preparations are prepared by mixing one or more compounds with at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, emulsions, syrups, and aerosols. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents can be propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases can include witepsol, macrogol, Tween 61, cacao butter, laurin butter, and glycerogelatin. For parenteral administration, it is preferable to select a method such as topical application to the skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine, epidural, or intracerebrovascular injection.

[0032] 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. The level of the effective amount may be determined based on factors including the type and severity of the patient's disease, the activity and sensitivity of the drug to the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, concurrently used drugs, 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 singly or in multiple doses. It is important to take all of the above factors into consideration and administer an amount that achieves the maximum effect with the minimum amount without causing side effects, and this can be easily determined by those skilled in the art.

[0033] The dosage of the composition of the present invention varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity, and the daily dosage is 0.01 to 2,000 mg / kg based on the amount of the Rhus chinensis extract, preferably 30 to 500 mg / kg, and more preferably 50 to 300 mg / kg, and can 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.

[0034] In addition, the present invention provides a feed additive for preventing or improving arthritis containing an extract of Galla Rhois as an effective ingredient.

[0035] The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act. The term "feed" in the present invention may mean any natural or artificial diet, meal, etc., or ingredients of the meal, which are intended for or suitable for animals to eat, ingest, and digest. The type of the feed is not particularly limited, and feed commonly used in the relevant technical field may be used. Non-limiting examples of the feed include plant feeds such as grains, roots, fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal, or grain by-products; and animal feeds such as proteins, inorganic substances, oils and fats, mineral substances, oils and fats, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0036] In addition, the present invention provides a veterinary composition for preventing or treating arthritis containing an extract of Galla Rhois as an active ingredient.

[0037] The veterinary composition of the present invention may further comprise suitable excipients and diluents according to conventional methods. Excipients and diluents that may be included in the veterinary composition 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.

[0038] The veterinary composition according to the present invention may further include fillers, anticoagulants, lubricants, wetting agents, flavoring agents, emulsifiers, preservatives, etc., and the veterinary composition according to the present invention may be formulated using methods well known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to an animal, and the formulation may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories, sterile injectable solutions, sterile topical preparations, etc.

[0039] The effective amount of the veterinary composition according to the present invention can be appropriately selected depending on the individual animal. This may be determined based on factors including the severity of the disease or condition, the individual's age, weight, health status, or sex, sensitivity to the active ingredient of the present invention, the route of administration, the duration of administration, other compositions combined with or used concurrently with the composition, and other factors well known in the fields of physiology and veterinary medicine.

[0040] In addition, the present invention provides a herbal medicine composition for preventing or treating arthritis containing an extract of Galla Rhois as an active ingredient.

[0041] The herbal medicine composition of the present invention means, but is not limited to, one manufactured according to a herbal medicine prescription.

[0042]

[0043] Hereinafter, the present invention will be described in detail by examples. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

[0044]

[0045] [Materials and Methods]

[0046] 1. Preparation of Galla Rhois extract

[0047] 16.05 kg of Rhizoma japonica (Jeongdo Herbal Medicine, Gyeonggi-do) was extracted three times for 5 hours each with 70 L of water, filtered through 3 layers of gauze and Whatman filter paper (Whatman No. 1), concentrated under reduced pressure in a rotary vacuum evaporator, and then freeze-dried. The total recovery amount of Rhizoma japonica extract was 7173.55 g. The Rhizoma japonica extract was stored in the refrigerator, completely dissolved in sterilized saline solution before the experiment, and filtered through a membrane filter (0.45 μm) to use as the experimental drug. The remaining sample was stored frozen (-20°C).

[0048]

[0049] 2. Animals

[0050] Experimental animals were 7- to 8-week-old male DBA / 1J mice (20-25 g) purchased from Orient Bio Co., Ltd. (Gyeonggi-do, Korea). All experimental animals were provided with unrestricted access to solid food and water, and were housed under a constant temperature (23±3°C), humidity (50±10%), and 12-h light-dark cycle.

[0051]

[0052] 3. Production of collagen-induced arthritis animals

[0053] For collagen-induced arthritis (CIA), 2 mg / mL of bovine type II collagen was dissolved in 0.01 N acetic acid at 4°C overnight, mixed well with an equal volume (w / v) of Freund's complete adjuvant (Chondrex, SA, USA), and 0.1 mL was intradermally injected into the buttocks (the area where the tail connects to the body) of mice. Fourteen days later, arthritis was induced by subcutaneously injecting 0.1 mL of the collagen solution prepared in the same manner into the tarsal joint of the hind legs of mice mixed with an equal volume of Freund's incomplete adjuvant (FIA).

[0054] The experimental group consisted of 10 mice per group, divided into a normal control group (CON) that did not induce arthritis, a control group that induced arthritis (CIA), and an experimental group that induced arthritis and was administered the extract of Rhus verniciflua.

[0055] Drug administration was performed by orally administering 100 mg / kg of ethanol extract of Gaebaekja once a day at a set time for 28 days from day 28 to day 48 after CIA induction. On day 49, all experimental animals were sacrificed to evaluate the degree of improvement in arthritis.

[0056]

[0057] 4. Tissue specimen preparation and staining

[0058] On the 49th day of the experiment, all experimental animals were sacrificed, and the area between the distal tibia and the distal metatarsal was removed to include the right ankle joint, immersed in 10% (v / v) neutral formalin solution (formaldehyde), fixed at 4℃ for 24 hours, and then decalcified by immersion in 20% (v / v) formic acid for 3 days. The decalcified tissue was washed for 24 hours to remove any remaining formalin, dehydrated in increasing concentrations of 50% (v / v) alcohol to absolute alcohol, and replaced with xylene. After that, paraffin was infiltrated and embedded to create paraffin blocks, and sliced ​​into 5 ㎛ thick sections using a rotary microtome and placed on slides to create tissue specimens.

[0059] The paraffin tissue specimens were deparaffinized in the order of xylene, absolute alcohol, and 50% alcohol, and then stained with H&E (Haematoxylin & eosin) staining reagent to observe the morphological changes of the joint tissue under an optical microscope.

[0060]

[0061] 5. ELISA (Enzyme-linked immunosorbent assays)

[0062] IL-1β, IL-6, and TNF-α contents were determined from mouse paw homogenate supernatants using Duoset ELISA kits from R&D Systems (Minneapolis, MN, USA) according to the manufacturer's protocol.

[0063]

[0064] 6. Statistical testing

[0065] All experimental results were repeated three times and expressed as the mean and standard deviation (mean±SD). Statistical significance was verified using one-way ANOVA and Tukey's test in the GraphPad Prism 5.0 analysis program. If the p value was less than 0.05, it was considered statistically significant.

[0066]

[0067] Example 1. Morphological changes in joint tissues following administration of the extract of the present invention in an animal model of arthritis induced by collagen.

[0068] To investigate the effect of extracts of Galla japonica on damage to the ankle joints in mice with rheumatoid arthritis induced by collagen injection, joint tissues were stained with H&E.

[0069] As a result, as shown in Fig. 1, in the control group (CIA) in which rheumatoid arthritis was induced by injecting collagen compared to the normal group (CON), significant infiltration of polymorphic leukocytes and lymphocytes along with edema around the joints was observed, and significant cartilage loss (bone erosion) of the joint surface was observed along with synovial inflammation and hypertrophy.

[0070] In contrast, in the group administered with the extract of the present invention (CIA + Galla), infiltration of inflammatory cells and hypertrophy of the synovium were suppressed, cartilage loss on the joint surface was reduced, and bone destruction was improved compared to the control group (CIA) in which rheumatoid arthritis was induced.

[0071]

[0072] Example 2. Changes in the content of inflammatory cytokines according to treatment with the extract of the gallnut of the present invention in an animal model of arthritis induced by collagen.

[0073] To investigate the effect of the extract of Rhus verniciflua on the contents of IL-1β, TNF-α, and IL-6, which are inflammatory cytokines produced in large quantities in mice with rheumatoid arthritis induced by collagen injection, the contents of IL-1β, TNF-α, and IL-6 in the joints were measured using the ELISA method.

[0074] As a result, as shown in Fig. 2, the content of inflammatory cytokines was significantly increased in the control group (CIA) in which rheumatoid arthritis was induced by injection of collagen compared to the normal group (CON), and in the group administered with the extract of the gall tree of the present invention, IL-1β, TNF-α, and IL-6 were significantly decreased compared to the control group in which rheumatoid arthritis was induced.

Claims

1. A health functional food composition for preventing or improving arthritis containing Galla Rhois extract as an active ingredient.

2. A health functional food composition for preventing or improving arthritis, characterized in that the arthritis in paragraph 1 is rheumatoid arthritis.

3. A health functional food composition for preventing or improving arthritis, characterized in that the extraction solvent of the extract in paragraph 1 is water, a C1-C4 lower alcohol, or a mixture thereof.

4. A health functional food composition for preventing or improving arthritis, characterized in that the composition is manufactured in any one formulation selected from powder, granules, pills, tablets, capsules, candy, syrup, and beverage in the first paragraph.

5. A pharmaceutical composition for the prevention or treatment of arthritis containing an extract of Galla Rhois as an active ingredient.

6. A pharmaceutical composition for preventing or treating arthritis, characterized in that in paragraph 5, the composition further comprises a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient.

7. A pharmaceutical composition for preventing or treating arthritis, characterized in that the composition is prepared in any one formulation selected from among capsules, powders, granules, tablets, suspensions, emulsions, syrups, and aerosols in accordance with paragraph 5.

8. Feed additive for preventing or improving arthritis containing Galla Rhois extract as an active ingredient.

9. A veterinary composition for the prevention or treatment of arthritis containing an extract of Galla Rhois as an active ingredient.

10. A herbal medicine composition for the prevention or treatment of arthritis containing an extract of Galla Rhois as an active ingredient.

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