KR20210137969A - Composition for preventing and treating osteoarthritis comprising curcumin and polyunsaturated fattyacids - Google Patents

Abstract

Curcumin and polyunsaturated fatty acids omega 3 fatty acids or omega 6 fatty acids are treated in combination to relieve pain caused by arthritis, have a cartilage protection effect, and have an effect of inhibiting inflammatory cytokines and cartilage destruction. Therefore, curcumin and polyunsaturated fatty acids omega 3 fatty acids or omega 6 fatty acids can be usefully used as a composition for preventing or treating arthritis or osteoporosis.

Description

Composition for the prevention and treatment of arthritis containing curcumin and polyunsaturated fatty acids as active ingredients

The present invention relates to the prevention or treatment effect of arthritis by the combined treatment of curcumin and polyunsaturated fatty acids.

The human body consists of about 200 joints. Joints are the places where bones meet. Joints are composed of cartilage, joint capsule, synovial membrane, ligaments, tendons, muscles, etc. so that bones can move smoothly between bones, and they serve to absorb shocks caused by movement. Inflammatory diseases appearing in these joints are chronic joint rheumatism, which is understood to be caused by autoimmunity, infectious arthritis caused by bacterial infection, degenerative arthritis in which degeneration or destruction of articular cartilage or bone due to various causes, and degenerative changes in connective tissue. It can be broadly classified into crystalline arthritis, in which soluble metabolites are deposited as crystals in the connective tissue around the joint. In degenerative arthritis, that is, osteoarthritis, degeneration such as aging occurs in the chondrocytes constituting the joint, and the synthesis of type II collagen and proteoglycan, which are substrate materials of the joint, is inhibited in the chondrocytes, and at the same time, As inflammatory cytokines such as interleukin-1 and tumor necrosis factor-α are produced, the synthesis of matrix metalloproteinase (MMP) and It is a disease caused by destruction of joint tissue due to increased activity in joint cells. In addition, arthritis is further exacerbated by the induced synthesis of more MMPs due to the production of nitric oxide by inflammatory cytokines and the production of self-amplifying cytokines by the produced nitric oxide, thereby promoting the decomposition of the joint matrix. At the same time, inflammatory cytokines increase the production of prostaglandin E2, a lipid metabolite, to induce an inflammatory response in arthritis. Osteoarthritis, also called degenerative arthritis, is a type of arthritis that occurs mainly in old age without a specific organic cause. Osteoarthritis is mainly caused by damage to the bones and ligaments that make up the joint due to gradual damage or degenerative changes in the articular cartilage, resulting in inflammation and pain. When osteoarthritis progresses, the production of inflammatory cytokines such as TNF-α and IL-1 increases, and the secretion of MMPs such as collagenase and stromelysin increases, leading to the destruction of articular cartilage. will be done In addition, MMPs induce IL-1 and TNF-α, which in turn affects tissues such as muscles, tendons, and ligaments, causing severe pain.

MMPs are proteolytic enzymes that destroy the matrix components of bone and cartilage, and are expressed in cartilage tissues stimulated by inflammatory cytokines during an inflammatory disease state and their activity increases. MMPs consist of at least 21 enzymes, including collagenase (MMP-1,8,13), stromelysin (MMP-3,10,11), gelatinase (MMP-2,9) and matrix type-1 metalloproteinase (MMP-14). ) is subdivided into subgroups.

Rheumatoid arthritis and degenerative arthritis are characterized by infiltration of inflammatory cells into the joint synovial tissue, which is mediated by chemokines. Chemokines such as monocyte chemoattractant protein-1 (MCP-1) are expressed in the synovial tissue of arthritis, and these are known to be produced in synovial fibroblasts. Excess MCP-1 produced by arthritis invites monocytes and macrophages to the site of inflammation, and by activating these cells, it promotes the production of inflammatory cytokines, thereby exacerbating inflammation.

Currently, for the treatment of osteoarthritis, drugs such as acetaminophen, tramadol, nonsteroidal anti-inflammatory drugs (NSAIDs), diaceline, glucosamine, etc. are used, but only to temporarily relieve the pain of osteoarthritis. Accordingly, there is a need for a treatment method capable of effectively alleviating pain while treating osteoarthritis.

On the other hand, curcumin (curcumin) is _{a compound with a chemical formula of C 21} H ₂₀ O ₆ and a molecular weight of 368.38, extracted from the root of Curcuma longa Linn (Zingiberaceae), a plant belonging to the Ginger family of East India, and widely used in Indian food. It is a yellow spice containing all polyphenols and is a curcuminoid of turmeric. In food additives, it is used as a yellow colorant and as a spice. Curcumin has anti-tumor, antioxidant, anti-amyloid and anti-inflammatory actions. It has been reported that it inhibits gastric cancer formation induced by methyl-N'-nitro-N-nitrosoguanidin.

Korean Patent No. 0626358

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating arthritis.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating osteoporosis.

Another object of the present invention is to provide a food composition for preventing or improving arthritis.

In addition, it is an object of the present invention to provide a food composition for preventing or improving osteoporosis.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating arthritis comprising curcumin and polyunsaturated fatty acids as active ingredients.

In addition, the present invention provides a pharmaceutical composition for preventing or treating osteoporosis comprising curcumin and polyunsaturated fatty acids as active ingredients.

In addition, the present invention provides a food composition for preventing or improving arthritis comprising curcumin and polyunsaturated fatty acids.

In addition, the present invention provides a food composition for preventing or improving osteoporosis comprising curcumin and polyunsaturated fatty acids.

According to the present invention, curcumin and polyunsaturated fatty acids omega 3 fatty acids or omega 6 fatty acids are treated in combination to relieve pain due to arthritis, have a cartilage protection effect, and inhibit inflammatory cytokines and cartilage destruction. It can be usefully used as a composition for preventing or treating arthritis or osteoporosis.

1 is a diagram confirming the pain suppression effect of curcumin and omega 3 in an animal model of osteoarthritis as paw withdrawal latency (PWL).
2 is a diagram confirming the pain suppression effect of curcumin and omega 3 in an animal model of osteoarthritis as a paw withdrawal threshold (PWT).
3 is a diagram confirming the pain suppression effect of curcumin and omega 3 in an animal model of osteoarthritis by weight bearing.
4 is a view confirming the chondroprotective effect of curcumin and omega 3 in an animal model of osteoarthritis through bone volume.
Figure 5 is a diagram confirming the chondroprotective effect of curcumin and omega 3 in an animal model of osteoarthritis through safranin O (Safranin O) staining, OARSI score, Mankin score, number of cartilage cells, cartilage destruction and tidemark integrity. am.
6 is a diagram confirming the change in MCP-1 expression caused by curcumin and omega-3.
7 is a diagram confirming the pain suppression effect of curcumin and linoleic acid in an animal model of osteoarthritis as paw withdrawal latency (PWL).
8 is a diagram confirming the pain suppression effect of curcumin and linoleic acid in an animal model of osteoarthritis as a paw withdrawal threshold (PWT).
9 is a diagram illustrating the weight-bearing effect of curcumin and linoleic acid to suppress pain in an animal model of osteoarthritis.
10 is a view confirming the chondroprotective effect of curcumin and linoleic acid in an animal model of osteoarthritis through bone volume.
11 is a diagram confirming the chondroprotective effect of curcumin and linoleic acid in an animal model of osteoarthritis through safranin O staining, OARSI score, Mankin score, number of cartilage cells, cartilage destruction and tidemark integrity. .
12 is a diagram confirming the expression change of MMP1, MMP3 and MMP13 by curcumin, omega-3 and linoleic acid:
Nil: control not treated with IL-1β.

Hereinafter, the present invention will be described in detail as an embodiment of the present invention with reference to the accompanying drawings. However, the following embodiments are presented as examples of the present invention, and when it is determined that detailed descriptions of well-known techniques or configurations known to those skilled in the art may unnecessarily obscure the gist of the present invention, the detailed description may be omitted, and , the present invention is not limited thereby. Various modifications and applications of the present invention are possible within the scope of equivalents interpreted therefrom and the description of the claims to be described later.

In addition, the terms used in this specification are terms used to properly express the preferred embodiment of the present invention, which may vary according to the intention of a user or operator, or customs in the field to which the present invention belongs. Accordingly, definitions of these terms should be made based on the content throughout this specification. Throughout the specification, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

All technical terms used in the present invention, unless otherwise defined, have the meaning as commonly understood by one of ordinary skill in the art of the present invention. In addition, although preferred methods and samples are described herein, similar or equivalent ones are also included in the scope of the present invention. The contents of all publications herein incorporated by reference are incorporated herein by reference.

In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating arthritis comprising curcumin and polyunsaturated fatty acids as active ingredients.

In one embodiment, the polyunsaturated fatty acid may be an omega 3 (Omega3) fatty acid, an omega 6 fatty acid or an omega 3 fatty acid and an omega 6 fatty acid.

In one embodiment, the omega-3 fatty acid is made in the form of free fatty acid (FFA) in the process of increasing the content of omega-3 by refining refined fish oil containing omega-3 and other fatty acids to selectively purify only omega-3 It may be in the form of omega 3 EE with an ethyl ester group attached to the end of the FFA to increase stability.

In one embodiment, the omega 3 fatty acid is eicosapentaenoic acid (EPA) or a linear or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, docosahexaenoic acid (docosahexaenoic acid) , DHA) or straight-chain or branched C1-C6 alkyl esters thereof, triglycerides or phospholipids thereof, alpha-linolenic acid (ALA) or straight-chain or branched C1-C6 alkyl esters thereof; of triglycerides or phospholipids, stearidonic acid (SDA) or straight-chain or branched C1-C6 alkyl esters thereof, triglycerides or phospholipids thereof, and eicosatetraenoic acid (ETA) Or it may be any one or more selected from the group consisting of linear or branched C1-C6 alkyl esters thereof, triglycerides or phospholipids thereof.

In one embodiment, the omega-6 fatty acid is linoleic acid or a linear or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, arachidonic acid or a straight-chain or branched C1- thereof C6 alkyl esters, triglycerides or phospholipids thereof, 7,10-hexadecadienoate, 10-hexadecadienoate, or straight-chain or branched C1-C6 selected from the group consisting of alkyl esters, triglycerides or phospholipids thereof, and dihomo-gamma-linolenic acid (DHGLA) or straight or branched C1-C6 alkyl esters thereof, triglycerides or phospholipids thereof It may be any one or more.

In one embodiment, curcumin and omega 3 and/or linoleic acid may be included as active ingredients.

In one embodiment, curcumin and omega 3 may be mixed in a weight ratio of 10:11, curcumin and linoleic acid may be mixed in a weight ratio of 5:11, and curcumin, omega 3 and linoleic acid may be mixed in a weight ratio of 10:11:22 can be mixed with

In one embodiment, the arthritis is senile arthritis, degenerative arthritis, autoimmune arthritis, osteoarthritis, rheumatoid arthritis, spondyloarthropathy, ankylosing spondylitis, psoriatic arthritis, gout, bacterial arthritis, juvenile rheumatoid arthritis, lupus, scleroderma, multiple sclerosis, fibromyalgia , polymyositis, dermatomyositis, Behcet's disease, Reiter's syndrome, Lyme arthritis, adhesive capsulitis, frozen shoulder, tendon synovitis, elbow capsulitis, de Quervain's tendon synovitis, recurrent rheumatism, polymyalgia rheumatica and adult-type Still's disease It may be any one or more selected from the group.

In one embodiment, the composition of the present invention can reduce cartilage destruction by inhibiting the expression of MCP-1, a chemokine that aggravates inflammation caused by arthritis, and suppressing the expression of cartilage-destructive factors MMP1, MMP3 and MMP13.

In one embodiment, the composition of the present invention comprises curcumin in a concentration of 2 mg/kg to 5000 mg/kg, omega-3 fatty acids in a concentration of 2.2 mg/kg to 5500 mg/kg, and omega-6 fatty acids in a concentration of 4.4 mg/kg It may be included in a concentration of kg to 11000 mg/kg.

In one embodiment, the composition of the present invention may be a composition for alleviating pain caused by arthritis.

In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating osteoporosis comprising curcumin and polyunsaturated fatty acids as active ingredients.

In one embodiment, the polyunsaturated fatty acid may be omega 3 or linoleic acid.

The curcumin according to the present invention may be used in the form of a salt, preferably a pharmaceutically acceptable salt. The salt is preferably an acid addition salt formed with a pharmaceutically acceptable free acid, and an organic acid and an inorganic acid may be used as the free acid. The organic acid is not limited thereto, but citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, metasulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid and aspartic acid. In addition, the inorganic acid includes, but is not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid. The curcumin compound according to the present invention may be isolated from nature or prepared by a chemical synthesis method known in the art.

The pharmaceutical composition of the present invention may further include an adjuvant. The adjuvant may be used without limitation as long as it is known in the art, and for example, Freund's complete adjuvant or incomplete adjuvant may be further included to increase the immunity thereof.

As used herein, the term "treatment" means, unless otherwise stated, the disease or condition to which the term applies, or one or more symptoms of the disease or disorder, which reverses, ameliorates, inhibits the progression, or means to prevent, and the term treatment as used herein refers to the act of treating. Accordingly, treatment or therapy for immune diseases such as osteoarthritis in mammals may include one or more of the following:

(1) inhibit the growth of, ie, arrest the development of, an immune disease;

(2) prevent the spread of immune diseases, ie, prevent metastasis;

(3) alleviating immune disorders;

(4) prevent recurrence of immune diseases; and

(5) alleviating symptoms of immune diseases (palliating)

As used herein, the term "mammal" refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.

A composition is indicated to be "pharmaceutically or physiologically acceptable" if the recipient animal can tolerate administration of the composition, or if administration of the composition to that animal is suitable. When the amount administered is physiologically important, the agent can be said to have been administered in a "therapeutically effective amount". An agent is physiologically meaningful if the presence of the agent results in a physiologically detectable change in the recipient patient.

The therapeutically effective amount of the composition of the present invention may vary depending on several factors, for example, the administration method, the target site, the condition of the patient, and the like. Therefore, when used in the human body, the dosage should be determined as an appropriate amount in consideration of both safety and efficiency. It is also possible to estimate the amount used in humans from the effective amount determined through animal experiments. These considerations in determining effective amounts are found, for example, in Hardman and Limbird, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; and E.W. Martin ed., Remington's Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and not to cause side effects, and the effective dose level is determined by the patient's Health status, disease type, severity, drug activity, drug sensitivity, administration method, administration time, administration route and excretion rate, treatment period, factors including drugs used in combination or concurrently, and other factors well known in the medical field can be determined according to 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 multiple times. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.

The compositions of the present invention may also include carriers, diluents, excipients or combinations of two or more commonly used in biological agents. A pharmaceutically acceptable carrier is not particularly limited as long as it is suitable for in vivo delivery of the composition, for example, Merck Index, 13th ed., Merck & Co. Inc. Compounds described in , saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and one or more of these components can be mixed and used, and if necessary, other antioxidants, buffers, bacteriostats, etc. Conventional additives may be added. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to formulate into injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules or tablets. Furthermore, it can be preferably formulated according to each disease or component using an appropriate method in the art or a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990).

The pharmaceutical composition of the present invention may be formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, or sterile injection solutions according to conventional methods, respectively. have.

As used herein, the term “pharmaceutically acceptable” refers to exhibiting non-toxic properties to cells or humans exposed to the composition.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive includes starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate. , lactose, mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, lead carnauba, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, Calcium stearate, sucrose, dextrose, sorbitol and talc and the like can be used. The pharmaceutically acceptable additive according to the present invention is preferably included in an amount of 0.1 to 90 parts by weight based on the composition, but is not limited thereto.

As used herein, the term "administration" means providing a predetermined substance to a patient by any suitable method, and parenteral administration (eg, intravenous, subcutaneous, intraperitoneal or topical administration according to a desired method) ) or oral administration, and the dosage varies according to the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease.

The composition of the present invention may be administered parenterally (eg, intravenously, subcutaneously, intraperitoneally or topically) or orally according to a desired method, and the dosage may vary depending on the subject's age, weight, sex, physical condition, etc. is selected taking into account. It is self-evident that the concentration of the active ingredient included in the pharmaceutical composition can be variously selected depending on the subject, and is preferably included in the pharmaceutical composition at a concentration of 0.01 to 5,000 μg/ml. If the concentration is less than 0.01 μg/ml, pharmaceutical activity may not appear, and if it exceeds 5,000 μg/ml, it may be toxic to the human body.

The pharmaceutical composition of the present invention may be formulated in various oral or parenteral dosage forms. Formulations for oral administration include, for example, tablets, pills, hard, soft capsules, solutions, suspensions, emulsifiers, syrups, granules, and the like. crose, mannitol, sorbitol, cellulose and/or glycine), lubricants (eg silica, talc, stearic acid and its magnesium or calcium salts and/or polyethylene glycol). In addition, the tablet may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidine, and optionally starch, agar, alginic acid. or disintegrants such as sodium salts thereof or effervescent mixtures and/or absorbents, coloring, flavoring and sweetening agents. The formulation may be prepared by conventional mixing, granulating or coating methods. In addition, a representative formulation for parenteral administration is an injection formulation, and water, Ringer's solution, isotonic saline, or suspension may be mentioned as a solvent for the injection formulation. The sterile, fixed oil of the injectable preparation may be used as a solvent or suspending medium, and any non-irritating fixed oil including mono- and di-glycerides may be used for this purpose. In addition, the injection preparation may use a fatty acid such as oleic acid.

In the present invention, the term "prevention" refers to any action that inhibits or delays the occurrence, spread and recurrence of arthritis or osteoporosis by administration of the pharmaceutical composition according to the present invention.

In one aspect, the present invention relates to a food composition for preventing or improving arthritis comprising curcumin and polyunsaturated fatty acids as active ingredients.

In one embodiment, the polyunsaturated fatty acid may be an omega 3 fatty acid or an omega 6 fatty acid, more preferably omega 3 and/or linoleic acid.

In one aspect, the present invention relates to a food composition for preventing or improving osteoporosis comprising curcumin and polyunsaturated fatty acids as active ingredients.

In one embodiment, the polyunsaturated fatty acid may be an omega 3 fatty acid or an omega 6 fatty acid, more preferably omega 3 and/or linoleic acid containing a large amount of DHA.

When the composition of the present invention is used as a food composition, the composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. In addition to the active ingredient, the composition may contain a food additive that is pharmaceutically acceptable, and the mixing amount of the active ingredient may be suitably determined according to the purpose of use (prevention, health or therapeutic treatment).

The term "food supplement additive" used in the present invention refers to a component that can be added to food as an auxiliary, and is added to the manufacture of health functional food of each formulation, and those skilled in the art can appropriately select and use it. Examples of food supplement additives include various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverages, etc., but the above examples are not limited to the type of food supplement additive of the present invention.

The food composition of the present invention may include a health functional food. The term "health functional food" as used in the present invention refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. using raw materials or ingredients useful in the human body. Here, the term 'functionality' refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the ordinary technical field, and at the time of the preparation, it can be prepared by adding raw materials and components commonly added in the conventional technical field. In addition, if the formulation of the health functional food is also recognized as a health functional food, it can be prepared without limitation. The composition for food of the present invention can be prepared in various forms, and unlike general drugs, it has the advantage that there are no side effects that may occur during long-term administration of the drug using food as a raw material, and has excellent portability, and the present invention health functional food can be taken as an adjuvant to enhance the effect of arthritis or osteoporosis treatment.

In addition, there is no limitation on the type of health food in which the composition of the present invention can be used. In addition, the composition comprising the extract of the present invention or a fraction thereof as an active ingredient can be prepared by mixing known additives with other suitable auxiliary ingredients that may be contained in health functional foods according to the selection of those skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages and There are vitamin complexes and the like, and it can be prepared by adding the extract according to the present invention as a main component to juice, tea, jelly, juice, and the like.

Since the composition of the present invention is made from natural materials, side effects may be less than that of general synthetic compounds even when used as a pharmaceutical composition or a food composition, so it can be safely included in pharmaceutical compositions and health functional foods to be usefully used.

In one aspect, the invention relates to a kit comprising a composition of the invention in one or more unit dosage forms.

In one embodiment, the dosage forms described herein may be packaged in a bottle or as a blister pack with instructions for use of the dosage form. For example, instructions may be provided as a package insert or may be provided directly on a blister pack, label affixed to the bottle, or on a secondary packaging material from which the blister pack or bottle was provided to a human subject. Instructions may include, for example, the number of doses, administration of the dosage form administered with or without food, the active ingredients constituting the dosage form, and cardiovascular conditions or disorders that would benefit from administration of the dosage form.

In one embodiment, curcumin and an omega 3 fatty acid and/or an omega 6 fatty acid may be administered simultaneously or sequentially.

The present invention will be described in more detail through the following examples. However, the following examples are only for specifying the contents of the present invention, and the present invention is not limited thereto.

Example 1. Construction of an animal model of osteoarthritis

In order to produce an osteoarthritis animal model, 5-week-old male Wistar rats weighing 200-250 g were bred at a temperature of 21-22 ° C in a light-dark cycle at 12 hour intervals, and sterilized water and feed were supplied. brought up Then, for osteoarthritis induction, monosodium iodoacetate (MIA, Sigma, ST. Louis, MO) was administered at a dose of 3 mg/50 μl to the right knee of a rat to induce osteoarthritis. MIA was administered by dissolving it in physiological saline.

Example 2. Confirmation of osteoarthritis treatment effect of curcumin and omega 3

2-1. Check the pain relief effect

Vehicle (untreated control group), curcumin 100mg/kg, curcumin 100mg/kg+ omega 3 EE (free fatty acid (FFA) form in the process of increasing the content of omega-3 by refining refined fish oil containing omega-3 and other fatty acids After selectively refining only omega-3 with FFA, 110 mg/kg (formed with an ethyl ester group attached to the end of FFA) and 30 mg/kg of arthritis treatment Celecoxib (positive control) were orally administered 6 times a week, respectively, 2 After one day, pain was measured using a Dynamic plantat aesthsiometer (Ugo Basile, Comerio, Italy). To measure the pain of the machine, a net plate was placed on the measuring machine, a rat was placed in an acrylic animal frame on it, and then the drug was stabbed into the right foot with the measuring machine. The pain measurement graph was drawn by measuring the time it takes for the machine to automatically withdraw the paw after the stab (s, seconds) and the amount of weight applied to the paw withdrawal threshold (g). . In addition, an incapacitance tester was used to compare and measure the force applied to the left and right feet of the rat. The rat was placed in a plastic chamber, and the left and right feet of the rats were placed on each plate on a separate plate of the incapacitance tester, and the average value of the force applied to each paw was obtained for 3 seconds. The weight bearing (%) obtained in this way was substituted into Equation 1 below to obtain a result.

As a result, it was found that the pain of the experimental groups was alleviated compared to the group administered only MIA (control group, vehicle), and the group administered with omega 3 and curcumin together showed a significant effect on pain relief than the conventional osteoarthritis treatment, It was found that the pain relief effect persisted even after the elapsed time after administration ( FIGS. 1 and 2 ). In addition, the weight-bearing effect of the group administered with omega 3 and curcumin was the most excellent (Fig. 3).

2-2. Check the cartilage protection effect

After administering the drugs as in Example 2-1, in order to confirm the degree of cartilage damage in each administration group, after collecting the femur and tibia of the rat, the bone volume (%) was measured by micro-CT. The degree of cartilage destruction was confirmed by analysis. In addition, the extracted and isolated joints were subjected to Safranin O (Safranin O) staining, a staining method that can determine the degree of cartilage destruction, to observe the shape of cartilage, OARSI score, Mankin score, number of cartilage cells, Cartilage destruction and tidemark integrity were analyzed.

As a result, it was observed that the reduction of inflammatory cells in the cartilage and the destruction of cartilage were suppressed, and the maintenance degree of cartilage was well maintained close to that of normal tissue in the group administered with the combination of omega 3 and curcumin. Specifically, the group administered with omega 3 and curcumin showed significantly higher bone volume even in the conventional osteoarthritis treatment group (Fig. 4), and when omega 3 and curcumin were administered together, OARSI score, Mankin score, and cartilage Cell number, cartilage destruction and tidemark integrity were all significantly reduced ( FIG. 5 ). Through this, it was found that when omega 3 and curcumin were administered together, cartilage destruction was significantly inhibited.

2-3. Confirmation of inflammatory cytokine control effect

Real-time PCR was performed to determine the effect of omega-3 + curcumin combination treatment on the expression of monocyte chemoattractant protein-1 (MCP-1), a chemokine that exacerbates inflammation by promoting the production of inflammatory cytokines by overexpression in synovial tissue due to arthritis. investigated. Specifically, in an in vitro experiment, human-derived chondrocytes were pre-treated for 24 hours to stabilize the cells, and then stimulated with IL-1β to make them in a state similar to osteoarthritis cartilage, and vehicle, omega 3 (omega 3 is in OIL state, so cells are treated Since it is difficult to treat the cells using DHA, the main component of omega 3), 50 uM + 100 uM of curcumin and 10 uM of Celecoxib were treated, respectively. After culturing the cells for one day, RNA was extracted, cDNA was synthesized, and Realtime PCR analysis was performed.

As a result, it was shown that omega 3 + curcumin significantly inhibited the expression of MCP-1 than Celecoxib, a conventional treatment for arthritis ( FIG. 6 ).

Example 3. Confirmation of the osteoarthritis treatment effect of curcumin and linoleic acid

3-1. Check the pain relief effect

As in Example 2-1, vehicle (untreated control group), curcumin 100mg/kg, curcumin 100mg/kg+linoleic acid 220mg/kg, and arthritis treatment Celecoxib 30mg/kg (positive control) were orally administered 2 days after each oral administration Pain was measured using a plantat aesthsiometer (Ugo Basile, Comerio, Italy). To measure the pain of the machine, a net plate was placed on the measuring machine, a rat was placed in an acrylic animal frame on it, and then the drug was stabbed into the right foot with the measuring machine. I drew a graph of pain measurement by measuring the time it takes for the machine to automatically release the paw after stabbing (paw withdrawal latenvy) (s, seconds) and how much weight it takes to release the paw (paw withdrawal threshold) (g) . In addition, an incapacitance tester was used to compare and measure the force applied to the left and right feet of the rat. The rat was placed in a plastic chamber, and the left and right feet of the rats were placed on each plate on a separate plate of the incapacitance tester, and the average value of the force applied to each paw was obtained for 3 seconds. The weight bearing (%) obtained in this way was substituted in Equation 1 to obtain a result.

As a result, it was found that the pain of the experimental groups was alleviated compared to the group administered with MIA alone (control group, vehicle), and the group administered with linoleic acid and curcumin together showed a pain relief effect similar to that of the conventional osteoarthritis treatment, It was found that the pain relief effect persisted even after the lapse of time ( FIGS. 7 and 8 ). In addition, the weight-bearing effect of the group administered with linoleic acid and curcumin was similar to that of the conventional treatment for osteoarthritis (FIG. 9).

3-2. Check the cartilage protection effect

After administering the drugs as in Example 3-1, in order to confirm the degree of cartilage damage in each administration group, after collecting the femur and tibia of the rat, the bone volume (%) was measured by micro-CT. The degree of cartilage destruction was confirmed by analysis. In addition, the extracted and isolated joints were subjected to Safranin O (Safranin O) staining, a staining method that can determine the degree of cartilage destruction, to observe the shape of cartilage, OARSI score, Mankin score, number of cartilage cells, Cartilage destruction and tidemark integrity were confirmed as in Example 2-2.

As a result, the group administered with linoleic acid and curcumin showed significantly higher bone volume than the group administered with the conventional osteoarthritis treatment group (FIG. 10), and when linoleic acid and curcumin were administered together, the OARSI score, Mankin score, number of chondrocytes, and cartilage destruction and tidemark integrity were both significantly reduced ( FIG. 11 ). Through this, it was found that when linoleic acid and curcumin were administered together, cartilage destruction was significantly inhibited.

Example 4. Confirmation of osteoarthritis treatment effect of curcumin, omega 3 and linoleic acid

* It was confirmed by real-time PCR whether the combined treatment of curcumin, omega-3 and linoleic acid had an effect on the expression of MMP1 (metalloproteinase1), MMP3 and MMP13, which are catabolic molecules related to cartilage destruction. Specifically, in an in vitro experiment, human-derived chondrocytes were pre-treated for 24 hours to stabilize the cells, and then stimulated with IL-1β to make them in a state similar to osteoarthritic cartilage. and then incubated for 24 hours. Then, RNA was obtained from the cells and the expression of MMP1, MMP3 and MMP13 was observed by real-time PCR.

As a result, in the case of the group treated with omega 3 + curcumin + linoleic acid, the expression of MMP1, MMP3 and MMP13 was significantly reduced. decreased (Fig. 12). Through this, it was found that the combined treatment of omega 3, curcumin, and linoleic acid of the present invention reduced catabolism and exhibited an excellent effect on cartilage protection and regeneration.

Claims (9)

As a pharmaceutical composition for preventing or treating arthritis, comprising curcumin, omega 3 fatty acids and linoleic acid as active ingredients,
The omega 3 fatty acid is eicosapentaenoic acid (EPA) or a linear or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, docosahexaenoic acid (DHA) or a direct chain thereof Chain or branched C1-C6 alkyl esters, triglycerides or phospholipids thereof, alpha-linolenic acid (ALA) or straight-chain or branched C1-C6 alkyl esters thereof, triglycerides or phospholipids, stearidonic acid (stearidonic acid, SDA) or a straight or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, and eicosatetraenoic acid (ETA) or a straight or branched C1-C6 alkyl ester thereof , Any one or more selected from the group consisting of triglycerides or phospholipids thereof, the composition. According to claim 1, wherein the arthritis is senile arthritis, degenerative arthritis, autoimmune arthritis, osteoarthritis, rheumatoid arthritis, spondyloarthropathy, ankylosing spondylitis, psoriatic arthritis, gout, bacterial arthritis, juvenile rheumatoid arthritis, lupus, Behcet's disease, Reiter's syndrome, Pharmaceuticals for the prevention or treatment of any one or more arthritis selected from the group consisting of Lyme arthritis, adhesive capsulitis, frozen shoulder, tendon synovitis, elbow capsulitis, de Quervain's tendon synovitis, recurrent rheumatism, polymyalgia rheumatica and adult-type Still's disease enemy composition. The pharmaceutical composition for preventing or treating arthritis according to claim 1, which inhibits the expression of MCP-1, a chemokine that aggravates inflammation caused by arthritis. The pharmaceutical composition for preventing or treating arthritis according to claim 1, which reduces cartilage destruction by inhibiting the expression of cartilage-destroying factors, such as metalloproteinase 1 (MMP1), metalloproteinase 3 (MMP3), and metalloproteinase 13 (MMP13). The pharmaceutical composition for preventing or treating arthritis according to claim 1, comprising curcumin at a concentration of 2 mg/kg to 5000 mg/kg. The pharmaceutical composition for preventing or treating arthritis according to claim 1, wherein the omega-3 fatty acid is contained in a concentration of 2.2 mg/kg to 5500 mg/kg, and linoleic acid is contained in a concentration of 4.4 mg/kg to 11000 mg/kg. As a pharmaceutical composition for preventing or treating osteoporosis comprising curcumin, omega 3 fatty acids and linoleic acid as active ingredients,
The omega 3 fatty acid is eicosapentaenoic acid (EPA) or a linear or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, docosahexaenoic acid (DHA) or a direct chain thereof Chain or branched C1-C6 alkyl esters, triglycerides or phospholipids thereof, alpha-linolenic acid (ALA) or straight-chain or branched C1-C6 alkyl esters thereof, triglycerides or phospholipids, stearidonic acid (stearidonic acid, SDA) or a straight or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, and eicosatetraenoic acid (ETA) or a straight or branched C1-C6 alkyl ester thereof , Any one or more selected from the group consisting of triglycerides or phospholipids thereof, the composition. As a food composition for preventing or improving arthritis comprising curcumin, omega 3 fatty acids and linoleic acid as active ingredients,
The omega 3 fatty acid is eicosapentaenoic acid (EPA) or a linear or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, docosahexaenoic acid (DHA) or a direct chain thereof Chain or branched C1-C6 alkyl esters, triglycerides or phospholipids thereof, alpha-linolenic acid (ALA) or straight-chain or branched C1-C6 alkyl esters thereof, triglycerides or phospholipids, stearidonic acid (stearidonic acid, SDA) or a straight or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, and eicosatetraenoic acid (ETA) or a straight or branched C1-C6 alkyl ester thereof , Any one or more selected from the group consisting of triglycerides or phospholipids thereof, the composition. As a food composition for preventing or improving osteoporosis comprising curcumin, omega 3 fatty acids and linoleic acid as active ingredients,
The omega 3 fatty acid is eicosapentaenoic acid (EPA) or a linear or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, docosahexaenoic acid (DHA) or a direct chain thereof Chain or branched C1-C6 alkyl esters, triglycerides or phospholipids thereof, alpha-linolenic acid (ALA) or straight-chain or branched C1-C6 alkyl esters thereof, triglycerides or phospholipids, stearidonic acid (stearidonic acid, SDA) or a straight or branched C1-C6 alkyl ester thereof, a triglyceride or phospholipid thereof, and eicosatetraenoic acid (ETA) or a straight or branched C1-C6 alkyl ester thereof , Any one or more selected from the group consisting of triglycerides or phospholipids thereof, the composition.

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