JP2013001688A - Arthritis improvement composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an arthritis improvement composition capable of alleviating arthralgia in a short period of time, capable of retaining effects for a long period of time, and having both immediate effectivity and durability.SOLUTION: By ingesting the arthritis improvement composition combining glucosamines with carotenoids and/or eicosapentaenoic acid (EPA), the arthralgia can be alleviated in a short period of time and further the effects can be retained for a long period of time. Namely it is found that the arthritis improvement composition has arthritis improvement effects having both the immediate effectivity and durability, and the present invention is accomplished. The arthritis improvement composition obtained by the present invention may be powdered, granulated, tableted by tableting it, or filled in a soft capsule or a hard capsule. The arthritis improvement composition can be used in any states.

Description

  The present invention relates to a composition for improving arthritis which is expected to be used for "medicine", "food for specified health use", "so-called health food" and foods, beverages and confectionery.

Since joints are always mechanically stimulated by exercise, they are easily prone to inflammation. There are various causes of inflammation, such as joint deformation, bacterial / viral infection, trauma, immune diseases such as allergies, or metabolic abnormality due to nephritis, etc., but joint pain is often caused as a subjective symptom. As the population ages, the population suffering from arthritis such as knee and elbow pain is increasing, and pharmaceuticals and health foods that relieve such arthritis are on the market. This includes, for example, foods containing mucopolysaccharides such as chondroitin and glucosamine and / or amino sugars alone, or combinations of glucosamine and chondroitin as described in JP-T 9-503197 (Patent Document 1). Food.
JP-A-2000-53569 (Patent Document 2) discloses L-carnitines, glucosaminoglycans (such as chondroitin sulfate), glucosamine and excipients as compositions suitable for prevention and treatment of joint disorders. A composition containing is disclosed.

On the other hand, hyaluronic acid is a mucopolysaccharide and has been approved as a therapeutic drug (injection drug) for osteoarthritis of the knee. JP-A 2004-166616 (Patent Document 3) discloses a health food containing black vinegar, glucosamine, chondroitin sulfate, and hyaluronic acid as a health food that relieves symptoms such as joint pain. Has been.
In general, the effects of glucosamine, chondroitin sulfate, and hyaluronic acid are recognized to some extent, but they are slow-acting and cannot be experienced within about one month after ingestion of food. For this reason, many people would stop the intake of the food without having to experience these effects, there was a need to improve the slow-acting of food that have been used for such as a conventional joint pain. That is, there is a demand for pharmaceuticals, health foods and the like that can experience the effect of relieving joint pain in a short period of time and can maintain the effect for a long period of time.
Japanese National Patent Publication No. 9-503197 JP 2000-53569 A JP 2004-166616 A

  Therefore, an object of the present invention is to provide an arthritis improving composition that can relieve joint pain in a short period of time and can maintain an effect for a long period of time, and has both immediate effect and sustainability.

As a result of intensive studies, the present inventors can relieve joint pain in a short period of time by taking an arthritis improving composition that combines glucosamines and carotenoids and / or eicosapentaenoic acid (EPA). And found that the effect can be maintained for a long time. In other words, the arthritis improving composition of the present application was found to have an arthritis improving effect having both immediate effect and sustainability, which has not been heretofore, and the present invention was completed.
Therefore, this application includes the following invention.
(1) As an active ingredient for improving arthritis, glucosamines, carotenoids and / or eicosapentaenoic acid (EPA) are contained,
The composition for improving arthritis, wherein the carotenoid is one or a combination of two or more selected from the group of fucoxanthin, lutein, astaxanthin, zeaxanthin, β-carotene, α-carotene and β-cryptoxanthin. .

(2) An arthritis improving composition comprising glucosamines and fucoxanthin or glucosamines and fucoxanthin and eicosapentaenoic acid (EPA) as active ingredients for improving arthritis.

(3) The arthritis ameliorating composition contains interleukin-1β in knee synovial sac tissue
The composition for improving arthritis according to (2), wherein the composition has an anti-inflammatory action by suppressing (IL-1β) level and suppressing expression of cyclooxygenase-2 (COX-2).

(4) In a soft capsule formed by coating the capsule contents with a soft film,
The soft capsule film part consists of a film containing gelatin, a plasticizer and water,
The soft capsule content contains dietary fiber, a surfactant, and an active ingredient for improving arthritis,
The arthritis improving composition according to (3), which contains glucosamines, fucoxanthin and eicosapentaenoic acid (EPA) as an active ingredient for improving arthritis,
Features soft capsule.

(5) The arthritis improving composition contains dietary fiber and an active ingredient for improving arthritis,
A powder, granule, or tablet, which is an arthritis improving composition according to (3), which contains glucosamines and fucoxanthin as active ingredients for improving arthritis.

(6) The arthritis ameliorating composition according to (1), (2), (3), (4) or (5), characterized in that it is a food or drink with an indication that it is used for improving arthritis .

  By ingesting the arthritis improving composition of the present invention, joint pain can be relieved in a short period of time, and the effect can be maintained for a long period of time. That is, the present invention has an arthritis improving effect having both immediate effect and sustainability, which has not existed before.

FIG. 1 is a time-dependent change of the relative inflammation degree of the knee joint of each group when the inflammation degree of the knee joint of the model control group is 100%. FIG. 2 shows the results of measurement of interleukin-1β (IL-1β) level in the extract of knee synovial sac tissue. FIG. 3 shows the result of Western blotting analysis of cyclooxygenase-2 (COX-2) in knee synovial sac extract.

  As a result of extensive research, the present inventors have used glucosamines in combination with carotenoids and / or eicosapentaenoic acid (EPA) as active ingredients for improving arthritis in preparing an arthritis improving composition. As a result, it was found that joint pain can be relieved in a short period of time, and the effect can be maintained for a long period of time, and the present invention has been completed. The arthritis improving composition of the present invention contains glucosamines, carotenoids and / or eicosapentaenoic acid (EPA). Hereinafter, the principle of operation of the present invention and each of glucosamines, carotenoids, and eicosapentaenoic acid (EPA) of the present invention will be described.

<Operation principle of the present invention>
Interleukin-1β (IL-1β), which is involved in the principle of action of the present invention, is a pro-inflammatory cytokine secreted mainly by monocytes and macrophages. It has been reported that it is secreted from various cells such as cells, astrocytes, and microglia. In addition, IL-1β mediates inflammatory responses by inducing production of IL-2, IL-3, IL-6 and interferon in B cells, T cells, and NK cells, and is exposed to IL-1β. Endothelial cells and smooth muscle cells then synthesize prostaglandins and other arachidonic acid derivatives. In addition, IL-1β is found in the synovial fluid of arthritic patients, which causes osteoclast activation and degranulation of eosinophils and basophils.

Cyclooxygenase (COX), on the other hand, is involved in the production of prostaglandin G2, an intermediate in the process of producing prostaglandins (PG) and thromboxanes (TX) from arachidonic acid, an essential fatty acid in the body. It is an enzyme. In recent years, it has been clarified at the gene / protein level that there are two types of COX, cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2).
Of these, cyclooxygenase-2 (COX-2), which is involved in the principle of action of the present invention, is an enzyme that produces prostaglandins (PG), proteases, etc., and is also referred to as “inflammatory COX”. It is produced by sexual stimuli (macrophages, cytokines, endotoxins, etc.). That is, COX-2 is induced by inflammatory stimuli and causes an inflammatory response.

  The synovial sac (also known as synovial bursa) tissue involved in the working principle of the present invention is a flat vesicle between the muscles and tendons contacting each other or bone, cartilage, ligament, etc., and consists of synovium It is an organization. There is a small amount of synovial fluid in the cavity, reducing friction. There are many muscle attachments, especially in the vicinity of joints, and it is not easy to communicate with nearby joint cavities. The synovial sac includes the submuscular synovial sac, the subtendon synovial sac, the subfascial synovial sac between the fascia and the muscle and the tendon, and the bone and cartilage that are in contact with the skin and is related to the muscle There is no subcutaneous bursa.

  Although the principle of action of the present invention is not necessarily clear, in preparing an arthritis ameliorating composition, as an effective ingredient for ameliorating arthritis, glucosamines and carotenoids and / or eicosapentaenoic acid (EPA) should be taken orally. Suppresses the level of pro-inflammatory cytokine interleukin-1β (IL-1β) in knee synovial sac tissue earlier than when glucosamines are taken alone, and prostaglandins (PG) It is considered that the anti-inflammatory effect is exhibited with immediate effect by suppressing the expression of cyclooxygenase-2 (COX-2), which is an enzyme producing protease and protease, in the knee synovial sac tissue at an early stage.

  The glucosamines in the present invention are glucosamine or derivatives thereof or salts thereof. For example, glucosamine, glucosamine derivatives (N-acetylglucosamine, N-methyl-L-glucosamine, etc.), glucosamine salts (hydrochloride, sulfate, etc.) Or a combination of two or more kinds selected from the group of physiologically acceptable salts (eg, glucosamine hydrochloride, glucosamine sulfate, glucosamine phosphate, etc.). Among these, glucosamine, glucosamine hydrochloride, and N-acetylglucosamine are preferably used. In the present invention, the blending amount of glucosamines is suitably used at 10 to 90% by weight with respect to the whole arthritis improving composition. More preferably, it is 20 to 80%, and particularly preferably 30 to 70%.

  Glucosamine in the present invention is a kind of hexosamine and is a typical amino sugar. In nature, it is mainly D-form, 2-amino-2-deoxy-D-glucose, and is a constituent of polysaccharides of animals and plants and microorganisms, particularly mucopolysaccharides, glycoproteins and glycolipids. In the present invention, not only natural glucosamine but also synthetic products can be used, and commercially available glucosamine can be used.

  The glucosamine derivative in the present invention is preferably a compound that can be converted into glucosamine after being applied in vivo because glucosamine is a constituent of proteoglycan in vivo. In addition, as long as it is a compound which has the effect of this invention, it does not restrict in particular to them. Examples of the glucosamine derivative of the present invention include derivatives such as N-acetylglucosamine and N-methyl-L-glucosamine. Among these, N-acetylglucosamine, which is a compound that can be converted into glucosamine in the body, is preferable.

  As the salt of glucosamine or a derivative thereof in the present invention, a pharmacologically (pharmaceutically) or physiologically acceptable salt can be used. Examples of pharmacologically or physiologically acceptable salts include organic acid salts (for example, lactate, acetate, butyric acid, trifluoroacetate, fumarate, maleate, tartrate, citrate, succinate, etc. Acid salt, malonate, methanesulfonate, toluenesulfonate, tosylate, palmitic acid, stearic acid, etc., inorganic acid salt (eg hydrochloride, sulfate, nitrate, hydrobromide, phosphorus Acid salts) and the like. Preferred are glucosamine hydrochloride, glucosamine sulfate, and glucosamine phosphate.

  Carotenoids are yellow, red or purple polyene pigments derived from natural products and have an antioxidant function. The carotenoids that can be used in the present invention are one or a combination of two or more selected from the group of fucoxanthin, lutein, astaxanthin, zeaxanthin, β-carotene, α-carotene, and β-cryptoxanthin, Although it may be appropriately determined in consideration of the expected effect and the like, it is suitably used at 1 to 30% by weight with respect to the entire arthritis improving composition. More preferably, it is 3 to 20%, and particularly preferably 5 to 20%. Moreover, the composition ratio of the carotenoids in the carotenoids that can be used in the present invention varies depending on the manufacturer and the product, but the blended amount converted as the carotenoids is 0.01 to 20% by weight with respect to the total arthritis improving composition. Is preferably used. More preferably, it is 0.05 to 10%, and particularly preferably 0.06 to 5%. Among these, fucoxanthin is preferably used. Fucoxanthin is a reddish brown pigment component contained in trace amounts in brown algae such as kelp, hijiki, and seaweed, and any fucoxanthin sold in the market can be used in the present invention.

  Eicosapentaenoic acid (EPA) that can be used in the present invention is one of the essential fatty acids that are not synthesized in the body and is a kind of unsaturated fatty acids classified as omega-3 fatty acids. ) Is included in many. In the present invention, any purified fish oil containing eicosapentaenoic acid (EPA) or eicosapentaenoic acid (EPA) sold in the market can be used. The blending amount may be appropriately determined in consideration of the expected effect. For example, in the case of refined fish oil containing 28% as an EPA composition ratio, it is 1 to 60% in terms of weight percentage with respect to the entire arthritis improving composition. Preferably used. More preferably, it is 10 to 55%, and particularly preferably 20 to 50%. In addition, the composition ratio of EPA in the refined fish oil that can be used in the present invention varies depending on the manufacturer and product, but the blended amount converted as EPA is preferably 0.3 to 25% by weight with respect to the total arthritis improving composition. Used for. More preferably, it is 5 to 20%, and particularly preferably 10 to 15%.

  The arthritis improving composition obtained by the present invention can be used in any form, such as powder or granule, tableted tablet, or filled with soft capsule or hard capsule. Can do. In the present invention, glucosamines and carotenoids and / or eicosapentaenoic acid (EPA) can be used in combination as active ingredients for improving arthritis. In addition to these active ingredients for improving arthritis, powders, granules, tablets, soft capsules, In consideration of the dosage form such as hard capsules, surfactants, fats and oils, dietary fibers, formulation additives and the like can be appropriately used.

  For example, a combination of glucosamines and carotenoids and / or eicosapentaenoic acid (EPA) as an active ingredient for improving arthritis, and other arthritis improving compositions using surfactants (glycerin fatty acid esters, etc.), oils and fats, and dietary fibers as appropriate The object can be used as a soft capsule content. As the soft capsule film, in addition to a film containing gelatin, a plasticizer (such as glycerin), and water, a vegetable film containing starch, carrageenan, a plasticizer (such as glycerin), a metal salt, and water is also included in the present invention. Can be used.

  The surfactant that can be used in the present invention is one or a combination of two or more selected from the group of monoglycerin fatty acid ester, diglycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, and sorbitan fatty acid ester. The amount is suitably used at 1 to 30% by weight with respect to the total arthritis improving composition. More preferably, it is 1 to 20%, and particularly preferably 2 to 15%. Among these, monoglycerin fatty acid ester and diglycerin fatty acid ester are preferably used.

  Any oils and fats that can be used in the present invention can be used as long as they are edible oils and fats such as vegetable oils, animal oils, and processed oils. In the present specification, among vegetable oils and fats, those that are liquid at normal temperatures such as olive oil are referred to as “vegetable oils”, and those that are solid at normal temperatures such as coconut oil are referred to as “vegetable fats”. In the present specification, among animal fats and oils, those that are liquid at room temperature such as fish oil are referred to as “animal oil”, and those that are solid at room temperature such as beef tallow and lard are referred to as “animal fat”. Examples of the processed fats and oils include hydrogenated processed fats and oils that are hydrogenated to vegetable oils and animal oils, or MCT (medium chain fatty acid triglyceride).

As oils and fats of the present invention, arthritis is improved by combining liquid oils (liquid at room temperature) such as vegetable oils and animal oils and solid oils (solid at room temperature) such as vegetable fats, animal fats and hydrogenated oils and fats. The viscosity and physical properties of the composition can be freely adjusted in accordance with the application. When the arthritis improving composition is used as the soft capsule content, it is desirable to adjust the viscosity of the arthritis improving composition to 5000 to 100000 Pa · s, but 20 to 50% liquid oil (liquid at room temperature) and 2 to 20% combination of solid fat, such as hydrogenated working oil (solid at room temperature) can be mentioned as an example of a case of using the arthritis improving composition as soft contents.
The blending amount of the oil or fat of the present invention is suitably used at 1 to 80% by weight with respect to the arthritis improving composition. More preferably, it is 10 to 70%, and particularly preferably 30 to 60%.

  The liquid oil (liquid at room temperature) that can be used in the present invention includes corn oil, soybean oil, salad oil, sesame oil, rapeseed oil, rice bran oil, koji oil, koji oil, safflower oil, palm oil, cottonseed oil, sunflower oil, olive Oil, peanut oil, almond oil, avocado oil, hazelnut oil, walnut oil, grape seed oil, mustard oil, lettuce oil, perilla oil, linseed oil, castor oil, paulownia oil, etc., or whale oil, coconut oil, liver oil One or two or more combinations selected from a group of fish oils such as, for example, can be mentioned as examples, but are not limited thereto. The blending amount of the liquid oil (liquid at room temperature) that can be used in the present invention may be appropriately determined in consideration of the expected effects and the like, but is preferably used at 1 to 80% by weight with respect to the arthritis improving composition. It is done. More preferably, it is 10 to 70%, and particularly preferably 40 to 60%.

  Solid fats (solid at room temperature) that can be used in the present invention include hydrogenated processed fats and oils such as plant-derived hydrogenated processed fats and fats, animal-derived hydrogenated processed fats and oils, vegetable fats such as cocoa butter and palm oil, lard Examples include 1 type or 2 or more types of combinations selected from the group consisting of (tallow fat), het (beef tallow), chicken fat, rosin, sheep fat, horse fat, milk fat and other animal fats. It is not limited to these. The amount of solid fat (solid at room temperature) that can be used in the present invention may be appropriately determined in consideration of the expected effect, the dosage form of the arthritis improving composition, the physical properties according to the application, etc. It is preferably used in an amount of 1 to 20% by weight based on the product. More preferably, it is 1 to 15%, and particularly preferably 2 to 10%. Examples of the plant-derived hydrogenated processed fats and oils in the present invention include hardened oil, margarine, shortening oil, and the like, and edible fats and oils that are either hydrogenated or partially hydrogenated can be used. Are preferably used. Moreover, lard etc. are mention | raise | lifted as an animal-derived hydrogenated processed oil and fat in this invention, What hydrogenated or partially hydrogenated edible oil and fat can be used.

In the present invention, glucosamines and carotenoids and / or eicosapentaenoic acid (EPA) can be used in combination as active ingredients for improving arthritis. In addition to these active ingredients for improving arthritis, powders, granules, tablets, soft capsules, Dietary fiber can be appropriately used in consideration of dosage forms such as hard capsules. By blending dietary fiber, the active ingredient for improving arthritis can be transferred into the blood efficiently and continuously.
The dietary fiber that can be used in the present invention is water-soluble dietary fiber, or insoluble dietary fiber, or a combination of water-soluble dietary fiber and insoluble dietary fiber. The blending amount of dietary fiber may be appropriately determined in consideration of the expected effect and the like, but is preferably used at 1 to 25% by weight with respect to the arthritis improving composition. More preferably, it is 1 to 20%, and particularly preferably 2 to 10%.

  Water-soluble dietary fiber that can be used in the present invention is resistant to dextrin, polydextrose, alginic acid, low molecular weight sodium alginate, galactomannan, glucomannan, water-soluble pectin, fucoidan, guar gum, guar gum degradation product, water-soluble lignin, chondroitin One type or a combination of two or more types selected from the group. Among these, indigestible dextrin and polydextrose are preferably used. The insoluble dietary fiber that can be used in the present invention is one or a combination of two or more selected from the group consisting of cellulose, bran, hemicellulose, insoluble pectin, insoluble lignin, glucan, inulin, carrageenan, agarose, chitin, and chitosan. .

  In the present invention, formulation additives such as powders, granules, tablets, soft capsules, hard capsules and the like can be used as needed. Examples of formulation additives used in powders, granules, and tablets include lubricants, saccharides, binders, excipients, fluidizers, disintegrants, and the like. The lubricant is not particularly limited as long as it can be used in foods. For example, stearic acid, stearic acid metal salts such as magnesium stearate and calcium stearate, sucrose fatty acid ester or glycerin fatty acid ester, and hardened fats and oils. Etc. The saccharide is not particularly limited as long as it can be used for foods, and examples thereof include monosaccharides, disaccharides, sugar alcohols, oligosaccharides and the like. Examples of monosaccharides include glucose, xylose, galactose, and fructose. Examples of the disaccharide include trehalose, sucrose, lactose, palatinose and the like. Examples of the sugar alcohol include maltitol, erythritol, sorbitol, xylitol and the like. Examples of the oligosaccharide include raffinose, inulooligosaccharide (chicory oligosaccharide), palatinose oligosaccharide and the like. The binder is not particularly limited as long as it can be used for foods and the like, and examples thereof include gelatin and pullulan. The excipient is not particularly limited as long as it can be used in foods and the like, and examples thereof include dextrin, crystalline cellulose, low-substituted hydroxypropylcellulose and the like. The fluidizing agent is not particularly limited as long as it can be used in foods and the like, and examples thereof include calcium phosphate, calcium hydrogen phosphate, and fine silicon dioxide. The disintegrant is not particularly limited as long as it can be used in foods and the like, and examples thereof include corn starch, potato starch, and partially pregelatinized starch.

  In the present invention, glucosamines and carotenoids and / or eicosapentaenoic acid (EPA) can be used in combination as active ingredients for improving arthritis, but from other groups of hydrophilic functional components and lipophilic functional components. Even if one or two or more selected combinations are contained, the effects of the present invention are not inhibited.

  Examples of hydrophilic functional ingredients that can be used in the present invention include water-soluble vitamins composed of vitamin B group (vitamin B1, vitamin B2, niacin, vitamin B6, vitamin B12, pantothenic acid, folic acid, biotin) and vitamin C, rutin, Taurine, rice vinegar, rice black vinegar, brown rice vinegar, brown rice black vinegar, barley black vinegar, pearl barley vinegar, wheat vinegar, apple vinegar, grape vinegar, koji vinegar, plum vinegar, tomato vinegar, moromi vinegar, water soluble oligosaccharide, water soluble Dextrin, isoflavone, flavonol, flavanone, anthocyanin, flavanol, flavone, catechin, tannin, flavonoid, chlorogenic acid, phenylcarboxylic acid, ellagic acid, lignan, curcumin, coumarin, alpha lipoic acid, xylitol, soy peptide, soy isoflavone, citric acid Glycine, alanine, serine, cysteine, cystine, methi Examples include, but are not limited to, onine, glutamine, arginine, histidine, collagen peptide, and any hydrophilic functional component can be used.

  The blending amount of the glucosamines of the composition for improving arthritis in the present invention and the hydrophilic functional component other than carotenoids and / or eicosapentaenoic acid (EPA) may be appropriately determined in consideration of the expected effects and the like. It is preferably used in an amount of 1 to 50% by weight based on the entire improved composition. More preferably, it is 5 to 40%, and particularly preferably 10 to 30%.

  Examples of lipophilic functional ingredients that can be used in the present invention include fat-soluble vitamins (vitamin A, vitamin D, vitamin E, vitamin K), coenzyme Q10, DHA (docosahexaenoic acid), lycopene, plant sterols, linoleic acid, linolenic acid, Examples thereof include, but are not limited to, arachidonic acid, oleic acid, and squalene, and any and all lipophilic functional ingredients can be used.

  The blending amount of the glucosamines of the composition for improving arthritis in the present invention and the lipophilic functional component other than carotenoids and / or eicosapentaenoic acid (EPA) may be appropriately determined in consideration of the expected effect and the like. It is preferably used in an amount of 1 to 50% by weight with respect to the entire absorption promoting composition. More preferably, it is 5 to 40%, and particularly preferably 10 to 30%.

  The present invention will be described in more detail by the following examples, but the present invention is not limited to these examples. Prior to the examples, the test methods used in the examples will be described.

<Adjustment of experimental sample>
In this example, using a rat knee arthritis model induced by kaolin and carrageenan, eicosapentaenoic acid contained in glucosamine hydrochloride (GlucosamineHCl: GAH), fucoxanthin (FUCO) and / or refined fish oil (EPA28%) (eicosapentaenoic acid: EPA) alone and six (present invention product 2 kinds, Comparative example 4 kinds) of the combination of the studied arthritis improvement by composition. Specifically, sample compositions of the present invention product 1, the present invention product 2, the comparative example 1, the comparative example 2, the comparative example 3, and the comparative example 4 were prepared with the component compositions shown in Table 1.
The product 1 of the present invention is a composite composition of three active ingredients containing glucosamine hydrochloride, fucoxanthin and EPA, and the product 2 of the present invention is a composite composition of two active ingredients containing glucosamine hydrochloride and fucoxanthin.
On the other hand, Comparative Example 1 is a two-component active ingredient composite composition containing glucosamine hydrochloride and EPA. Comparative Example 2 is an active ingredient 1 type composition containing glucosamine hydrochloride, Comparative Example 3 is an active ingredient 1 type composition containing fucoxanthin, and Comparative Example 4 is an active ingredient 1 type composition containing EPA. It is a thing.

  In addition, since the fucoxanthin used in the present example is 1% Powder of Fucoxanthin (manufactured by Beijing Kinka Group), for example, the blending amount of Fucoxanthin 1% Powder of Product 1 of the present invention is 6.45%. However, the compounding amount converted as fucoxanthin is 0.0645%. In addition, since the purified fish oil EPA-28 (manufactured by Nippon Suisan Co., Ltd.) used in this example has a composition ratio of EPA of 28%, for example, the purified fish oil EPA-28 (EPA 28%) of the product 1 of the present invention. Is 48.39%, but the amount converted as EPA is 13.55% (48.39% × 0.28 = 13.55%).

Invention product 1
In the composition according to the present invention product 1 in Table 1, glycerol fatty acid ester is added to EPA 28% as a primary stirring treatment, and the rotational speed is gradually increased using a homojetter (high speed stirrer) at a temperature of 65 ± 5 ° C., 4000 rpm And homogenized for 20 minutes. Then, the homogenized intermediate composition is cooled to a temperature of 40 ± 10 ° C., and as a secondary stirring process, glucosamine hydrochloride, fucoxanthin 1%, dietary fiber (digestible dextrin) are added to the homogenized intermediate composition. In addition, by gradually increasing the number of rotations using a homojetter (high speed stirrer) at a temperature of 40 ± 10 ° C. and stirring the mixture at 4000 rpm for 20 minutes, the glucosamine hydrochloride and fucoxanthin of the product 1 of the present invention are obtained. A composite composition of three active ingredients containing EPA was obtained. The triple complex composition was used by oral gavage.
As the glycerin fatty acid ester, two types of HLB 7.4 (trade name: Poem DO-100V) manufactured by Riken Vitamin Co., Ltd. and HLB 3.8 (trade name: Excel S-95) manufactured by Kao Corporation were used.

Invention product 2
In the composition according to the present invention product 2 in Table 1, as the primary stirring treatment, glycerin fatty acid ester is added to MCT, and the number of revolutions is gradually increased using a homojetter (high speed stirrer) at a temperature of 65 ± 5 ° C. at 4000 rpm. Stir for 20 minutes to homogenize. Then, the homogenized intermediate composition is cooled to a temperature of 40 ± 10 ° C., and as a secondary stirring process, glucosamine hydrochloride, fucoxanthin 1%, dietary fiber (digestible dextrin) are added to the homogenized intermediate composition. In addition, by gradually increasing the number of revolutions using a homojetter (high-speed stirrer) at a temperature of 40 ± 10 ° C. and stirring for 20 minutes at 4000 rpm, the product is mixed with glucosamine hydrochloride and fucoxanthin of the product 2 of the present invention. Two active ingredient composite compositions were obtained. The two complex compositions were used by oral gavage.
As the glycerin fatty acid ester, two types of HLB 7.4 (trade name: Poem DO-100V) manufactured by Riken Vitamin Co., Ltd. and HLB 3.8 (trade name: Excel S-95) manufactured by Kao Corporation were used.

Comparative Example 1
In the composition according to Comparative Example 1 in Table 1, as a primary stirring treatment, glycerin fatty acid ester is added to 28% of EPA, and the number of revolutions is gradually increased using a homojetter (high speed stirrer) at a temperature of 65 ± 5 ° C. at 4000 rpm. Stir for 20 minutes to homogenize. Then, the homogenized intermediate composition is cooled to a temperature of 40 ± 10 ° C., and as a secondary stirring treatment, glucosamine hydrochloride and dietary fiber (digestible dextrin) are added to the homogenized intermediate composition to obtain a temperature of 40 By gradually increasing the number of rotations using a homojetter (high speed stirrer) at ± 10 ° C. and stirring the mixture at 4000 rpm for 20 minutes to achieve homogenization, a composite composition of glucosamine hydrochloride of Comparative Example 1 and EPA containing two active ingredients is combined. I got a thing. The two complex compositions were used by oral gavage.
As the glycerin fatty acid ester, two types of HLB 7.4 (trade name: Poem DO-100V) manufactured by Riken Vitamin Co., Ltd. and HLB 3.8 (trade name: Excel S-95) manufactured by Kao Corporation were used.

Comparative Example 2
In the composition according to Comparative Example 2 in Table 1, glycerin fatty acid ester is added to MCT as a primary stirring treatment, and the number of revolutions is gradually increased using a homojetter (high speed stirrer) at a temperature of 65 ± 5 ° C., and 20 at 4000 rpm. Stir for minutes to homogenize. Then, the homogenized intermediate composition is cooled to a temperature of 40 ± 10 ° C., and as a secondary stirring treatment, glucosamine hydrochloride and dietary fiber (digestible dextrin) are added to the homogenized intermediate composition to obtain a temperature of 40 By gradually increasing the number of revolutions using a homojetter (high speed stirrer) at ± 10 ° C. and stirring the mixture at 4000 rpm for 20 minutes, the composition of active ingredient 1 type containing the glucosamine hydrochloride of Comparative Example 2 is obtained. It was. The active ingredient 1 type composition was used by oral gavage.
As the glycerin fatty acid ester, two types of HLB 7.4 (trade name: Poem DO-100V) manufactured by Riken Vitamin Co., Ltd. and HLB 3.8 (trade name: Excel S-95) manufactured by Kao Corporation were used.

Comparative Example 3
In the composition according to Comparative Example 3 in Table 1, glycerin fatty acid ester was added to MCT as a primary stirring treatment, and the number of revolutions was gradually increased using a homojetter (high-speed stirrer) at a temperature of 65 ± 5 ° C. and 20 at 4000 rpm. Stir for minutes to homogenize. Then, the homogenized intermediate composition is cooled to a temperature of 40 ± 10 ° C., and as a secondary stirring treatment, fucoxanthin 1% and dietary fiber (digestible dextrin) are added to the homogenized intermediate composition, and the temperature is increased. By gradually increasing the number of rotations using a homojetter (high speed stirrer) at 40 ± 10 ° C. and stirring for 20 minutes at 4000 rpm, the composition of fucoxanthin blended in Comparative Example 3 is obtained. It was. The active ingredient 1 type composition was used by oral gavage.
As the glycerin fatty acid ester, two types of HLB 7.4 (trade name: Poem DO-100V) manufactured by Riken Vitamin Co., Ltd. and HLB 3.8 (trade name: Excel S-95) manufactured by Kao Corporation were used.

Comparative Example 4
In the composition according to Comparative Example 4 in Table 1, as the primary stirring treatment, glycerin fatty acid ester is added to 28% EPA and MCT, and the rotational speed is gradually increased using a homojetter (high speed stirrer) at a temperature of 65 ± 5 ° C. The mixture was homogenized by stirring at 4000 rpm for 20 minutes. Then, the homogenized intermediate composition is cooled to a temperature of 40 ± 10 ° C., and as a secondary stirring treatment, dietary fiber (indigestible dextrin) is added to the homogenized intermediate composition at a temperature of 40 ± 10 ° C. The number of rotations was gradually increased using a homojetter (high-speed stirrer), and the mixture was stirred and homogenized at 4000 rpm for 20 minutes. The active ingredient 1 type composition was used by oral gavage.
As the glycerin fatty acid ester, two types of HLB 7.4 (trade name: Poem DO-100V) manufactured by Riken Vitamin Co., Ltd. and HLB 3.8 (trade name: Excel S-95) manufactured by Kao Corporation were used.

<Animal group and sample administration method>
The animal group used in this example is a specific pathogen-free (SPF) male Sprague-Dawley rat that is not infected with a specific pathogen, 5 weeks old, and has a body weight of 168 to 226 g. Received from the University Test Animal Center. Rats were quarantined and acclimatized for 7 days under the experimental conditions in the animal laboratory, and at the same time the health status was observed. The number of rats was 5 for each gauge, and soft sterilized wood chips were laid in the gauge. The animal laboratory temperature was maintained at 23-25 ° C. and relative humidity at 50-60%. Ventilation frequency was about 12 times / hour, lighting was 12 hours / day, and darkness was 12 hours / day. The animal feed was a rod-shaped dry basic animal feed for laboratory animals, and the drinking water was boiled and cooled, and tap water was given for free consumption.

Rats were grouped randomly by body weight, respectively, the inventive product group 1, the present invention product group 2, Comparative Example group 1, Comparative Example group 2, Comparative Example group 3, Comparative Example Group 4, control group, model control Groups were divided into 8 groups of 10 animals per group. Distilled water in the control group and the model control group, the other 6 groups, the present invention product 1 is subject compositions, the present invention product 2, Comparative Example 1, Comparative Example 2, Comparative Example 3, respectively orally administered Comparative Example 4 did.
Inventive product 1 and invented product 2, which are test compositions, were used in an amount of 3.1 g / kg body weight (glucosamine hydrochloride 1000 mg / kg body weight, fucoxanthin 1% 200 mg / kg body weight, EPA28% 1500 mg / kg body weight), and product 2 of the present invention is 3.1 g / kg body weight (glucosamine hydrochloride 1000 mg / kg body weight, fucoxanthin 1% 200 mg / kg body weight). On the other hand, the amount used in Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4, which are test compositions, was 2.9 g / kg body weight (glucosamine hydrochloride 1000 mg / kg body weight, EPA 28% 1500 mg, respectively). / kg body weight), Comparative Example 2 is 2.9 g / kg body weight (glucosamine hydrochloride 1000 mg / kg body weight), and Comparative Example 3 is 2.1 g / kg body weight (fucoxanthin 1% 200 mg / kg body weight) Comparative Example 4 is 2.9 g / kg body weight (EPA 28% 1500 mg / kg body weight).

<Method of developing arthritis>
On day 0, distilled water in the control group and the model control group, the other 6 groups, the present invention product 1 is subject compositions, the present invention product 2, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example Each of 4 was orally administered to rats. Its administration to one hour later, except the control group, model control group and the test composition group (present invention product group 1, the present invention product group 2, Comparative Example group 1, Comparative Example group 2, Comparative Example group 3, Comparative Example Group Arthritis was developed in the rats of 4) using kaolin (manufactured by Sigma) and carrageenan (manufactured by Sigma).
As a specific method for the development of arthritis, rats were anesthetized with pentobarbital sodium (30 mg / kg body weight) by intraperitoneal injection. The solution was injected with 0.1 ml of 4% kaolin and 2% λ-carrageenan suspension prepared with sterile physiological saline. From the next day, the test composition was orally administered once a day for 6 consecutive days.

<Knee joint inflammation evaluation method>
<Knee joint inflammation>
On days 1, 3, 5, and 7 when arthritis developed, the diameter (cm) of the left and right knee joints of each group of rats (10 rats) was measured with a digital caliper in the horizontal direction. The degree of inflammation (%) of the knee joint was calculated using the following formula.
Knee joint inflammation (%) = [Right knee joint diameter (cm)-Left knee joint diameter (cm)] / Right knee joint diameter (cm) x 100
<Relative inflammation of knee joint>
The ratio of the knee joint inflammation degree of each group when the knee joint inflammation degree of the model control group was 100% was calculated as the relative inflammation degree of the knee joint.

<Experimental evaluation results>
Table 2 shows the results of investigation and analysis of the degree of inflammation of the knee joint and the relative inflammation of the knee joint 1, 3, 5 and 7 days after the induction of knee arthritis on the right side of the rat with kaolin and carrageenan suspension. In addition, FIG. 1 shows changes with time in the relative inflammation level of the knee joints of each group when the knee joint inflammation level of the model control group is 100%.

From Table 2 and FIG. 1, the effect of suppressing the degree of inflammation of knee joints of the products 1 and 2 of the present invention is immediate effect as compared with the model control group and Comparative Example 1, Comparative Example 2, Comparative Example 3, and Comparative Example 4. Both sustainability was excellent.
In particular, the relative inflammation degree of the product 1 of the present invention containing glucosamine hydrochloride, fucoxanthin and EPA is suppressed to 43.8% from the first day compared to the model control group, and 32.5% on the seventh day. % Anti-inflammatory action was recognized, and both immediate effect and persistence were very excellent (FIG. 1).
In addition, the relative inflammation degree of the product 2 of the present invention containing glucosamine hydrochloride and fucoxanthin is suppressed to 68.6% from the first day compared to the model control group, and to 37.3% on the seventh day. The anti-inflammatory action was recognized up to the present, and after the product 1 of the present invention, both immediate effect and sustainability were excellent.

Next, the mechanism of the present invention, which can relieve joint pain in a short period of time and can maintain the effect for a long period of time, has both immediate effect and sustainability, was examined.
<Adjustment of experimental sample>
The present invention product 1 in Table 1 were prepared as in Example 1, the present invention product 2, Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Example 2, respectively present invention product 1 ', The product of the present invention 2 ′, Comparative Example 1 ′, Comparative Example 2 ′, Comparative Example 3 ′, and Comparative Example 4 ′ were used.

<Animal group and sample administration method>
The animal group used in this Example 2 is a male Sprague-Dawley rat of specific pathogen-free (SPF) that is not infected with a specific pathogen, 6 weeks old, and has a body weight of 208 to 242 g. Was the same as in Example 1.

<Method of developing arthritis>
On day 0, distilled water in the control group and the model control group, the other 6 groups, the present inventions are subject composition 1 ', the present invention product 2', Comparative Example 1 'and Comparative Example 2' and Comparative Example 3 ′ and Comparative Example 4 ′ were orally administered to rats. Its administration to one hour later, except the control group, model control group and the test composition group (present invention product group 1 ', the present invention product group 2', Comparative Example group 1 'and Comparative Example group 2', Comparative Example Group 3 ', Rats of Comparative Example Group 4') developed arthritis using kaolin (manufactured by Sigma) and carrageenan (manufactured by Sigma).
As a specific method for the development of arthritis, rats were anesthetized with pentobarbital sodium (30 mg / kg body weight) by intraperitoneal injection. The solution was injected with 0.1 ml of 4% kaolin and 2% λ-carrageenan suspension prepared with sterile physiological saline. The test composition was orally administered by gavage again 8 hours after the injection of 0.1 ml of 4% kaolin and 2% λ-carrageenan suspension.

<Evaluation method>
<Preparation of knee synovial sac tissue extract>
The preparation of knee synovial sac tissue extract was prepared by Method 1 (Penido C, Conte FP, Chagas MSS, Rodrigues CAB, Pereira JFG.
Antiinflammatory
effects of natural tetranortriterpenoids isolated
from Carapa guianensis. Aublet on zymosan
induced arthritis in mice. Inflammres 2006; 55: 457-464)
2 (Jean YH, Chen WF, Duh CY, Huang
SY, Hsu CH, Lin CS, Sung CS, Chen IM, Wen ZH.
Inducible nitric oxide synthase cyclooxygenase-2
participate in anti-inflammatory and analgesi effects
of the natural marine compound lemnalol from Formosa
soft coral Lemnalia cervicorni. Eur J pharm 2008; 578: 323-331.)
As a reference, I changed it a little.

  24 hours after injection of 0.1 ml of 4% kaolin and 2% λ-carrageenan suspension (25 hours after gavage of the first test composition: about 1 day later) After rats were anesthetized with ether and killed, the right hind paw was cut off and washed with ice-cold phosphate-buffered saline (PBS). Then, the knee synovial sac tissue is taken out and placed in a glass homogenizer together with ice-cold extraction buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 1% Triton X-100), and then mixed protein enzyme inhibitor (protease inhibitor cocktail ) Was added. After manual polishing in an ice bath, the mixture was transferred to a small centrifuge and centrifuged (20,000 × g, 10 min, 4 ° C.), and the resulting supernatant was stored frozen (−70 ° C.). The amount of protein in the supernatant solution was measured with a Bio-Rad protein measurement reagent kit.

<Enzyme immunosorbent assay>
Of HizaNamera sacs tissue extracts were measured interleukin -1β (IL-1β) levels are proinflammatory cytokines (pro-inflammatory cytokines). The enzyme immunosorbent measurement (ELISA) test was adopted as the measurement method. Corresponding antibody pairs were purchased from R & D Systems and operated according to the instructions for use.

<Western blotting analysis of cyclooxygenase-2 (COX-2)>
Western blotting (Western Blotting) analysis Jean et al and Rosengren Rayori introduction methods 1 (Jean YH, Chen WF, Duh CY, Huang
SY, Hsu CH, Lin CS, Sung CS, Chen IM, Wen ZH.
Inducible nitric oxide synthase cyclooxygenase-2
participate in anti-inflammatory and analgesi effects
of the natural marine compound lemnalol from Formosa
soft coral Lemnalia cervicorni. Eur J pharm 2008; 578: 323-331.)
2 (Rosengren S, Firestein GS, Boyle DL. Measurement of
inflammatory biomarkers in synovial tissue extracts by enzyme-linked immunosorbent assay. Clin Diagn Lab Immunol; 2003: 1002-1010.)
Adopted, but changed a little.

Equal volume of sample buffer (2% SDS, 10% glycerol, 0.1% bromophenol blue,
2% -2-mercaptoethanol, 50 mM Tris-HCl, pH 7.2) was added to the knee synovial sac tissue extract. Make SDS-polyacrylamide (SDS-PAGE), perform general electrophoresis (150V, 90 minutes), and then protein on polyvinylidene difluoride (PVDF) membrane (Immobilon-P, Millipore, 0.45) The membrane was sealed with Tris-buffered saline (Tris-buffered saline, TBS) containing 5% nonfat milk at room temperature for 1 hour. The membrane was incubated in TBS containing anti-COX-2 antibody (1: 1000 dilution, Cayman, Chemical, USA) for 180 minutes at room temperature. After washing 3 times (15 minutes) in TBS containing 0.1% Tween-20, goat IgG (anti-mouse or anti-rabbit) labeled with horseradish peroxidase as a membrane and secondary antibody (1: 2000 dilution) , Santa, Cruz Biotechnology, USA) was incubated at room temperature for 60 minutes.
Enhanced chemiluminescence (ECL kit, Santa Cruz Biotechnology, USA) was used for color development, and immunoreactive protein levels were taken using densitometer measurements.

<Experimental evaluation results>
The results of measuring interleukin-1β (IL-1β) levels in the knee synovial sac extract are shown in FIG. Further, cyclooxygenase HizaNamera sacs tissue extracts -2: 3 a (Cyclooxygenase-2 COX-2) Western blotting analysis of.

From FIG. 2, in the situation where only 25 hours (about 1 day) have passed since the first oral gavage administration of the test composition, the interleukin-1β (IL-1β) level was 14.39 for the product 1 ′ of the present invention. the present invention product 2 'is 16.90, the IL-l [beta] levels 36.20 model control group compared to the time taken as 100%, the present invention product 1' is 39.8%, the present invention product 2 ' A reduction to 46.7% was observed.
Similarly, even when the IL-1β level 29.36 of Comparative Example 2 ′ (GAH) using glucosamine hydrochloride alone was set as 100%, the product 1 ′ of the present invention was 49.0%, and the product 2 ′ of the present invention 2 ′. Was reduced to 57.6%. That is, rather than using glucosamine hydrochloride alone as in Comparative Example 2 ′ (GAH), by using fucoxanthin and EPA in combination as in the present product 1 ′ and the present product 2 ′, It was confirmed that glucosamine hydrochloride, which had a problem in immediate effect, remarkably enhanced the arthritis improving effect.

From FIG. 3, cyclooxygenase-2 (COX-2) protein expression in knee synovial sac tissue extract in a situation where only 25 hours (about 1 day) have passed since the first oral gavage of the test composition the amount (relative concentration), the product of the present invention 1 'is 62.65, the present invention product 2' is 54.54, and COX-2 protein expression level of the model control group (relative concentration) 181.10 100% As compared with the above, it was confirmed that the product 1 'of the present invention was reduced to 34.6% and the product 2' of the present invention was reduced to 30.1%.
Similarly, even when the COX-2 protein expression level (relative concentration) 179.35 of Comparative Example 2 ′ (GAH) using glucosamine hydrochloride alone is 100%, the product 1 ′ of the present invention is 34.9%. The product 2 'of the present invention was reduced to 30.4%. That is, rather than using glucosamine hydrochloride alone as in Comparative Example 2 ′ (GAH), by using fucoxanthin and EPA in combination as in the present product 1 ′ and the present product 2 ′, It was confirmed that glucosamine hydrochloride, which had a problem in immediate effect, remarkably enhanced the arthritis improving effect.

  In Example 2, distilled water or a test composition was forcibly administered orally on day 0, and one hour later, a model control group and a test composition group (invention product group 1 ′, the present invention), excluding the control group. Article group 2 ', comparative example group 1', comparative example group 2 ', comparative example group 3', comparative example group 4 ') were injected with an arthritis-inducing agent and tested again 8 hours after the injection. The composition was administered by oral gavage. Then, in a situation where only a very short time (about 1 day) has passed since 16 hours (25 hours after the first oral gavage administration of the test composition, ie, 24 hours after the injection), glucosamine hydrochloride Of the present invention 1 'containing glucosamine hydrochloride and fucoxanthin by forced oral administration of cyclooxygenase-2 (COX- It was found that the expression of 2) was clearly suppressed and IL-1β level was clearly reduced.

  Moreover, in Example 1, compared with the model control group and other Comparative Examples 1 to 4, the products 1 and 2 of the present invention exhibit excellent immediate effects from the first day, and their anti-inflammatory action Has been shown to be lasting until day 7. That is, the anti-inflammatory action of fucoxanthin and EPA remarkably enhances the arthritis improving effect of glucosamine hydrochloride, and has the effect of improving arthritis having both immediate effect and sustainability (Example 1 and Example). It became clear by 2). This is a fact first found by the present inventors.

  According to the Japanese Pharmacopoeia, General Rules for Tablets “Tablets”, tablets (300 mg / tablet, dosage form 9 mmФ) were produced using the tablet formulation shown in Table 3 below.

  The tablet of Example 3 can relieve arthralgia in a short period of time and can maintain the effect for a long period of time, similar to the product 2 of the present invention 2 of Example 1 and the product 2 ′ of the present invention of Example 2. It is an arthritis improving composition that has both immediate effect and durability.

  The present invention can be used in the fields of “pharmaceuticals”, “special health foods”, “so-called health foods”, “functional foods” and foods, beverages and confectionery.

Claims (6)

Contains glucosamines, carotenoids and / or eicosapentaenoic acid (EPA) as an active ingredient for improving arthritis,
The composition for improving arthritis, wherein the carotenoid is one or a combination of two or more selected from the group of fucoxanthin, lutein, astaxanthin, zeaxanthin, β-carotene, α-carotene and β-cryptoxanthin. .   An arthritis improving composition containing glucosamines and fucoxanthin, or glucosamines, fucoxanthin and eicosapentaenoic acid (EPA) as active ingredients for improving arthritis.   The composition for improving arthritis has an anti-inflammatory effect by suppressing interleukin-1β (IL-1β) level in knee synovial sac tissue and suppressing expression of cyclooxygenase-2 (COX-2) The arthritis improving composition according to claim 2, wherein the composition is an arthritis improving composition. In the soft capsule formed by coating the capsule contents with a soft film,
The soft capsule film part consists of a film containing gelatin, a plasticizer and water,
The soft capsule content contains dietary fiber, a surfactant, and an active ingredient for improving arthritis,
The arthritis ameliorating composition according to claim 3, comprising glucosamines, fucoxanthin and eicosapentaenoic acid (EPA) as active ingredients for improving arthritis.
Features soft capsule. The arthritis improving composition contains dietary fiber and an active ingredient for improving arthritis,
A powder, granule, or tablet, which is an arthritis improving composition according to claim 3, containing glucosamines and fucoxanthin as active ingredients for improving arthritis.   The arthritis improving composition according to claim 1, 2, 3, 4, or 5, wherein the composition is a food or drink with an indication that it is used for improving arthritis.