KR100830553B1 - Inflammation-related diseases prevention and treatment compositions - Google Patents

Abstract

The present invention is independent ( Aralia) cordata THUNB.) And Horseback Riding ( Cimicifuga) The present invention relates to a composition containing a mixed herbal extract of heracleifolia Kom.), and in particular, the composition of the present invention exhibits cartilage tissue protection effect, anti-inflammatory and analgesic effect, and thus may be used as a composition for preventing and treating inflammation-related diseases, especially arthritis. Can be.

Description

Composition for the prevention and treatment of inflammation-related diseases, including poisonous and horse-riding mixed herbal extracts {Composition comprising the mixed extract of Aralia cordata THUNB. and Cimicifuga heracleifolia Kom. for preventing and treating inflammatory disease}

The present invention relates to a composition for the treatment of inflammation-related diseases containing the extracts of poisonous and equestrian mixed herbs as an active ingredient.

In oriental medicine, arthritis is mainly expressed as nausea, and the clinical manifestation of nausea is mainly pain, and pain can be caused by pathological problems of blood circulation. The main external factors can be classified into wind, cold, damp, and heat.These causes the blood circulation problems such as joints and muscles, and pains. Limitations of range of motion appear. Internal factors are the major causes of the liver and kidneys, the muscles belonging to the liver, the liver presides over blood, and this is the muscle joint To nourish (滋養) to regulate the blood of the liver should be smooth to prevent pathological products such as blood (瘀血). In addition, God manages the bone (腎 主 腎) and regulates the human body (精), this tablet (精) functions to nourish the bone marrow and to limnate the skeleton. Therefore, the abnormalities of the joints are closely related to the functions of the liver and the kidneys. Therefore, the prevention and treatment of osteoarthritis in oriental medicine is aimed at more fundamental treatment based on these principles. Degenerative arthritis, one of the major bone joint diseases, is chronic arthritis. It is difficult to cure with anti-inflammatory drugs that are currently used in clinical trials as intractable diseases, and it also causes systemic side effects such as digestive disorders, gastrointestinal disorders, and renal function decrease. Long-term systemic administration in the treatment of many osteoarticular diseases has many problems. Therefore, the development of new drugs aimed at protecting and regenerative effects of anti-inflammatory and cartilage for the improvement and prevention of more aggressive symptoms than anti-inflammatory approaches such as anti-inflammatory action against degenerative arthritis that increases in proportion to the aging of the population. More urgent than ever Overseas, in addition to the development of anti-inflammatory drugs in recent years, with interest in articular cartilage protection, prevention of cartilage degradation and regeneration, joint histase inhibitors, free radical scavengers (active oxygen scavenging enzymes such as SOD), and joints Research on conservation therapy by long-term administration of tissue components (Chondroitin, Glucosamine etc.) has been actively conducted (Badger, AM, Cook, MN, et al., Journal of Pharmacology and Experimental Therapeutics, 290). , pp 587-593, 1999; Choi, JH, Choi, JH, et al., Osteoarthritis and Cartilage, 10 , pp471-478, 2002). Apart from this, recently, the development of arthritis treatments using herbal medicines has been actively conducted, and the development of medicines has been made. However, the situation has been mainly focused on suppressing the inflammatory response. Most of the arthritis treatments developed and used so far have been designed to focus on protecting and protecting cartilage and bone tissue of joint diseases, rather than inhibiting the inflammatory response, and in most cases, they have only limited therapeutic effects. there was.

On the other hand, in the Asian countries such as Korea, China, and Japan, the disease treatment agent has been used for thousands of years, and its efficacy has been proven, and the secretariat has been used in the private sector. Based on the selection of herbal medicines that can be developed based on the present invention, and the mechanism of the research through more rigorous basic research, a new concept of arthritis therapeutic herbal medicine was developed to complete the present invention.

The characteristics and specific actions of each of these herbs are as follows.

Single life (獨 活, Aralia cordata THUNB.) Is a perennial herb belonging to the Araliaceae, reaching 1.2 m in height with short hairs on the stem. The leaves are alternated with the leaves of 3 to 5 leaves, with the teeth on the edges of the leaves. The flower is light green, and the female flower and the male flower bloom in one branch, and in July-August, the inflorescence blooms at the end of the branch. Fruit is berry and ripens in October in black. It is often planted for use as a medicine. In the spring, the poisonous poison is used to treat migraines in the spring and autumn by cutting off the rhizome and roots and peeling the sun. As soon as the roots are dried in the sun, the fragrance will disappear, so it is better to dry them in a well-ventilated shade and dry them in the sun. The taste is spicy, warm and non-toxic, and has the effect of antipyretic, analgesic, myrrhosis, anti-inflammatory, blood coagulation promotion, cardiovascular action, coercive action. In the private sector, outposts are used as antipyretics, diuretics and analgesics. Root is used as a medicine. It is effective for dehumidification and tongjitong, so it is used to prescribe nasal pain, neuralgia, headache, and aftermath. Roots contain essential oils, while essential oils include limonen, sabnene, alpha-pinene, gamma-terpinene, myrcene, myrcene, Hugh mulren (humulene, α-caryophyllene), alpha-co pane (α-copaene), terpinene-4-ol is (terpinene-4-ol) is contained (jeongboseop and sinmingyo low, illustrated hyangyak (herbal) Dictionary, Younglimsa, p435, 1990).

Horseback Riding ( Cimicifuga heracleifolia Kom.) is a perennial herb belonging to the Ranunculuaceae, and its root stem is used for medicinal purposes. In oriental medicine, it has been known to have the effects of lifting, alveoli, fighting, and detoxification. The roots of horse riding include cimicifugine, salicylic acid, tannin, resin, caffeic acid, ferulic acid, cimitin, and alkaloids. ), Sugars, organic acids, glycosides, and isophorulic acid. (Gangso Medical Center, Chinese Medicine Dictionary , Book Publishing , Jungdam, pp3362 ~ 3372, 1998)

As described above, each of these herbal medicines is a medicine commonly used in various wounds or sudden chronic inflammation and anemia, headache, cold and detoxification, and each herbal medicine has its own pharmacological action and main effect.

However, until now, each of these herbal medicines has been studied only for its efficacy as a one-way herbal medicine and for the method of separating and preparing the active ingredient, and in some cases, a combination of several medicines have been attempted to see the therapeutic effect on arthritis. The efficacy was not scientifically proven and the effect was insignificant. Moreover, the focus was on efficacy in the cartilage protective effect underlying the treatment of arthritis.

The present inventors to maximize the properties and efficacy of each of these herbal medicines from the mixed herbs and to maximize the effect on inflammation-related diseases, in particular, the therapeutic effect and joint protection effect against arthritis, the traditional medicine based on the principle of oriental medicine Based on this study, 100 candidates were selected based on their activity to promote blood circulation and tissue regeneration, and based on these results, a screening test for the treatment of inflammation-related diseases was found. The present invention has been completed.

The present invention is to provide a pharmaceutical composition and health functional food useful for the prevention and treatment of inflammation-related diseases, in particular arthritis, containing a mixed herbal extract of venom and horse riding having an excellent cartilage tissue protection effect, anti-inflammatory and analgesic effect as an active ingredient. .

In order to achieve the above object, the present invention provides a pharmaceutical composition and health functional food for the prevention and treatment of inflammation-related diseases containing the extract of the poisonous and horse riding mixed herbal medicine as an active ingredient.

The mixed herbal extract is characterized in that the mixed dry weight ratio of poisonous and horse riding is 0.1 to 10: 1, preferably 0.5 to 5: 1, more preferably 1 to 2: 1, most preferably 1: 1 It is done.

The inflammation-related diseases include degenerative arthritis, rheumatoid arthritis, acute pain, chronic pain, joint pain, asthma, neuropathic pain, postoperative pain, migraine, neuropathy, nerve damage, irritable bowel syndrome, shock or inflammatory bowel caused by endotoxin Diseases and the like, preferably degenerative arthritis or rheumatoid arthritis.

The extract is water and a lower alcohol having 1 to 4 carbon atoms such as methanol, ethanol, butanol, or a mixed solvent thereof, preferably a mixed solvent of water and ethanol, more preferably 20 to 80% of a mixed solvent of water and ethanol Contains available extracts.

Hereinafter, the present invention will be described in detail.

Mixed herbal extract of the present invention can be obtained as follows.

About 1 to 20 of the weight of the sample prepared by mixing the poisoning and horse riding of the present invention in small portions and mixing them in an appropriate blending ratio, preferably in a dry weight blending ratio (w / w) of 0.1 to 10: 1, more preferably 0.5 to 5: 1. Times, preferably about 5 to 15 times the amount of water and lower alcohols of 1 to 4 carbon atoms, such as ethanol, methanol or the like, or a mixing ratio of about 1: 0.1 to 1:10, preferably 1: 0.2 to 1: 5 ( v / v) mixed solvent of water and ethanol, stirred extraction, hot water extraction, cold extraction, room temperature extraction, warm extraction, reflux cooling extraction or ultrasonic at room temperature for about 0.5 to 48 hours, preferably 1 to 15 hours. Extraction method such as extraction, preferably by stirring extraction, reflux extraction and warm extraction, and then the obtained extract is filtered, concentrated under reduced pressure or dried to obtain the poisonous and horse riding mixed herbal extract of the present invention.

In another aspect, the present invention provides a pharmaceutical composition for the prevention and treatment of inflammation-related diseases containing the poison and horse riding mixed herbal extracts obtained by the manufacturing process as an active ingredient.

Since the composition of the present invention has little toxicity and side effects, it can be used with confidence even for a long period of time for prophylactic purposes.

The composition for the prevention and treatment of inflammation-related diseases of the present invention comprises 0.1 to 50% by weight of the mixed herbal extracts based on the total weight of the composition.

The pharmaceutical composition comprising the mixed herbal extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The pharmaceutical composition comprising the mixed herbal extract of the present invention may be in the form of powder, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, according to a conventional method. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the mixed herbal extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, Calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compositions of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the composition of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably from 0.001 to 100 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

The composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

In addition, the present invention provides a health functional food for the prevention and improvement of inflammation-related diseases containing a poison and horse riding mixed herbal extract as an active ingredient.

As defined herein, "health functional food" means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to Act No. 6767 of the Health Functional Food Act, and "functional" means It means ingestion for the purpose of obtaining useful effects on health use such as nutrient control or physiological action on structure and function.

Examples of foods to which the mixed herbal extracts may be added include various foods, beverages, gums, teas, vitamin complexes, health functional foods, powders, granules, tablets, capsules or beverages.

It may also be added to foods or beverages for the purpose of preventing and treating inflammatory diseases. At this time, the amount of the extract in the food or beverage can be added in 0.01 to 15% by weight of the total food weight, the health beverage composition is added in a ratio of 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml Can be.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except the mixed herbal extracts as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. Can be. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (tauumatin, stevia extract (e.g., Rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the composition of the present invention may contain a natural fruit juice and a pulp for the production of fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Experimental Examples.

Comparative example  One. Poisonous  Extract manufacturer

It was purchased at Kyunghee Medical Center Oriental Medicine Hospital, and mixed with 1.5 L of 70% (v / v) ethanol to 200 g of shredded venom at a size of about 1.0 cm, and then refluxed for 4 hours. The filtrate was collected and collected, and 1.5 L of 70% (v / v) ethanol was added to the residue, followed by extraction for 4 hours, and the filtrates were combined and concentrated and lyophilized to 1 L to obtain 50 g of a powdered poison extract.

Comparative example  2. Horse Riding Extract Manufacturer

200 g of horse riding purchased from Kyunghee Medical Center Oriental Hospital was evenly mixed, and 1.5 L of 70% (v / v) ethanol was added thereto, followed by extracting under reflux for 4 hours while stirring well. The filtrate was collected and collected, and 1.5L of 70% (v / v) ethanol aqueous solution was added to the residue, followed by re-warming for 4 hours. The combined filtrates were concentrated to 1 L, and then freeze-dried to obtain 38 g of a powdered horse riding extract. .

Example  One. Poisonous  And Riding Mixed Herbal Extract Preparation

After mixing evenly 100 g of poison and 100 g of horse riding purchased from Kyunghee Medical Center Oriental Medicine Hospital, 1.5 L of 70% (v / v) ethanol was added thereto, followed by extracting under reflux for 4 hours while stirring well. The filtrate was collected and collected by adding 1.5 L of 70% (v / v) ethanol aqueous solution to the residue, followed by re-warming for 4 hours. The combined filtrates were concentrated to 1 L, and then freeze-dried to provide the powdery virulence and horse riding of the present invention. 44 g of the crude extract was obtained.

Experimental Example  1. Measurement of analgesic effect

1-1. Radiation-induced tail flick analgesia test

In order to examine the pain suppression effect of the mixed herbal extract of the present invention, a tail flick test was conducted using the method described in the literature as follows (Shaw FZ et al., Brain Res. , 911 (2)). , pp 105-115, 2001).

As a test animal, male ICR mice (Orient Bio, Japan) weighing 20 to 25 g were purified for several days, and then 8 animals were used in each group. After oral administration of the test drugs, infrared radiation was irradiated to the medial portion of the tail after 1 hour, and the time until the avoidance reaction was measured. The relative inhibition rate (%) was determined based on the inhibitory effect of the positive control group cocoxib. The calculation is shown in Table 1 below.

As shown in Table 1, the poison and horse riding mixed herbal extract of the present invention showed an excellent pain inhibitory effect than the single herbal treatment group.

1-2. Acetic acid-induced writhing

In order to investigate the analgesic effect on the mixed herbal extract of the present invention, the acetic acid-induced analgesic test was performed using the method described in the literature (BA Whittle, Brit. J. Pharmacol . , 22 , pp246-253, 1964). .

Male ICR mice (20-25 g body weight) were used as test animals in each group. After oral administration of the mixed herbal extract obtained in Example 1, pain was induced by intraperitoneal administration of 0.7% acetic acid (v / v in DW) 1 hour afterward. After recording the number of times, the inhibition rate (%) was calculated based on the number of twists of the treated group of celecoxib, the positive control group, as 100.

As shown in Table 2, the poison and horse riding mixed herbal extract of the present invention showed an excellent pain inhibitory effect than the single herbal treatment group.

1-3. Foot  Hot plate test

In order to investigate the analgesic effect on the mixed herbal extract of the present invention, a hot plate test was performed using the method described in the literature (Lanhers et al ., Planta) . Medica , 58 , pp 117-123, 1992).

The animals used male ICR mice (20-25 g) for 8 animals in each group. Experimental method was recorded 1 hour after oral administration of the test substance to measure the time until the mouse licks or jumps to the sole of the foot on the 55 ℃ hot plate, the inhibition rate (%) of the celecoxib treatment group When the average reaction time is based on 100, it is expressed as a relative value.

As shown in Table 3, the poison and horse riding mixed herbal extract of the present invention showed an excellent pain inhibitory effect than the single herbal treatment group.

1-4. Formalin analgesic experiment ( Formalin  analgesia test)

In order to investigate the analgesic effect on the mixed herbal extract of the present invention, a formalin animal model experiment was performed using the method described in the literature as follows (Fazli-Tabaei S et al., Behav . Pharmacol ., 16 , pp613-619). , 2005).

As the experimental animals, male ICR mice (Oriental Bio, Japan) weighing 20 to 25 g were purified for several days, and 8 mice were used in each group. Drugs for each treatment group were administered orally, and after 1 hour, 10% formalin solution (formalin, Sigma, USA) was subcutaneously administered to the left Fuji, immediately after injection for 5 minutes (1st phase), and 15 minutes to 20 minutes immediately after subcutaneous injection. Minutes (2nd phase) was recorded by measuring the time the mouse licking the soles, the inhibition rate (%) was expressed as relative values when the average time of the celecoxib treatment group was based on 100.

As shown in Table 4, the poison and horse riding mixed herbal extract of the present invention showed an excellent pain inhibitory effect than the single herbal treatment group.

1-5. Foot pressure  Paw pressure analgesia test

In order to investigate the analgesic effect on the mixed herbal extract of the present invention, a paw pressure analgesia test was conducted using the method described in the literature (Randall LO and Selitto JJ, Arch Int Pharmacodyn , 111, pp 409). -419, 1957).

As experimental animals, eight male SD rats (Oriental Bio, Japan) weighing 180-200 g were used after being purified for several days. Drugs for each treatment group were administered orally, 1 hour after 2% carrageenan (Sigma, USA) subcutaneously administered to the left Fuji, 3 hours after subcutaneous injection, using an analgesic meter (Ugobasile, Italy) The weight until the avoidance response due to pain was measured, and the inhibition rate (%) was calculated based on the weight of the celecoxib administration group and is shown in Table 5 below.

As shown in Table 5, the poison and horse riding mixed herbal extract of the present invention showed an excellent pain inhibitory effect than the single herbal treatment group.

Experimental Example  2. Anti-inflammatory effect measurement

2-1. Croton oil  cause Edema Anti-inflammatory  Experiment ( Croton  oil-induced ear oedema )

Croton oil, when applied to the skin, can cause redness, swelling, and blistering on the surface of the application. In order to evaluate the anti-inflammatory activity of the mixed herbal extracts of the present invention, experiments were performed using the method described in the literature using mice inducing edema with croton oil (Gabor M, Mouse ear indlammation models and their pharmacological applications , Published by Akademiai Kiado, Budapest, pp 24-28, 2000).

As experimental animals, six male mice (Orient Bio, Japan) weighing 20-25 g were used in each group. Treatment drugs were administered orally, and 1 hour later, 2.5% croton oil dissolved in acetone was evenly applied to the inside and outside of the right ear of the mouse to induce edema of the ear. After 4 hours of edema induction, mice were over-treated with ether, and then subjected to a rate change method (Patrick et al., Toxicol . Appl . Pharmacol . , 81, pp 476-490, 1985) using a thickness gauge . The degree of edema of the ears of the mouse was measured, and the inhibition rate (%) was calculated based on the weight of the celecoxib administration group and is shown in Table 6 below.

As shown in Table 6, the poison and horse riding mixed herbal extract of the present invention showed an excellent anti-inflammatory effect than the single herbal treatment group.

2-2. Arachidon acid-induced Edema Anti-inflammatory  Experiment ( Arachidonic  acid-induced ear oedema )

Arachidonic acid is a precursor of prostaglandins that causes platelet coagulation, arteriosclerosis, heart disease and inflammation, and is a major cause of inflammation. In order to evaluate the anti-inflammatory activity of the mixed herbal extract of the present invention, an experiment was performed using the method described in the literature using a mouse induced edema with arachidonic acid (Gabor M, Mouse ear indlammation). models and their pharmacological applications , Published by Akademiai Kiado, Budapest, pp 42-55, 2000).

As experimental animals, six male ICR mice (Orient Bio, Japan) weighing 20-25 g were used in each group. After 1 hour oral administration of treatment drugs, 50 mg / ml of arachidonic acid dissolved in acetone was evenly applied to the inside and outside of the right ear of the mouse to induce edema of the ear. After 1 hour of induction of edema, mice were over-treated with ether, and the degree of edema of the ears of the mice was measured by a speed change method using a thickness gauge, and the inhibition rate was determined based on the weight of the celecoxib administration group. %) Was calculated and shown in Table 7 below.

As shown in Table 7, the poison and horse riding mixed herbal extract of the present invention showed an excellent anti-inflammatory effect than the single herbal treatment group.

Experimental Example  3. Measuring cartilage protection effect

If the cartilage component, proteoglycan or collagen, cannot be produced quickly, and the cushioning function of the cartilage is lost, the cartilage is damaged and arthritis is caused. Articular cartilage consists of water (70-80%), collagen (10-15%), proteoglycans (5-10%) and chondrocytes for lubrication and growth.Proteoglycans are a single line of central protein ( It has a structure in which several glycosaminoglycans (GAGs) are attached to a core protein.

In order to confirm the joint protection effect of the mixed herbal extract of the present invention, the following experiment was performed.

3-1. Cartilage Tissue Culture

Articular cartilage was collected from the joints of 5 week old rabbits (Newzeland White Rabbit, Samtako Korea, Korea). After exposing the surface of the joint through sterile surgery, it took approximately 200-220 mg of the joint surface and inactivated the medium (heat treatment). 5% fetal bovine serum (FBS) and DMEM containing 100 units / ml of penicillin-streptomycin). The tissues were washed several times with medium and incubated for 1-2 days in a humidified 95% CO2 incubator at 37 ° C. Existing medium was replaced with basal medium (DMEM containing 100% / ml of 5% fetal bovine serum (FBS) inactivated by heat treatment, 10 mM HEPES and penicillin-streptomycin), and 30 mg of joint tissue was placed on a 48-well plate. After the transfer, the samples for each experimental group were processed. After incubation for 1 hour, 5 ng / mL of interleukin-1α was added to the medium to cause inflammation, and further cultured at 37 ° C. for 3 days. The supernatant was collected, transferred to fresh medium containing reagents, and incubated for another 25 days. Supernatants were collected at 3, 7, 14, and 28 days and stored at -20 ° C.

3-2. Determination of Glycosaminoglycan (GAG) Degradation

GAG constituting the proteoglycan using the 1,9-dimethylmethylene blue (DMB) assay to measure the protective effect of the mixed herbal extract of the present invention on the articular cartilage tissue of humans and rabbits In order to determine the degree of degradation, the following experiment was conducted using the method described in the literature (French MM et al., Ann. Biomed . Eng ., 32 (1) , pp 50-56, 2004).

The concentration of GAG in the culture medium of cartilage tissue obtained by the method of Experimental Example 3-1 was measured by the amount of polyanionic material produced by reaction with a Blyscan dye solution. Chondroitin sulfate was used. 50 μl of each test group was mixed with 500 μl of dye solution (Blyscan dye solution), reacted for 30 minutes at room temperature, and centrifuged at 12,000 rpm for 10 minutes. After dissolving the precipitate in the Blyscan dye-dissociation solution, the spectroscopic GAG amount was measured at 540 nm, and the percent inhibition was calculated based on the amount of GAG degradation induced by interleukin-1α (IL-1α). It is shown.

As shown in Table 8, the extracts of the poisonous and horse-riding mixed herbs of the present invention inhibited GAG degradation into the medium more significantly than the single herbal treatment group.

Experimental Example  4. Oral Toxicity Test

Acute toxicity experiments were carried out using a method described in the literature using four-week-old ICR mice (Orient Bio, Japan) (Haschek WM and Rousseaux CG, Handbook of toxicologic pathology , Published by Academic press, Inc., New York, pp 294-295, 1991).

In each group (5 / group), the mixed herbal extract of the present invention was dissolved and administered once orally at a dose of 5 g / kg / 10 ml. After administration of the mixed herbal extract of the present invention, the mortality, clinical symptoms, and weight change of the animals were observed, and hematological and blood biochemical tests were performed. It was.

As a result of the experiment, there were no clinical symptoms or dead animals in all animals to which the test substance was administered, and no toxicity change was observed in weight change, blood test, blood biochemistry test and autopsy findings. Therefore, the mixed herbal extract of the present invention does not show toxicological changes to 5,000 mg / kg in all mice, the minimum lethal dose (LD50) of oral administration was determined to be a safe substance more than 5,000 mg / kg.

Hereinafter, an example of the preparation of the composition containing the mixed herbal extract of the present invention will be described, but the present invention is not intended to limit the present invention, but is intended to be described in detail.

Formulation example  One. Powder  Produce

20 mg mixed herbal extract of Example 1

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation example  2. Preparation of Tablets

10 mg mixed herbal extract of Example 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation example  3. Manufacture of capsule

10 mg mixed herbal extract of Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation example  4. Preparation of Injectables

10 mg mixed herbal extract of Example 1

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na2HPO4,12H2O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation example  5. Liquid  Produce

20 mg mixed herbal extract of Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, lemon flavor is added to the mixture, and then the above ingredients are mixed, purified water is added to adjust the total amount to 100 ml, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation example  6. Manufacture of healthy food

1000 mg mixed herbal extract of Example 1

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

50 μg folic acid

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation example  7. Manufacture of health drinks

1000 mg mixed herbal extract of Example 1

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components in accordance with the conventional healthy beverage manufacturing method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated Used to prepare the healthy beverage composition of the invention. Although the composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

As described above, the composition containing the toxin and horse riding mixed herbal extract of the present invention as an active ingredient exhibits cartilage tissue protection effect, anti-inflammatory and analgesic effect, and thus is useful as a composition for preventing and treating inflammation-related diseases, especially arthritis. Can be used.

Claims (6)

Degenerative arthritis, Rheumatoid arthritis, Acute pain, Chronic pain, Arthralgia , containing mixed herbal ethanol extract with mixed dry weight ratio of Aralia cordata THUNB. And Horse riding ( Cimicifuga heracleifolia Kom.) As 0.1 to 10: 1 Pharmaceutical composition for the prevention and treatment of neuropathic pain, postoperative pain, migraine, irritable bowel syndrome or inflammatory bowel disease. delete delete delete Degenerative arthritis, Rheumatoid arthritis, Acute pain, Chronic pain, Arthralgia , containing mixed herbal ethanol extract with mixed dry weight ratio of Aralia cordata THUNB. And Horse riding ( Cimicifuga heracleifolia Kom.) As 0.1 to 10: 1 Dietary supplement for improving neuropathic pain, postoperative pain, migraine, irritable bowel syndrome or inflammatory bowel disease. delete