KR100429595B1 - Process for preparing composition comprising medicinal herb extract for preventing and curing arthritis and composition thereof - Google Patents

Abstract

The present invention relates to a method for producing a composition for treating and preventing arthritis, including a herbal extract, and a composition prepared by the method. The composition of the present invention, which is a pharmaceutical extract composition of Achyranthis root and Atractylodes japonica root, There is an effect, in particular, products such as fermentation thereof have a more excellent effect in the treatment of arthritis, can be used as an excellent effect of preventing and treating arthritis that can be used in place of the existing chemotherapeutic treatment.

Description

Process for preparing composition comprising medicinal herb extract for preventing and curing arthritis and composition

The present invention relates to a method for producing a composition comprising the extract of dew and producing extract showing an excellent effect on the prevention and treatment of arthritis and the composition. In particular, the present invention not only inhibits the production of tumor necrosis factor α (TNF-α) for the treatment of arthritis, but also transforms growth factor (TGF-β) and interleukin-4 (IL). -4) to promote the production of anti-inflammatory and collagen synthesis, such as the extract containing hyssop extract, creation extract, or mixtures thereof, which have an excellent effect on the regeneration of osteoblasts, cartilage tissue, etc. The present invention relates to a pharmaceutical composition for treating arthritis containing a product and a method of manufacturing the same.

In general, arthritis, or inflammatory disease of the joints, occurs in various forms, such as diseases associated with rheumatoid arthritis and joint inflammation.

Arthritis is a chronic polyarthritis that is characterized by inflammatory changes, especially in the synovial membrane of the lining of the articular capsule. It is a chronic progressive disease that causes swelling and pain in the joints of the whole body and, in severe cases, can become a physically handicapped person. In addition, arthritis diseases such as rheumatoid arthritis are progressive and develop joint disorders such as deformation and joint instability, often causing severe physical disorders due to lack of effective treatment and continued exacerbation.

The direct cause of arthritis is not clear yet, and research on this is ongoing. To date, there is no known effective prevention method for arthritis such as rheumatoid arthritis. There are many things to do.

Traditionally, these arthritis treatments include steroids such as cortisone and other corticosteroids, nonsteroidal anti-inflammatory agents such as aspirin, pyroxicam and indomethacin, gold agents such as orothioactive acid, chloroquinones and D-pheny Chemotherapy has been treated with various agents including antirheumatic agents such as silamine, gout inhibitors such as colchicine, and immunosuppressive agents such as cyclophosphamide, azathioprine, methotrexate and levamisol.

The treatments mentioned above are not fundamental therapies, and steroid hormones have been widely used as arthritis treatments, but their use has become difficult as their side effects become apparent.

In addition, arthritis, especially chronic rheumatoid arthritis, causes severe pain in patients, so be sure to take anti-inflammatory drugs, etc. Until now, aspirin and betazoline drugs have been widely used as drugs to alleviate these pains or remove joint swelling. Has been used. However, aspirin and the like have a fatal effect on the stomach, so it is difficult to continue taking the amount needed to treat arthritis.

Existing chemotherapeutic agents have drawbacks such as side effects that hinder the long-term use of the drugs, lack of anti-inflammatory effects, and lack of efficacy against arthritis that has already occurred. Shin, furpenic acid and non-steroidal anti-inflammatory agent is used to prepare a degree.

Therefore, there is a need for a solution to these problems and the development of arthritis therapies that have symptomatic effects on acute inflammatory symptoms and pain, and most of the arthritis drugs currently in use have some differences or personal differences. It is very important to develop medicines with fewer side effects, because they have side effects, and especially for the treatment of rheumatoid arthritis, it is necessary to take the drug for a long time.

In addition, some of the drugs may be administered by intravenous injection or intraperitoneal injection, in this case not only cumbersome, repeated intravenous injection, etc. may cause allergies, shock, as well as hygiene problems. Therefore, there is a need for a safe and easy to eat.

The present invention solves the above problems, and the object of the present invention is to provide a method for producing a composition for preventing and treating arthritis.

Another object of the present invention is to provide a composition prepared by the above production method.

1 is a diagram showing the volume change of the foot of the fermented drug (4B) treatment group and the control group over time after the injection of Mycobacterium tuberculosis.

Figure 2 is a diagram showing the volume change of the foot of the unfermented drug (1B) treated group and the control group with time after Mycobacterium tuberculosis injection.

Figure 3 shows the clinical Arthritic Index evaluation results of the drug treatment group and control group over time in arthritis-induced mice with type II collagen.

Figure 4 is a diagram showing the results of arthritis Incidence evaluation of the drug treatment group and the control group over time in arthritis induced mice with type II collagen.

Figure 5 is a graph showing the immunoglobulin (Immunoglobulin) concentration measured in plasma taken from the eye of the arthritis-induced arthritis treatment group and control rats with type II collagen.

Figure 6 is a comparison of the TGF-β concentration in each region of the isolated cells of the drug treatment group and control mice induced arthritis with type II collagen analysis by ELISA.

Figure 7 is a comparison of INF-γ concentrations in each region of the isolated cells of drug treatment group and control mice induced arthritis with type II collagen analysis by ELISA.

FIG. 8 shows electrophoresis of TGF-β after RT-PCR of cells collected from tissues of each group of arthritis-induced arthritis-treated and control rats with type II collagen.

9 is a diagram showing the electrophoresis of IL-4 after RT-PCR of cells collected from each site tissue of the arthritis-induced arthritis-treated group and control rats with type II collagen.

FIG. 10 is an electrophoresis picture of TNF-α after RT-PCR of cells collected from tissues of each group of arthritis-induced arthritis-treated and control rats with type II collagen.

FIG. 11 is a diagram comparing TNF-α concentrations between a control group and a hydrocortisone comparative drug group by treating each drug prepared in Example 18 in a macrophage of LPS-stimulated mice.

12 is a diagram comparing the inhibitory effect of Collagenase activity of the respective medicines prepared in Example 18.

In order to achieve the above object, there is provided a method for producing a composition for preventing and treating arthritis, comprising extracting the dew and creation and fermenting the extract.

The present invention preferably further comprises the step of concentrating after the fermentation step, and the solvent used for extraction of the present invention is preferably purified water, ethanol, methanol, propanol or butanol and a mixed solvent thereof, and purified water, ethanol or Methanol is more preferred.

In addition, it is preferable that the dew and the creation of the present invention are dried or herbal medicines. 0: 5-5: 0 is preferable, as for the ratio of the wedge and creation used for this invention, it is more preferable that it is 1: 10-10: 1, and it is especially preferable that it is 1: 2. At this time, in the case of using undried raw dew, the weight ratio is calculated in consideration of the included moisture content.

In the composition of the present invention, both the dew and the extract extract act as important medicines for the treatment of arthritis, but especially the inclusion of the dew and its fermentation products are more effective in inhibiting osteoclast activity and osteoblast activity such as TNF-α production inhibition and collagenase activity inhibition. Plays an important role in the treatment of arthritis.

In addition, the fermentation conditions of the present invention is less than 20 ℃ and less than 50 ℃ fermentation efficiency, it is preferred that 4 to 36 hours at 20-50 ℃, but gradually to 30-75 ℃ during the saccharification process to reduce the process time It warms up and includes the infusion method which puts a 52 degreeC protein resting period and a 65 degreeC glycosylation resting period for 30 minutes, respectively.

The present invention also provides a composition for preventing and treating arthritis prepared by the above method.

The composition of the present invention includes the extract of extracts, extracts and mixtures thereof, each of which may be extracted separately, it is a pharmaceutical composition containing a fermentation product produced by the extraction method by extracting together.

The composition of the present invention may further include conventionally used excipients, disintegrants, sweeteners, lubricants, flavoring agents, etc., tablets, capsules, powders, granules, suspensions, emulsifiers, syrups by conventional methods It may be formulated as a pharmaceutical preparation for unit administration or for several administrations, such as liquid or parenteral preparation. In the embodiment of the present invention it was formulated in the form of gel or soft extract.

The dosage level for a particular patient of the composition of the present invention may vary depending on the weight, age, sex, health condition, diet, time of administration, method of administration, excretion rate, severity of disease, and the like of the patient.

Arthritis in the present invention refers to all kinds of arthritis, and among them, especially means pulmonary arthritis, rheumatoid arthritis, degenerative arthritis, gouty arthritis, purulent arthritis and tuberculosis arthritis that are pulp by bacteria.

Other compositions of the present invention are also effective in autoimmune diseases such as systemic lupus erythematosus and amyloidosis.

Hereinafter, the present invention will be described.

Soemureup (Achyranthes fauriei LEVEILLE et VANIOT or Achyranthes bidentata Blume) is a perennial plant of about 90cm in height and amaranth (Amaranthaceae) plants. Roots ( wool , Achyranthis Bidentatae Radix ) contain saponins that break down into oleanolic acid and glucuronic acid when they are hydrolyzed, and akyrantins (C ₆ H ₁₁ O ₂ N · H ₂ O), which are loosened in water, and others Alkaloids, there is a large amount of mucus. The ash content of the water extract is about 8%, of which the potassium salt is 24.5%. In addition, there are amino acids including cytosterol, stigmasterol, aspartic acid, and polyvalent basic acids including succinic acid. Pharmacological effects include analgesic effect, antispasmodic effect, diuretic effect and anti-allergic effect. (Yook Chang-soo et al., Modern herbal medicine, Seoul, Hakchangsa, 24-128, 1993)

Cow knee root (wool) decoction has been used as a pill, urinating medicine, and pain medicine in motion therapy, and it is used as a commodity in neural herbaceous medicine and is used as a hematopoietic medicine in oriental medicine.

We used the present invention was purchased from the domestic ( Achyranthes japonica (MIQ.) NAKAI ) and Chinese erythromus ( Achyranthes bidentata Blume ), the roots of the postpartum uterus bleeding, various causes of uterine bleeding, menstruation It has been used to treat various kinds of gynecological diseases such as overdose.

Creation (Atractylodis Rhizoma) is refining the fine roots washed in water casserole in the spring or autumn as the rootstock of sapju (Atractylodes japonica KOIDZ.) Age years, ranging in height 80cm pull that dried in the sun, Asteraceae (Compositae) sapju in the perennial plant belonging to the It grows in mountains all over the place.

At the root (creation) of the insert, there are about 1.5% essential oils, carotene, inulin, rubbery, and alkaloid reactions, and outposts have saponin and coumarin reactions. The main components of essential oils are sesquiterpene atlactilone (C ₁₅ H ₂₀ O, melting point 38 ℃, about 20%), and furfural, β-Eudesmol, and henesol. Other sources include atlactilol, isovaleric acid esters, and atactic lactate potassium. In addition, the main ingredient of Atractylodes Koreana Kitam is the essential oil of atlanticodine and contains a small amount of acetylatactylodidinol, atlactilodidinol and elemol. There is very little or no atlactilone, the main ingredient for injectable essential oils.

Looking at the narrative data and pictures of old consent documents, the roots of Joseon shovel can be more appropriate to the creation.The creation is sweet and spicy and warm, so it has sedation, hematopoiesis, and urinary action. When urine is low due to disability, gastroenteritis, swelling, dizziness and pain when the whole body is urine, sweating medicine, etc. It is a very important medicine that is formulated in motion. In the private sector, it is said that long-term eating is used to stop diarrhea and vomiting, and often used for the treatment of dyspepsia, diabetes, cough and gout.

The present inventors confirmed that the extract of the dew and the mixture of the above-mentioned medicinal effects is a herbal composition having no poison and side effects and having an excellent therapeutic effect on arthritis, and the fermentation product of the composition thus prepared shows a marked increase in drug efficacy. It was confirmed to complete the present invention.

Accordingly, the product obtained by extracting water and alcohol respectively or together with water or alcohol, and the product obtained through the manufacturing process such as fermentation provides a pharmaceutical composition therapeutic agent of herbal composition with no side effects and excellent efficacy in preventing and treating arthritis. It may be usefully used as a substitute for chemotherapy.

The inventors of the present invention, although most arthritis treatments currently used to treat arthritis have a difference or a degree of individual difference as a chemotherapeutic agent, all have some side effects, so side effects that prevent long-term use of the drug, lack of anti-inflammatory effect In order to overcome the drawbacks such as the lack of efficacy against the already developed arthritis, herbal compositions for the treatment of arthritis of natural-derived herbal ingredients, which are considered to have relatively low side effects, have been studied for a long time. As a result, it was confirmed that the extract of the pharmaceutical herbal composition of Achyranthis Radix as the root of dew and Atractylodes japonica was effective in treating rheumatoid arthritis as a result of animal experiments, and the fermentation product of the extract showed better efficacy. Proved. At this time, by using the dry dew or natural raw dew to create and compounding conditions, extraction conditions and fermentation process to complete the present invention.

Therefore, the present invention relates to extract of pharmaceutical herbal composition of the hyssop and the production and its fermentation product, and also a method for producing the same. Preferably, rice, starch, etc. can be used in the fermentation process. The present invention relates to a pharmaceutical composition for preventing and treating arthritis such as concentrated extracts or pills, powders, capsules, and the like, and methods for preparing the same.

In addition, the present invention includes a concentrated extract from which the residual solvent is removed by extracting the dew with a lower alcohol having 1 to 4 carbon atoms, and the fermentation process may be applied to the extract thus prepared.

The present invention is investigated by administering to the rat the effect of preventing and treating arthritis of the pharmaceutical herbal composition extracts of hyssop and production. In addition, tissue cells from drug-treated and control rats were collected and analyzed for clinical evaluation and the concentrations of arthritis and inflammation-related cytokines, and in-vitro experiments were carried out on LPS-stimulated mouse macrophage (macrophage).

According to the present invention, as the root of the iron knee (Achyranthis ), the dew is Chinese Achyranthis bidentata BL . Using the dried dew or natural Achyranthis japonica (MIQ.) NAKAI domestic undried fresh dew , combined with the creation as the root of the Atractylodes japonica , a pharmaceutical herbal composition was extracted and then extracted with a solvent. Animal fermentation products obtained by fermenting malt or malt in fermentation were excellent in the prevention and treatment of arthritis.

The dew extract and the extract of the present invention can be extracted according to the following herbal extract method.

Place the hyssop and formation in the extraction solvent and warm at 50-120 ° C for 5-24 hours. The extract is cooled to 40-60 ° C. and filtered to take a supernatant, or the fermentation process used in the present invention can be applied without filtration. This extraction and filtration is repeated one or more times to combine the supernatant, and the solvent is concentrated by evaporation. The extracts can be extracted and used separately, but the extraction process is simple, and when warming, they can be heated in a 120 ℃ oil-bath or steam distilation, but preferably in a water bath at 80-100 ℃. Use the method of warming. In addition, when purified water was used as a solvent, the extraction amount was optimal at 12 times as a result of experiments under 6 to 16 times the dry extract usage.

Alternatively, the hyssop and the resulting product are added to an extraction solvent, cooled at room temperature or 4 ° C. for 5 to 7 days, and filtered to obtain a supernatant. This extraction and filtration is repeated one or more times to combine the supernatants, and the solvent is concentrated by evaporation.

In the two extraction methods as described above, it is preferable to grind the dew and the produce into small sizes. As an example of the extraction solvent, pure water or an aqueous alcohol solution such as 20 to 50% ethanol and 50 to 100% methanol can be used. The organic solvent is completely removed for application of the fermentation process of the invention.

Fermentation process used in the present invention can be carried out according to the following herbal fermentation method.

Put malt, yeast, etc. at 20 ~ 60 ℃ in the herbal extracts and ferment for 4 to 36 hours in the same temperature range. In this case, the herbal extract is obtained by the extraction method described above, and can be filtered after fermentation of the extract prepared by using the filtered supernatant or without the filtration process, if the herbal extract is filtered by fermentation without filtration process, the extract is filtered About 5% more fermentation product was obtained than when one supernatant was fermented and filtered.

Alternatively, the herbal extract may be slowly heated at 30 ° C., and a rapid fermentation method may be used by leaching, which has a protein resting period of 52 ° C. and a saccharification resting period of 65 ° C. for 30 minutes. In this case, it took 2 hours and 45 minutes to reach the final temperature of fermentation at 75 ° C, and the filtration time could be considerably shortened by high temperature filtration.

The present invention can proceed with fermentation by treating fermentation microorganisms such as natural fermentation or malt, yeast, grape and wine fermentation enzyme source or yeast. In particular, the fermentation of the composition of the present invention has a positive effect on the drug efficacy and taste, the shape of the final formulation (gel state) has the drug efficacy and storage advantages. In addition, the fermented rice with malt or malt, and then filtered to take a supernatant may be fermented into the herbal extract.

When using malt, a large amount of starch already contained by the malt enzyme can be saccharified to obtain an extract suitable for fermentation, and as a supplement, malt starch supplements are used up to 50% of malt. At this time, by using inexpensive side materials, economical fermentation efficiency, such as shortening fermentation time and increasing fermentation extraction amount, can be expected to have side effects such as flavor enhancement and turbidity reduction. Popular raw materials include refined starch of corn, rice, wheat, papain (from latex of Carica papaya), kiwi or pear (refined starch), and corns obtained by glycosylating these starches with acid or enzyme There are also glucose syrups, such as corn syrup.

The types of koji (koji) raw materials are wheat and rice coated with wheat flour, and mung bean, glutinous rice, and barley are also used. Yeast sources include air, dokmari, barley straw, straw, mulberry, wormwood, resident, lotus, young leaves of pine needles and pine needles. Non-raw stocks are roasted, spicy, steamed, and semolina. Raw materials have a grain size from 12% to fine powder. Types of auxiliary ingredients include mulberry leaves, mugwort, lactation decoction, peach seed powder, apricot seed powder, melon powder, and the like, which are prepared by treating coenzyme. Coenzyme enhances glycation by cultivating bacteria such as Rhyzopus, Usami, and 0ryzae, which are the main strains of conventional yeast, and simultaneously pursues the complex taste and saccharifying power of natural yeast.

The present invention will be described with reference to the following specific examples, but the present invention is not limited thereto. % As used herein and in the Examples means a percent by volume.

Example 1

85 g of fresh dew (50% water content) was washed well, and the dried and dried ginseng 85g was cut into small 2L round flasks, and 500 mL of water was placed in a reflux condenser, followed by steam extraction at 120 ° C. for 10 hours. The extract was cooled to about 75 ° C., filtered hot, the supernatant was collected, the solvent was evaporated to concentration, lyophilized to obtain 55 g of pills (preparation of 1B sample).

Example 2

After washing 85 g of fresh dew (50% water content) well and drying the dry and 85 g of dried radish, it was placed in a 2L round flask, and 500 mL of water was placed in a reflux condenser, followed by steam extraction at 120 ° C. for 10 hours. After the extract was cooled to about 50 ° C., 60 g of malt and 85 g of rice were added and fermented at 45 ° C. for 12 hours. This extract was filtered and the solvent was evaporated to concentration to obtain a flowable extract.

Lyophilization yielded 64 g of pills (preparation of 4B sample).

Example 3

Wash 40g of fresh dew (water content 50%) well and dry the dry and 40g of dried extract into a 5L round flask, and put 1280ml of purified water in a reflux condenser and extracted for 10 hours in a water bath at 80-100 ℃. After the extract was cooled to about 50 ° C., 12 g of malt and 32 g of rice were added and fermented at room temperature (warm temperature of about 30 ° C.) for 36 hours. At this time, when the rice floating in the liquid phase as the end point of the fermentation process, the extract was heated to 75 ℃ high temperature filtration and the solvent was evaporated to concentration to obtain a flowable extract of 85ml (sample of Experimental Example 2).

Example 4

20 g of dried dew (0.7% water content) and 40 g of dried extract were cut into small 2L round flasks, and 480 mL of water was placed in a reflux condenser and extracted for 10 hours in a 120 ° C oil bath. The extract was cooled to about 75 ° C., filtered hot, the supernatant was collected, the solvent was evaporated to concentration, and 31.5 g of solid was obtained. Compared with the result of Example 1, a crude extract of about 20% was obtained.

Examples 5-7

After washing 40g of fresh dew (50% water content) well and drying the dried and 40g of dried extract into 2L round flasks, purified water was put in a reflux condenser and extracted for 10 hours in a 120 ℃ oil-bath. The extract was cooled to about 50 ° C., and then 7g of malt was added thereto and fermented at 45 ° C. for 12 hours. This extract was filtered and the solvent was evaporated to concentration to obtain a solid.

Example 5 Example 6 Example 7 Purified water consumption 240 ml 320 ml 640ml Fermented Extraction 27 g 32.9 g 34.9 g

Example 8

20 g of dried dew (water content 0.7%) and 40 g of dried extract were cut into small 2L round flasks, 480 mL of water was placed in a reflux condenser, and extracted for 10 hours in a 120 ° C oil bath. After the extract was cooled to about 50 ° C., 7 g of malt and 3 g of starch were added thereto, followed by fermentation at 45 ° C. for 6 hours. This extract was filtered and the solvent was evaporated to concentration to give 36.8 g of solid content.

Example 9

20 g of dried dew (water content 0.7%) and 40 g of dried extract were cut into small 2L round flasks, 480 mL of water was placed in a reflux condenser, and extracted for 10 hours in a 120 ° C oil bath. The extract was cooled to about 75 ° C., filtered and the supernatant was collected, and 7 g of malt and 3 g of starch were added and fermented at 45 ° C. for 6 hours. This extract was filtered and the solvent was evaporated to concentration to give 34.6 g of solid.

Example 10

20 g of dried dew (water content 0.7%) and 40 g of dried extract were cut into small 2L round flasks, 480 mL of water was placed in a reflux condenser, and extracted for 10 hours in a 120 ° C oil bath. The extract was cooled to about 75 ° C. and filtered to collect supernatant. 7g of malt and 3g of starch were added, and 30-75 degreeC was rested for 30 minutes and 52 degreeC for 30 minutes, gradually raising temperature to 30-75 degreeC. After 2 hours and 30 minutes, the temperature of the solution reached 75 ° C. and left for 15 minutes, followed by filtration for 6 hours and fermentation for 6 hours in combination with the herbal extracts collected above. The solvent was evaporated to concentration to give 34.1 g of solid.

Examples 11-14

20 g of dried dew and 40 g of dried extract were cut into small 2L flasks, and 480 ml of 100% methanol solvent was extracted for 10 hours in an 80 ° C. water bath. This extract was filtered, the supernatant was taken, and the solvent was evaporated and concentrated to obtain a solid.

80 ml of purified water was added thereto, followed by fermentation for 6 hours.

Extraction solvent Extraction amount Fermented Extraction Difference Example 11 Methanol 480ml 11.63 g 3.1 g Example 12 ˝ 14.7g Example 13 480 ml of 30% Ethanol (Ethanol: water = 3: 7 v / v) 28.21 g 3.1 g Example 14 ˝ 31.33 g

Example 15

20 g of dried dew and 40 g of dried lanceolate were cut into small 2L round flasks, and 480 ml of 80% methanol (methanol: water = 4: 1 v / v) solvent was added thereto and cooled at 4 ° C. for 7 days. The extract was filtered, the supernatant was collected, the solvent was evaporated and concentrated to remove residual methanol completely, and 15.5 g of solid was obtained.

Example 16

100 g of malt (malt) and 100 g of fermented starch (starch), such as rice, wheat, corn, and papain, were added to 400 mL of purified water to prepare a fermentation broth by heating and leaching at 30 to 75 ° C. The solution is filtered hot at 75 ° C. to use a clear supernatant.

Example 17

The raw mill was selected, washed, sufficiently dried and ground to be mixed and kneaded while sprinkling about 20-25%. Put a certain amount into a yeast mold, press and mold, and finally apply the surface inoculation with a flour mixed with [greener greener mother-bearing Aspergillus oryzae ] and a white bacterium ( Asp. Nigar mut.Kawachii ). It was. It is placed on a steak covered with sterilized straw cushions in grains and covered with sterilized straw cushions. Afterwards, the product temperature was controlled to 35 ° C or lower, and then reversed every other day or every other day. After 4-5 days of granulation, yellowish green spores appeared and the temperature rose to 40 ° C or higher, and the surface of the grains began to dry. Afterwards, the temperature was gradually lowered and left for 8 to 10 days after incubation, aged for 7 days and stored in a dry storage room.

Example 18

Accurately weigh dry dew and produce and reflux extraction with 8 times (v / wt) methanol for 12 hours of use. The extract is filtered and the solvent is concentrated under reduced pressure. In the fermentation process, the fermentation broths prepared in Example 16 and Example 17 were mixed together and fermented at 45 ° C. for 12 hours.

Sample code Sample content Medicinal usage Lyophilization extract Fermented Extract an / h Fresh dew (water content 55%) 220 g 9.2 g ap / m Dry dew (moisture content 1%) 100 g 8.9 g b / m Creation 100 g 12.2 g an b / m Fresh dew / creation (220g / 100g) 20.6 ap b / m Dry dew / creation (100g / 100g) 21.8 an pro Fresh dew fermentation 220 g 23.1 ap pro Dry dew fermentation 100 g 18.1 b pro Creation fermentation 100 g 22.9 an b pro (Fresh dew / creation) fermentation (220g / 100g) 37 ap b pro (Dry dew / creation) fermentation (100g / 100g) 31.8 pro Fermentation broth 200 g 19.7 hydrocortison Reference - 25mg / 60Kg Usage

Example 19

Prepared in the same manner as in Example 8 to add an excipient and to obtain a granule, a powder was prepared as a hard capsule (Hard capsule).

Example 20

Prepared in the same manner as in Example 8 was mixed with a small amount of alcohol ethanol and purified water to prepare a soft capsule.

Experimental Example 1; Paw Volumn Change rating

The heat-treated Mycobacterium Butyricum was purchased from Difco, Detroit, MI, and male Wistar Lewis Rats were purchased from Charles River Japan Inc., and the mice were weighed at about 250 g after purchase (about 180 g). Other reagents, such as incomplete Freund adjuvant, were purchased from Sigma.

AA (acute arthritis) was used as a control by injecting Mycobacterium Butyricum (MB) into the frontal adjuvants (5 mg / ml) and injecting 100 μl per Wistar Lewis Rat into the subcutaneous layer of the right foot. In the same period, only the frontal adjuvants were subcutaneously injected. The volume of both feet was measured every 3-4 days by water displacement method and measured for about 28 days. The drug was administered at a dose of 2 g / kg based on solids and formed each group about 6-7 (Turull and Queralt, 2000; Immuno. Pharmacol. 46 (2000) 71-77.).

The names of the groups in the figure and the number of animals used are as follows.

Control Group: Control Group (6)

1B, 4B Group; Drug use experimental group (each seven)

Reference group (6)

Paw volume change was defined as the value of reducing the volume of the reference group in the control and experimental groups, respectively, and the AA inhibitory effect of the drug in the experimental group was defined by the following equation (Badger et al., Journal of Pharmacology and Experimental Therapeutics 291 (1999) 1380-1386).

In this experiment, 1B represents an unfermented medicinal herb extract ( Example 1 ), and 4B means a fermented medicinal herb extract ( Example 2 ).

Inhibitory Effect (%) = {1-[Experimental-Reference Group] / [Control-Reference Group]} × 100

For statistical analysis, the volume value of the experimental group was compared to the volume value of the control group using Student's t test.

the results are as follow.

Figure 1 summarizes the volume change of the foot over time after the MB injection. As shown in the figure, the first change was seen in the control group on day 3, which is 2.6 times higher than the reference date. The increase was maintained until day 16, but additional increase was observed at day 20 and increased to 3.7 times the initial volume. Afterwards, the trend changed to about 2.9 times as early as day 31. The general trend of 4B administration was similar to that of the control group, but the 4B group showed a different trend from the control group in that the volume of day 3 maintained or decreased until the end of the experiment. In terms of volume, 4B decreased to 55% of the control group (p> 0.001) at 20 days, showing a tendency to decrease than the control group at all time points. On the other hand, in the baseline group, the volume of the paw was maximum at the first three days (1,8 times greater than the first day) and then gradually decreased to recover to a slightly higher level than the initial value.

The mean weights of the experimental group, the control group and the reference group were almost the same as those of the 270g, but the weights decreased about 25g in the 4B, 1B and control groups at about 20 days after AA induction. On the other hand, the reference group showed an increase of about 60g. The left foot volume did not differ from the initial values in both groups.

As shown in FIG. 2, the administration of 1B showed a similar tendency to increase the volume initially and then increase again after a period of time (in this case, after 2 weeks). This tendency shows a clear difference from the tendency shown in the 4B administration of FIG. 1. In addition, 1B was found to have an effect in reducing edema compared to the control. On the other hand, 4B was found to have a superior effect to 1B in reducing edema compared to the control.

TABLE 1 Reduced right foot volume and significance compared to controls of 4B and 1B (p)

Date (4B) 0 3 6 9 13 16 20 23 25 28 31 Difference (ml) p value 0.08 (0.36) 0.20 (0.063) 0.65 (0.025) 0.46 (0.097) 0.68 (0.010) 0.69 (0.149) 1.31 (0.001) 1.14 (0.004) 0.71 (0.027) 0.51 (0.129) 0.62 (0.059) Date (1B) 0 3 5 8 14 17 19 23 25 28 Difference (ml) p value 0.01 (0.82) 0.38 (0.03) 0.6 (0.03) 0.6 (0.07) 0.55 (0.28) 0.55 (0.2) 0.73 (0.05) 0.89 (0.02) 0.77 (0.06) 0.72 (0.09)

(Bold letters indicate statistical significance with p value 0.05 or less)

Table 1 summarizes the differences from baseline values by measuring the edema of the right foot upon administration of 4B and 1B. As shown in FIG. 1, 4B showed a marked improvement in edema between 2 and 4 weeks compared to the control group, but the results of 1B administration showed a weak effect even though the effect was still on average.

[Table 2] Edema inhibition effect of 4B, 1B

Date (4B) 0 3 6 9 13 16 20 23 25 28 31 Inhibitory Effect (%) 20.4 30.1 22.9 26.6 28.2 44.1 40.6 28.4 23.8 29.0 Date (1B) 0 3 5 8 14 17 19 23 25 28 Inhibitory Effect (%) 58.5 34.1 24.5 18.2 5.75 22.0 27.7 28.0 29.0

(Bold letters indicate statistical significance with p value 0.05 or less)

Table 2 is a result of converting the above results into the inhibitory effect, and again shows that 4B drug administration is the best. Administration of 4B resulted in a reduction of about 40 to 45% of MB edema at 3 weeks, and the 1B drug showed 25% of this effect, so that the extract of fermented extract (4B) was not fermented. It showed higher therapeutic effect than extract (1B).

Experimental Example 2; Clinical Arthritic Index Assessment

Jackson Lab in the United States. 48 DBA / 1 experimental mice from the Korea Advanced Institute of Science and Technology (KIST) in 1988 were stabilized for 2 weeks with an average body weight of 21.5g, divided into two groups, the reference group and the experimental group, and then type II collagen. In order to cause arthritis (Type II Collagen-induced mouse arthritis test), the tail was immunized first. Two weeks later, the left hind paw was boosted in the same manner, and the experimental group 20 mice were then fermented with the drug extract of Example 3 fermented to the medicinal extract of Example 3 over 7 times for 2 weeks at a dose of 0.6 g / kg based on solids. (122.5 μl / 1 mice) orally. Observation of arthritis incidence and arthritis incidence over time in the experimental group (drug treatment group, Tolerance group) with respect to the comparative group (CIA group) was observed as the Clinical Arthritic Index (hereinafter referred to as "AI"). In Experimental Example 3, four rats were sacrificed two days after the booster, and a standard calibration curve was obtained to evaluate the therapeutic effect of the drug through histologic analysis. At the time point, the anti-inflammatory and arthritis treatment effects of the drug were analyzed by ELISA and RT-PCR methods through cell culture of the extracted tissues at the second sacrifice of four rats of the experimental group and the comparative group.

CFA (Complete Freund's adjuvant) reagent used for immunization was used with Arthrogon-CIA (5ml / vial Lot. 112700) from Chondrex, and Collagen Type II was mixed 1: 1 with CFA using Bovine native C II from Sigma. 100 μl per animal was injected into the tail of the rat. At the time of booster, IFA (Incomplete Freund's adjuvant) purchased from Difco was mixed 1: 1 with Bovine native C II, and 100 μl per animal was also injected into the rat's left hind foot fad. In the experiment, the degree of arthritis induction was the clinical arthritis index, and the degree of arthritis in all three feet except the left hind paw was observed and assigned 0 to 4 points according to the degree (0 point: no abnormal situation, 1 point: redness) Red swelling), 2 points: redness and edema expression, 3 points: spring due to the development of arthritis, swelling by edema as much as left hind paw, 4 points: severe inflammation, cracking, difficulty in stepping) % Represents the number of mice in which one of three paws can observe pronounced edema expression (above 3 points of AI evaluation).

Table 3 and Table 4 summarized the evaluation of AI and incidence by performing clinical evaluation from the 3rd week of feeding the medicinal herbs to the experimental group, which is shown in Figures 3 and 4. After 4 weeks of treatment, the control group (CIA group) showed the highest incidence of arthritis at 57.1%. In the drug treatment group, the incidence increased after the 6th week of drug administration. However, the incidence and degree of arthritis was hardly seen during the drug treatment period, thereby clearing and preventing the arthritis treatment. In addition, the AI evaluation analysis clearly shows the same tendency as the results of Experimental Example 1, in which the AI evaluation value increases first at 5 weeks and then increases at 2 weeks at 7 weeks. Since then, at the peak of AI evaluation, 66.4% of arthritis treatment effects were recorded for the CIA group, in which the drug treatment group did not exceed the average score of 1 point.

Table 3 Clinical Evaluation Chart CIA Induction Group

date Arthritic index Average Ⅰ group Group II Ⅲ group IV One 2 3 4 5 6 7 8 9 10 11 12 13 14 3rd week 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4-1 0 0 0 0 0 0 0 One One 0 0 One One 2 0.43 4-2 0 2 One 3 One 2 One 0 0 2 0 2 4 2 1.43 5 0 2 One 3 One One One 2 3 3 0 6 5 2 2.14 6 0 2 0 2 0 One One 6 3 2 One 4 5 3 2.14 7 3 2 One One 3 3 5 7 One 4 3 4 5 3 3.21 8 3 One 2 One 3 2 5 6 One 4 2 3 5 3 2.93 9 2 One One 2 2 2 4 5 One 3 2 2 4 3 2.43

date Incident Average Ⅰ group Group II Ⅲ group IV One 2 3 4 5 6 7 8 9 10 11 12 13 14 3rd week - - - - - - - - - - - - - - 0 4-1 - - - - - - - - - - - - - - 0 4-2 - - - - - - - - - - - + + - 14.3 5 - - - - - - - - + - - + + - 21.4 6 - - - - - - - + - + + + + - 35.7 7 - - - - - - + + - + + + + - 42.9 8 - - + - - - + + - + + + + + 57.1 9 - - - - - - + + - + + + + + 50

[Table 4] Clinical Evaluation Chart

date Arthritis index Average Ⅰ group Group II Ⅲ group IV One 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 3rd week 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0/16 = 0 4-1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 (2) 0/15 = 0 4-2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0/16 = 0 5 0 0 One 0 0 0 One One 0 0 One One One (One) 0 One 7/15 = 0.47 6 One 0 One 0 0 0 0 0 0 4 0 0 0 2 8/14 = 0.57 7 0 0 0 One One 2 4 0 2 0 One 2 13/12 = 1.08 8 0 0 0 One One 2 3 0 2 0 0 2 11/12 = 0.92 9 0 2 0 One 0 2 3 0 2 0 0 0 10/12 = 0.83

date Incident Average Ⅰ group Group II Ⅲ group IV One 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 3rd week - - - - - - - - - - - - - - - - 0/16 = 0 4-1 - - - - - - - - - - - - - - - (-) 0/15 = 0 4-2 - - - - - - - - - - - - - - - - 0/16 = 0 5 - - - - - - - - - - - - - (-) - - 0/15 = 0 6 - - - - - - - - - - + - - - + 2/15 = 13.3 7 - - - - - - - + + - - - - + 3/14 = 21.4 8 - - - - - + + - - - - + 3/12 = 25.0 9 - + - - - + + - - - - - 3/12 = 25.0

Experimental Example 3 Arthritis-induced Mouse Extracted Cell Histological Analysis

In animal experiment of Experimental Example 2 , after booster after 2 weeks, 4 mice of the reference group were sacrificed first before drug injection, and after 2 weeks, 4 mice of the control group (CIA group) and the drug treatment group after drug injection were completed. Tissue cells were extracted at the second sacrifice and cell cultures were measured to measure intracellular inflammation and immune-related cytokine concentrations by ELISA and RT-PCR, and anti-inflammatory and arthritis treatment effects were analyzed. In addition, the titration of immunoglobulin in the plasma collected from the eye was measured by antibody titration (antibody titration) to measure the total immune protein, IgG1, IgG2 and the degree of drug action on T cells.

Tissue cell extraction from rats was carried out at 4 sites of dLN (drainic lymph node), mLN (mensenteric lymph node), Spleen and PP (Peyer's Patches), and made cell soup in RPMI 1640 medium / 10% solution. Centrifuged (3000 rpm, 5 min) and seeded at 5 × 10 ⁵ cell concentration / well concentration. Enzyme Linked Immunosorbant Assay (ELISA) was used for transforming growth factor-β (TGF-β) and Interferon-γ (INF-γ) analysis using the SPECTA MAX 250 model, and RT-PCR (Reverse Transcription-Polymerase chain reaction) ), Perkin ELMER 9600 was used to measure the difference in TNF-α (Tumor necrosis factor-α), IL-4 (Interleukin-4) and TGF-β concentrations in tissue cells of CIA and Tolerance groups. The anti-inflammatory and arthritis treatment effects were compared and analyzed.

Table 5 and Table 6 show the ELISA analysis of TGF-β and INF-γ.In this case, the blocking solution was prepared using a PBS reagent of GIBCOBRL, 9.6 g of Dulbecco's Phosphate-Buffered Saline in 1L of purified water and 1% BSA. (TM-USB. Lot. 106268) It was made into the solution which added 10g and 0.1% sodium acetate. As antibodies for TGF-β measurement, # 244 (Catalog No. MAB1835) and # 19 (Lot No. WS04) of the US RD system were used, and # 17 (Cat. No. 100-21) of PEPRO TECH EC LTD (USA) was used. ) And # 18 (Cat. No. 80-1835) from Genzyme / Techne were used as standard and the catalog manual was referenced.

TABLE 5 ELISA PROCESS / TGF-β

1.Coating plates coated with # 244 with 4 ° C overnight (1/150 diluted-> (6μl + 894μl ccPBS) 3ml-> 50μl / well 2. Washing condition: PBST 1x 3. Blot the plate with blotting solution and incubate for 1 hour at room temperature 4. Washing condition: PBST 1x 5. Standard or Sample: Incubate Standard # 17 at room temperature for 2 hours (diluted: (2µg / ml)-> 2000pg, corresponding to 2µl + 238µl (blotting solution)-> 1/2 6. Washing condition: PBST 1x 7. Detect antibody # 19 and incubate for 1 hour and 30 minutes at room temperature (dilution: 1/250 1.8 μl + 898.2 μl (blotting solution) −> 50 μl / well 8. Washing condition: PBST 1x 9. Secondary enzyme: Streptavidin-HRP RD System. (Dilution: 1/200) Cultivation: 30 minutes on cow 10. Washing condition: PBST 1x 11. Substrate: Mixed with Substrate A: B = 1: 1-> 50µl / well (dilution) Culture: 30 minutes ~ 1 hour depending on the color development 12. Stop Solution: 1N HCl 13. Reading: 450nm Elisa Analyzer

[Table 6] ELISA Procedure / INF-r

1. Plates coated with # 352 were diluted to 4 ° C. overnight (1/500-> (3 μl + 2997 μl ccPBS-> 50 μl / well ccPBS). 2. Washing condition: PBST 1x 3. Blot the plates with blotting solution (diluted; 50 μl / well) and incubate for 1 hour at room temperature 4. Washing condition: PBST 1x 5. Standard or Sample: Standard # 344 incubated at room temperature for 2 hours (dilution: 1/100 2.4μl + 240μl (blotting solution)-> 1/2) 6. Washing condition: PBST 1x 7. Detect antibody # 353 and incubate for 1 hour and 30 minutes at room temperature (dilution: 1/250 3.6 µl + 900 µl (blotting solution) 8. Washing condition: PBST 1x 9. Secondary enzyme: Streptavidin-HRP RD System. (Dilution: 1/200-> 15 μl + 285 μl (blotting solution)) Incubation: 30 minutes in the dark 10. Cleaning conditions: PBST 1x 11. Substrate: Mixed with Substrate A: B = 1: 1-> 50µl / well (dilution) Culture: 30 minutes ~ 1 hour depending on the color development 12. Stop Solution: 1N HCl 13. Reading: 450nm Elisa Analyzer

ELISA analysis of INF-γ was also performed using RD system's # 352 (Cat. No. MAB785), # 353 (Cat. No. MAF485) and # 344 (Cat. No. 485-MI) as antibody standard reagents. See the manual.

Figure 5 is a graph showing the immunoglobulin concentration measured in plasma taken from the eye of arthritis-induced arthritis CIA group and drug-treated rats with type II collagen. 50 µl / Well coating of native bovine CII at 20 µg / ml concentration was performed, and the total immunoglobulin concentration and IgG1, IgG2 concentration in plasma were quantified using the detection reagents # 250, 182 and 183 purchased from Pharmingen. .

DBA / 1 experimental mice are immune proteins derived from T-help 2 cells (Th2) in the immunoglobulin in the body, which are involved in anti-inflammatory and antigen-induced inflammation by presenting antigens with Th1⇒IgG2 and total immunoglobulin. Lean concentration IgG did not differ, whereas IgG1 showed a more than double increase in drug treatment compared to control. That is, the drug treatment group specifically induced Th2 → IgG1 in T cells to induce anti-inflammatory action. (115.5% increase in IgG1 concentration)

ELISA analysis of TGF-β and INF-γ was performed on the extracted cells of the rats sacrificed secondarily and are shown in FIGS. 6 and 7. Two weeks after the booster, the mesenteric lymph nodes showed a significant increase in TGF-β (approximately 2.5-fold) in the torrent group compared to the CIA group, and the measurement of INF-γ, which is involved in inflammation, was also treated in mLN and dLN. The group obtained a decrease (20-40%).

TGF-β, which is present in osteoblasts, is inactive in bone matrix and is activated by acid released by osteoclasts during bone resorption to promote osteoblast proliferation and collagen synthesis and to inhibit osteoclasts. TGF-β is also known as monocaine (produced by macrophages), which is still commonly produced by bone or bone marrow cells and acts on the activation of epithelial and mesenchymal cells. ) As an inhibitory effect is an anti-inflammatory action.

In this experiment, the concentrations of TGF-β, TNF-α, and IL-4 were analyzed by RT-PCR and electrophoresed using 10 µg of cells of mLN, dLN, PP, and splenic tissues. Quantity unit: ng) RT-PCR method is a sensitive method for detecting and analyzing RNA expressed in cells, isolating specific RNA to be measured in the extracted cells, synthesizing c-DNA by reverse transcription and amplifying by PCR. Comparative analysis with control and calibration curve.

8, 9, and 10 are electrophoretic photographs of TGF-β, IL-4, and TNF-α analyzed by PCR, and measured in IOD values at 450 nm fluorescence detectors. It was.

Table 7 RT-PCR Results of TGF-β in Tissue Cells

mLN dLN Spleen P.P β2M result β2M result β2M result β2M result CIA County 20435 14852 0.73 14302 21081 1.47 6323 12230 1.93 8169 19263 2.36 Pharmaceutical treatment group 7622.1 19310 2.53 4002.5 19668 4.91 5340.5 17939 3.36 3509.4 13957 3.98 Difference 247% increase 234% increase 74% increase 69% increase

Table 8 RT-PCR Results of IL-4 in Tissue Cells

mLN dLN Spleen P.P β2M result β2M result β2M result β2M result CIA County 20435 9653 0.47 14302 4161 0.29 6323 2364 0.37 8169 8697 1.06 Pharmaceutical treatment group 7622.1 7404 0.97 4002.5 3469 0.87 5340.5 1237 0.23 3509.4 5447 1.55 Difference 106% increase 200% increase 38% reduction 46% increase

Table 9 RT-PCR Results of TNF-α in Tissue Cells

mLN dLN Spleen P.P β2M result β2M result β2M result β2M result CIA County 4953 15743 3.18 3537 9841.1 2.78 1737 10057 5.79 2496 7484.5 3.00 Medicinal treatment group 3379 6791.5 2.01 3329 8364.3 2.51 1908 10180 5.34 2680 5000.8 1.87 Difference 37% reduction 10% reduction 8% reduction 38% reduction

As a result, the amount of TGF-β recorded a 2.5-fold increase in booster position and mesenteric lymph nodes when the amount of change in IOD after PCR amplification was compared to the initial β2M IOD value of c-DNA, supporting the results of ELISA analysis. , Spleen and Peyer's Patch were also significantly increased, indicating that they are involved in anti-inflammatory and osteoblast proliferation (Table 7).

IL-4, which is involved in the activation of resting B cells as a B marrow cell stimulator, also shows an increase in the concentration of IL-4 by 100-200% in booster locations and mesenteric lymph nodes, indicating that they are involved in active B cell activation. The concentrations in the bone marrow-related organs, such as the spleen and PP, were not significantly different (Table 8).

Intracellular TNF-α is a tumor necrosis factor involved in inducing inflammation.

As a result, the concentration of TNF-α in spleen and Drainic LN did not show much difference, but it was significantly decreased in mesentery and P.P, indicating that it significantly influences the progression and differentiation of arthritis. In other words, the results of the post-boost incidence measurement in the drug treatment group, unlike the CIA induction group, it can be seen that significantly less AI and arthritis induction (Table 9).

Experimental Example 4 in-vitro experiment of mouse macrophages

The eleven samples of the medicinal herb prepared in Example 18 and the comparative drug were measured for the inhibitory effect of TNF-α and collagenase on LPS-stimulated cells in mouse macrophages with Hydrocortison. The results of the ELISA of TNF-α were significantly different between the LPS and negative groups, and there was no effect of the fermentation broth during the fermentation process of the medicinal herb. TNF-α inhibitory effect was recorded. In addition, the TNF-α inhibitory effect was also recorded in the sample group in which each extract was added to the fermentation process. It can be converted to support the results of Experimental Example 3 above.

It is shown in the figure in FIG.

The collagenase deactivation experiment was also measured by comparing the 10 samples of the hyaluronic acid produced in Example 18 above, the extract and the fermentation product of the extract and the fermentation broth itself to the negative control. First, each drug was added to a 0.5 ml solution containing 2% Azo dye-impregnated collagen (sigma), CaCl ₂ 1 nM, Tris-HCl 50 mM, Collagenase Type II (125 ng / 0.5 ml), and enzymes for 15 hours at 37 ° C. Reacted. UV / vis at 540 nm. The amount of azo dye separated by the reaction was measured by spectrophotometric comparison. As a result of this experiment, it can be seen that both the extract and the fermented extract of the medicinal herb showed an excellent effect on the inhibition of collagenase activity.

Table 10 and Figure 12 shows the picture.

[Table 10] Collagenase Inhibition Experiment-in-vitro Experiment

Medicinal name Enzyme + Herbs (average of two experiments) Medicine Activity with Inhibitors AN / B in MeOH 0.329 0.256 0.073 AP / MeOH 0.32 0.258 0.062 AP / B in MeOH 0.2745 0.243 0.0315 B MeOH 0.2425 0.228 0.0145 B pro in water 0.516 0.278 0.238 AP / B pro in water 0.489 0.259 0.2 AN pro in water 0.515 0.285 0.23 AN / B in water 0.492 0.268 0.22 AP pro in water 0.5365 0.283 0.2535 Pro 0.6075 0.304 0.3035 AN in water 0.3265 0.237 0.0895 Negative control 0.319

According to the present invention having the above-described configuration, the extract of hyssop and the extract also has an active part in alleviating the symptoms of arthritis, but in the case of the present invention which is a fermentation product of the extract, about 2 times as much as arthritis relief compared to the non-fermentation Showed effect.

In addition, in the present invention, the fermentation product of the above constitution is an arthritis preventive and therapeutic effect in the arthritis during the administration of the arthritis in the absence of any rate of arthritis prevention, the therapeutic effect also obtained a reduction of arthritis induction of 67%. In addition, by controlling the concentration of cytokines related to edema and arthritis in the body, it can be seen that it has an excellent effect not only for treating arthritis but also for treating edema.

Claims (18)

a) extracting the dew and creation; And b) a method of producing a composition for preventing and treating arthritis comprising fermenting the extract. The method of claim 1, Method for producing a composition for preventing and treating arthritis further comprising the step of concentrating after the fermentation step. The method of claim 1, wherein the solvent used for extraction is purified water, ethanol, methanol, propanol or butanol. The method of producing a composition for preventing and treating arthritis according to claim 1, wherein the ratio between the dew and the generation used in the extraction is 0: 5 to 5: 0. The method of claim 1, The dew drop and the creation is a method of producing a composition for preventing and treating arthritis, characterized in that the dry or herbal medicine. The method of claim 1, The urine may be Achyranthis japonica (MIQ.) NAKAI or Achyranthis bidentata BLUME , and the generation may be Atractylodes japonica KOIDZ. Or Atractylodes Koreana Kitam method for producing a composition for the prevention and treatment of arthritis. In step b) of claim 1, the fermentation uses one or more fermentation sources selected from malt, yeast, grape and wine fermentation enzyme sources and yeast, and as fermentation feedstocks such as rice or rice, corn starch, wheat, A method for producing a composition for preventing and treating arthritis, characterized in that papain, kiwi or pear is used directly, or starch thereof is purified or one or two or more of these glucose syrups are used in 0 to 50%. In the step b) of claim 1, the fermentation is arthritis, including fermentation source and fermentation subsidiary materials and coenzyme directly added to the medicinal herb extract or a fermentation broth, and also prepared by fermenting the combined medicinal herb extract respectively. Method of producing a composition for prevention and treatment. The method of claim 1, wherein the fermentation conditions are 2 to 36 hours at 20 ~ 75 ℃ method of producing a composition for the prevention and treatment of arthritis. A composition for preventing and treating arthritis prepared by the method of any one of claims 1 to 9. delete The method according to claim 1 or 2, Oral arthritis of extracts, pills, granules, powders and hard capsules or soft capsules thereof, comprising the fermented concentrate as a main active ingredient and using an appropriate amount of excipients in an effective amount and a pharmaceutically acceptable carrier Preventive and therapeutic pharmaceutical composition. The method of claim 10, The composition is a pharmaceutical composition, characterized in that to promote the production of the body TGF.beta. The method of claim 10, The composition is a pharmaceutical composition, characterized in that to inhibit the production of interferon-gamma in the body. The method of claim 10, The composition is a pharmaceutical composition, characterized in that to inhibit the production of Ti-F-alpha in the body. The method of claim 10, The composition is a pharmaceutical composition, characterized in that to promote the production of interleukin-4 in the body. The method of claim 10, The composition is a pharmaceutical composition, characterized in that to promote the production of immunoglobulin G1 mouse. The method of claim 10, The composition is for the purpose of treating and preventing inflammation.