KR20110006642A - Anti-arthritis and cartilage protective herbal preparations containing reduced cucurbitacin b - Google Patents

Abstract

PURPOSE: A herb composition containing herb extract including Trichosanthis radix for treating arthritis and protecting articulation is provided to suppress inflammation and to minimize cucurbitacin B content. CONSTITUTION: A herb composition for treating arthritis or protecting articulation contains Trichosanthis radix. The Trichosanthis radix extract contains 0.05 weight% of cucurbitacin B. A herbal composition contains herb extract of Clematis chinensis osbeck and Prunella vulgaris. A therapeutic agent for arthritis contains the herb composition as an active ingredient. The therapeutic agent is manufactured in the form of oral formulation, oral capsule, ointment, or injection.

Description

Anti-arthritis and cartilage protective herbal preparations containing reduced cucurbitacin B}

The present invention contains a herbal extract containing fruit muscles as an active ingredient, the extract is controlled to the content of cucurbitasin B causing side effects to less than 0.05% by weight relates to a safe arthritis composition for arthritis treatment and joint protection.

Fruit roots are well known as herbal medicines, and have been widely used for various inflammations such as boils, wounds, bronchitis, mastitis, tonsillitis, and gingiva. In addition, the gluteus maximus has the effect of eliminating moistness, cold and cold hands and feet, knee pain hurts, walking back and shoulders, the whole body is weak and has been used as a herbal medicine or herbal powder to treat symptoms such as pain . These symptoms are similar to those of general arthritis, including chronic rheumatoid arthritis in modern pathological concepts.

Fruit roots, also known as `` flower pots '' or `` white powders, '' are toxic herbal medicines that have been used to clean the roots of perennial vines, Hanultari and Yellow Hanultari, in the fall and to clean the outer shells, cut them into proper lengths, and use sun-dried herbs as herbs. to be. In oriental medicine, it stops thirst and is effective in various boils and dental fever mastitis. It has been widely used for drainage, small bowel, detoxification and antipyretic action. Trichosanthin, which is a protein, is the most widely known component of fruit muscles, and arginine, citrulline, etc. are known as amino acids, and palmitic acid and linoleic acid are fatty acids. (linoleic acid) and the like are known. Recently, stenols such as brinolic acid, 4-hydroxy-benzoic acid and alpha-spinasterol have been identified. , Korea Research Institute of Chemical Technology, pp 1354 ~ 1357 (1988); Doha Hyangje Ambassador, Younglimsa, pp960 ~ 963 (1990)].

Cucurbitacin B, on the other hand, is commonly found in Cucurbitaceae, such as cucumbers and melons. It has a bitter taste and can be commonly consumed around. However, high doses of cucurvitacin B may cause vomiting, diarrhea and gastrointestinal side effects, so care should be taken with extracts containing cucurvitacin B more than necessary. Fruit roots also contain cucurbitasin B corresponding to the gourd plants, accordingly it is necessary to adjust the content of cucurbitasin B in a range that does not cause adverse effects on the human body.

In particular, while maintaining the drug efficacy while reducing the content of substances causing side effects such as cucurbitasin B requires a lot of time and effort, which is a problem to be solved in the art for a long time.

On the other hand, the present inventors have received a patent for the fruit muscle composition for articular tissue protector, in which the active fraction obtained from the herbal extract of fruit juice is standardized and standardized with the content of 4-hydroxy-benzoic acid and vanillic acid. Patent 858,733.

However, there is no research on how to minimize the content of the cucurbitasin B or the minimum content that does not cause side effects.

Accordingly, the present inventors have conducted research and efforts to minimize the content of cucurbitasin B in extracts containing prior fruits, and as a result, when the suspension of extract powder containing fruits is heat-treated under certain conditions, the content of cucurbitasin B is greatly increased. The present invention has been completed by discovering that the present invention can be reduced, and thus minimized side effects.

The present invention is a herbal composition comprising an herbal extract containing fruit roots, characterized in that the herbal composition for treating or protecting arthritis, which contains less than 0.05% by weight of cucurbitasin B relative to the total extract.

The present invention also provides a herbal composition for treating arthritis or for protecting joints, wherein the herbal composition comprising the extract of the fruit extract of the fruit muscle, gastric gland, and hachichori contains Cucurbitasin B in an amount of 0.05% by weight or less based on the total extract. It is done.

Therefore, the present invention contains a herbal composition extracted from the fruit root muscle as an active ingredient, and the herbal composition for arthritis treatment and joint protection, and the arthritis treatment effect of the herbal composition, wherein the content of cucurvitacin B is adjusted to 0.05% by weight or less. The goal is to provide a way to reduce side effects without affecting them.

The extract of the present invention has excellent analgesic activity, inflammation inhibitory activity and articular tissue degrading enzyme inhibitory activity that the extract containing the conventional fruit muscle, while minimizing the content of cucurbitasin B causing side effects to various patients It is expected to be widely available.

1 shows the HPLC chromatogram of the fraction prepared by Example 4.

The present invention contains an herbal extract containing fruit muscles as an active ingredient, and the extract contains fruit juice composition for arthritis treatment and joint protection, wherein the content of cucurbitasin B causing side effects is adjusted to 0.05% by weight or less. It is done.

Hereinafter, the present invention will be described in detail.

Fruit roots contain 4-hydroxy-benzoic acid and vanillic acid, which are very effective as inhibitors of articular cartilage tissue degradation and joint tissue protection. . However, cucurbitacin B, which is present in the muscles of the muscles, can cause side effects of vomiting, diarrhea, and gastrointestinal tract when overdose. Therefore, the present invention controls the content of cucurbitasin B to 0.05 wt% or less, generally within the range of 0.001 to 0.05 wt%, to minimize the side effects while maintaining the intrinsic joint protective activity of the extract containing fruit muscles.

The preparation method of the herbal composition containing the fruit muscle according to the present invention includes the following process.

1) After reflux extraction with 7-15 times water or alcohol solution of herbal weight containing fruit roots, it is filtered, and the residue is added with 7-12 times water or alcohol solution of weight of herbal medicine, warmed and filtered And filtrate with

2) The filtrate obtained in 1) was separated into layers of 1 to 5 times lower alcohol or nonpolar solvent than the weight of the filtrate, and then concentrated under reduced pressure at 60 to 70 ° C.

3) azeotropically concentrated with 5 to 20 times the total amount of the extract obtained in 2) above, and homogeneously suspended in the same amount of water,

4) The suspension obtained in 3) above is heat-treated at 60-90 ° C. until the content of cucurbitacin B is 0.05% by weight or less, and lyophilized to prepare a powder extract.

The lower alcohol solvent used in the above production method may include isopropanol, propanol or butanol, and the like, and the nonpolar solvent may include ethyl acetate, dichloromethane, chloroform, carbon tetrachloride and methyl ethyl ketone, and the like using butanol. It is preferable from the viewpoint of activity and extraction efficiency.

Referring to the manufacturing process of the herbal composition containing fruit roots according to the present invention as described above in detail.

The present invention is standardized and standardized so as to have an active ingredient having excellent joint protection effect from a herbal composition mainly composed of fruit roots. The fruit roots used in the present invention are collected in the fall. In addition, the herbal composition may include gastrointestinal and hachicho in addition to fruit roots. The herbal composition is extracted by a hot water extraction method that has been conventionally used as a general bath, as well as extraction with water or an aqueous alcohol solution, and the filtrate thus obtained is subjected to layer separation with a lower alcohol and a nonpolar solvent.

Thus, water or alcohol aqueous solution is added to the herbal medicine containing fruit roots and refluxed for 2 to 5 hours. At this time, the amount of water or alcohol aqueous solution is appropriately 7 to 15 times the weight of the raw material of the herbal medicine. Then, the filtrate is collected by filtration, and the residue is added with an aqueous solution of water or alcohol of 7-12 times the weight of the crude ingredient, and then warmed again for 2 to 5 hours, filtered and mixed with the previous filtrate to increase the extraction efficiency. If the amount of water is too small, the stirring becomes difficult and the solubility of the extract is low, and the extraction efficiency is lowered. If the amount is too large, the amount of the lower alcohol and the nonpolar solvent used in the next purification step increases, which is not economical, causing problems in handling. Can be.

In the present invention, the method of re-extracting after the first extraction is adopted. Even in the case of mass production of the herbal extracts, even though the filtration is effective, the loss occurs because the water content of the herbal medicine itself is high, so the extraction efficiency is reduced only by the first extraction. This is to prevent this. In addition, as a result of verifying the extraction efficiency of each step, it was found that about 80 to 90% of the total extraction amount is extracted by the second extraction, and the third or more multistage extraction is considered to be economically low.

As described above, the extract obtained by extraction with an aqueous solution of water or alcohol over the first and second stages is filtered and concentrated, and then purified from impurities such as unnecessary proteins, polysaccharides and fatty acids contained in the filtrate. The impurities are purified by performing layer separation two to four times with 1 to 5 times lower alcohol or nonpolar solvent to obtain a solvent fraction. At this time, as the lower alcohol, butyl alcohol, propyl alcohol or isopropyl alcohol is used, and the nonpolar solvent is ethyl acetate, dichloromethane, chloroform, carbon tetrachloride and methyl ethyl ketone. When the amount of the lower alcohol or the nonpolar solvent is less than that of the filtrate, fine particles are formed by unnecessary components such as fatty acids, and thus the layer separation is not smooth and the extraction content of the active ingredient is not efficient.

The lower alcohol or nonpolar solvent fraction obtained after layer separation is concentrated under reduced pressure at 60 to 70 ° C. to remove the solvent remaining in the sample. The extract obtained after concentration is azeotropically concentrated two to three times with water of 5 to 20 times the total amount of the extract, and is then homogeneously suspended by adding the same amount of water again to obtain a suspension. The reason for azeotropic concentration with water at the time of the concentrated drying is to effectively control the content of the remaining lower alcohol in order to use the obtained herbal extract as a pharmaceutical raw material.

Thereafter, the suspension is heat-treated at 60-90 ° C. to reduce the content of cucurvitacin B to 0.05% by weight or less. If the heat treatment at a temperature of less than 60 ℃ significantly increases the time to lower the cucurbitacin B content, there is a problem that the extract is denatured when heat treatment above 90 ℃. The reason why the content of cucurvitacin B is decreased is that the natural extract shows acidity (pH 3 ~ 6) in aqueous solution, and it is believed that the hydrolysis of cucurvitacin B occurs actively when heat is applied.

The heat-treated suspension is freeze-dried to obtain an extract in powder form. The extract is excellent in inhibiting joint tissue degrading enzymes and protecting joints, while the content of cucurvitacin B is 0.05% by weight or less, and generally 0.001 ~. Since it exists in the range of 0.05% by weight to minimize the side effects, the herbal medicine using this extract is useful as a treatment for arthritis and joint protection.

Therefore, the present invention also includes a method of using a herbal composition containing the powder extract prepared as an active ingredient as an agent for treating arthritis and a joint protector.

It contains the powder extract obtained in the present invention, and is formulated by a conventional manufacturing method to prepare tablets, capsules, injections, and the like, of which lactose, microcrystalline cellulose, magnesium stearate, and the like, which are used as bases in the manufacture of tablets, are combined with the present invention. The use of extract in a ratio of 2: 1 can produce tablets that are active in treating arthritis and protecting joints.

In the preparation of such pharmaceuticals, herbal extracts may be used as such, but may also be mixed as pharmaceutically acceptable carriers, excipients, diluents, etc., into powders, granules, capsules, or injections. Manufacturing is possible. In addition, the herbal extract according to the present invention has been used for food and medicinal use since ancient times, and there is no particular restriction on its dosage, body absorption, weight, age, sex, health status of the patient, administration time, administration method, excretion Rate, the severity of the disease, and the like. In general, the herbal extract is preferably administered about 0.1 to 10 mg per 1 kg body weight. Therefore, the composition containing the active ingredient of the present invention is to be prepared in consideration of the effective amount range, and the unit dosage form formulated in this way according to the judgment of the expert and the needs of the individual to monitor or observe the administration of the drug as needed Specialized medications can be used or administered at regular intervals.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited to the examples.

Example  1: Preparation of herbal extract

The contaminants were removed, and 1,000 g of dried fruit juice was added to 7 L of 30 wt% aqueous ethanol solution and the mixture was refluxed for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 72 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form.

As a result of analyzing the herbal extract extracted and purified by the above method by HPLC under the following conditions, it was confirmed that the cucurbitacin B content was 0.01% (w / w).

1) Solvent

division 0 min 35 minutes 40 minutes Nitrile acetate-water (20: 80) 100% 0% 0% Nitrile acetate-water (45:55) 0% 100% 100%

2) Column: J'sphere (YMC) ODS-H80 (250 × 4.6 mm I.D., 4 μm)

3) Flow rate: 2.0 ml / min

4) Detector: UV 230nm

Example  2: Preparation of herbal extract

The contaminants were removed, and 1,000 g of dried fruit juice was added to 7 L of 30 wt% aqueous ethanol solution, followed by extraction under reflux for 6 hours with good stirring. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 60 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. As a result of analyzing the herbal extract extracted and purified by the above method by HPLC under the following conditions, it was confirmed that the cucurbitacin B content was 0.03% (w / w).

Example  3: Preparation of herbal extract

The contaminants were removed, and 1,000 g of dried fruit juice was added to 7 L of 30 wt% aqueous ethanol solution and the mixture was refluxed for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 48 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. The herbal extract extracted and purified by the above method was analyzed by HPLC under the following conditions.

Example  4: Preparation of herbal extract

The contaminants were removed, and well-dried 250 g of the gastric glands, 500 g of fruit roots, and 250 g of Hagocho were mixed well, and 7 L of 30 wt% aqueous ethanol was added thereto, followed by extraction under reflux for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 60 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. As a result of analyzing the herbal extract extracted and purified by the above method by HPLC under the following conditions, it was confirmed that the cucurbitacin B content was 0.01% (w / w), and the chromatogram analyzed by HPLC is shown in FIG. 1. Shown in

Example  5: Preparation of herbal extract

The contaminants were removed, and well-dried 250 g of the gastric glands, 500 g of fruit roots, and 250 g of Hagocho were mixed well, and 7 L of 30 wt% ethanol solution was added thereto, followed by extraction under reflux for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 48 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. The herbal extract extracted and purified by the above method was analyzed by HPLC under the following conditions.

Example  6: Preparation of herbal extract

The contaminants were removed, and well-dried 250 g of the gastric glands, 500 g of fruit roots, and 250 g of Hagocho were mixed well, and 7 L of 30 wt% ethanol solution was added thereto, followed by extraction under reflux for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 36 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. The herbal extract extracted and purified by the above method was analyzed by HPLC under the following conditions.

Comparative example  1: Preparation of herbal extract

The contaminants were removed, and 1,000 g of dried fruit juice was added to 7 L of 30 wt% aqueous ethanol solution and the mixture was refluxed for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 36 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. As a result of analyzing the herbal extract extracted and purified by the above method by HPLC under the following conditions, it was confirmed that the cucurbitacin B content was 0.07% (w / w).

Comparative example  2: Preparation of herbal extract

The contaminants were removed, and 1,000 g of dried fruit juice was added to 7 L of 30 wt% aqueous ethanol solution and the mixture was refluxed for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L of distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 50 ° C. for 24 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. As a result of analyzing the herbal extract extracted and purified by the above method by HPLC under the following conditions, it was confirmed that the cucurbitacin B content was 0.09% (w / w).

Comparative example  3: Preparation of herbal extract

The contaminants were removed, and 500 g of well dried fruit root was added to 7 L of 30 wt% aqueous ethanol solution, followed by extraction under reflux for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrated concentrate, and the suspension was lyophilized to obtain a crude herbal extract in powder form. As a result of analyzing the herbal extract extracted and purified by the above method by HPLC under the following conditions, it was confirmed that the cucurbitacin B content was 0.13% (w / w).

Comparative example  4: Preparation of herbal extract

The contaminants were removed, and well-dried 250 g of the gastric glands, 500 g of fruit roots, and 250 g of Hagocho were mixed well, and 7 L of 30 wt% ethanol solution was added thereto, followed by extraction under reflux for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 24 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. The herbal extract extracted and purified by the above method was analyzed by HPLC under the following conditions.

Comparative example  5: Preparation of herbal extract

The contaminants were removed, and well-dried 250 g of the gastric glands, 500 g of fruit roots, and 250 g of Hagocho were mixed well, and 7 L of 30 wt% ethanol solution was added thereto, followed by extraction under reflux for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L distilled water was added to the concentrated azeotropic concentrate, which was suspended. The suspension was subjected to a heat treatment at 80 ° C. for 12 hours, and the suspension was lyophilized to obtain a crude herbal extract in powder form. The herbal extract extracted and purified by the above method was analyzed by HPLC under the following conditions.

Comparative example  6: Preparation of herbal extract

The contaminants were removed, and well-dried 250 g of the gastric glands, 500 g of fruit roots, and 250 g of Hagocho were mixed well, and 7 L of 30 wt% ethanol solution was added thereto, followed by extraction under reflux for 6 hours while stirring well. The filtrate was collected and collected. The residue was re-extracted for 3 hours by adding an aqueous solution of 30 wt% ethanol, and the filtrates were combined and concentrated to 1 l. Two times the saturated n-butyl alcohol twice the weight of the concentrated filtrate was added thereto, followed by two layer separations, and only the n-butyl alcohol layer was collected and concentrated under reduced pressure until the herbal extract was dried at 60 to 70 ° C. Most of n-butyl alcohol and water were evaporated and 1 L of distilled water was added to perform azeotropic concentration, which was repeated twice more. 1 L of distilled water was added to the concentrated azeotropic concentrated concentrate, and the suspension was lyophilized to obtain a crude herbal extract in powder form. The herbal extract extracted and purified by the above method was analyzed by HPLC under the following conditions.

Experimental Example  1: Analgesic effect test

In order to compare the activity of the extract prepared by Examples 1 to 6 and Comparative Examples 1 to 6 for the analgesic action, an acetic acid-induced guiding model test was carried out by the following experimental method and the results are shown in Table 2 below. As shown.

Experimental Method

ICR (Institute of Cancer Research) rats extracts prepared by the methods of Examples 1 to 6 and Comparative Examples 1 to 6 orally administered 200 mg or 400 mg per kg of mice 0.6 hours (v / v) after 1 hour ) A peritoneal injection of acetic acid at a dose of 0.1 ml per 10 g of rat's weight, and the pain response of each rat for 10 minutes from 10 minutes after injection. Writhing: Stretching back or stretching the hind legs completely behind the body ) Was observed.

Extract Dose (mg / kg) Average number of rides Suppression rate
(%) Control - 19 - Example 1 200 12.0 36.8 400 9.0 52.6 Example 2 200 11.0 42.1 400 8.0 57.9 Example 3 200 10.6 44.2 400 7.9 58.4 Example 4 200 9.1 52.1 400 6.9 63.7 Example 5 200 9.5 50.0 400 7.1 62.6 Example 6 200 9.6 49.4 400 7.0 63.2 Comparative Example 1 200 10.1 46.8 400 7.0 63.2 Comparative Example 2 200 10.0 47.4 400 7.1 62.6 Comparative Example 3 200 9.8 48.4 400 7.0 63.2 Comparative Example 4 200 9.3 51.1 400 6.8 64.2 Comparative Example 5 200 9.8 48.4 400 7.4 61.1 Comparative Example 6 200 10.0 47.4 400 7.0 63.2

As shown in Table 2, the extracts of Examples 1 to 6 and Comparative Examples 1 to 6 were all analgesic activity, it was confirmed that there is no significant activity difference between each other.

Experimental Example  2: test the inhibitory effect on acute inflammation

The inhibition of Croton oil-induced acute inflammation of extracts prepared in Examples 1 to 6 and Comparative Examples 1 to 6 was compared, and the degree of inhibition of ear edema induced by croton oil was compared with that of the control group. It is expressed as a percentage by the results are shown in Table 3 below.

Experimental Method

ICR (Institute of Cancer Research) rats were orally administered the herbal extracts obtained in Examples 1 to 6 and Comparative Examples 1 to 6, and after 1 hour, 0.025 ml of 2.5% croton oil was evenly applied to the right ear, and 3 Left and right ear edema was measured after the passage of time.

Extract Dose (mg / kg) Average inflammation rate (%) Inhibition Rate (%) Control - 85.00 - Example 1 200 70.52 17.0 Example 2 200 72.93 14.2 Example 3 200 70.89 16.6 Example 4 200 68.72 19.2 Example 5 200 67.33 20.8 Example 6 200 66.95 21.2 Comparative Example 1 200 69.53 18.2 Comparative Example 2 200 69.51 18.2 Comparative Example 3 200 69.17 18.6 Comparative Example 4 200 66.94 21.2 Comparative Example 5 200 67.17 21.0 Comparative Example 6 200 67.85 20.2

From the results of Table 3, the extracts of Examples 1 to 6 and Comparative Examples 1 to 6 were found to have excellent inflammatory inhibitory activity as the edema rate was reduced, but it was confirmed that there is no significant activity difference between each other.

Experimental Example  3: Test of the Effect of Inhibiting Articular Tissue Enzyme Activity

Examples 1 to 6 and Comparative Examples 1 to 6 was performed a comparative experiment on the inhibitory action of hyaluronidase (Hyalulonidase), which is an articular tissue degrading enzyme, and the results are shown in Table 4 below.

Experimental Method

After activating the hyaluronidase by incubating at 37 ° C. in acetate buffer for 20 minutes, potassium hyaluronate was added as an extract and substrate obtained in Examples 1 to 6 and Comparative Examples 1 to 6 for about 40 minutes. Continue to incubate. The reaction was terminated with NaOH, potassium borate (potassium borate) was added, heated to 100 ° C. and developed with DMBA (Dimethylbenzanthracene) to measure the absorbance and the inhibition rate was calculated in comparison with the control.

Test concentration (mg / mL) Inhibition Rate (%) Example 1 1.0 82.3 Example 2 1.0 83.7 Example 3 1.0 89.6 Example 4 1.0 92.5 Example 5 1.0 91.6 Example 6 1.0 91.8 Comparative Example 1 1.0 80.5 Comparative Example 2 1.0 79.1 Comparative Example 3 1.0 82.5 Comparative Example 4 1.0 89.8 Comparative Example 5 1.0 91.7 Comparative Example 6 1.0 90.6

As shown in Table 4, it was found that the extracts of Examples 1 to 6 and Comparative Examples 1 to 6 were all excellent in inhibiting the activity of articular tissue degrading enzymes, but there was no significant difference in activity between them. .

Experimental Example  4: mouse Single dose  Toxicity test

Experiments whether the extracts prepared by Examples 1 to 6 and Comparative Examples 1 to 6 exhibit acute toxicity are shown in Table 5 below.

Experimental Method

ICR (Institute of Cancer Research) rats were orally administered 2 g / kg of the extracts of Examples 1 to 6 and Comparative Examples 1 to 6 and observed whether or not death occurred. (10 mice per group)

Test concentration (g / kg) Lethality (%) Autopsy Results Example 1 2.0 0 No specific symptoms Example 2 2.0 0 No specific symptoms Example 3 2.0 0 No specific symptoms Example 4 2.0 0 No specific symptoms Example 5 2.0 0 No specific symptoms Example 6 2.0 0 No specific symptoms Comparative Example 1 2.0 10 + Comparative Example 2 2.0 20 +++ Comparative Example 3 2.0 30 +++++ Comparative Example 4 2.0 10 ++ Comparative Example 5 2.0 20 ++++ Comparative Example 6 2.0 30 +++++

※ +: The extent of lesions in the digestive system, the more +, the more severe the lesion.

As shown in Table 5, the administration of the extract of the example of the content of the cucurbitasin B is 0.05% (w / w) or less, but no specific symptoms appeared, but the cucurbitasin B content is more than 0.07% (w / w) When the extract of the comparative example was administered, lesions appeared in the gastrointestinal tract, and it was confirmed that mortality appeared. In particular, it was confirmed that the mortality increased and the degree of lesions increased as the content of cucurvitacin B increased.

Therefore, the extract of the present invention has excellent analgesic activity, inflammation inhibitory activity and articular tissue degrading enzyme inhibitory activity, but did not show side effects, it was confirmed that it is useful as an arthritis treatment and joint protection agent.

Production Example  1: preparation of tablets

Tablets for oral administration were prepared using the wet granules method and the dry granules method using the herbal extract of Example 1 in the following composition.

[Furtherance]

Herbal extract 200 mg, light silicic acid anhydrous 10 mg, magnesium stearate 2 mg, microcrystalline cellulose 50 mg, starch glycolate 25 mg, corn starch 113 mg, ethanol anhydride.

Production Example  2: Preparation of Ointment

Using the herbal extract of Example 1 was prepared an ointment with the following composition.

[Furtherance]

Herbal extract 5 g, cetyl palmitate 20 g, cetanol 40 g, stearyl alcohol 40 g, myristan isopropyl 80 g, monostearic acid sorbitan 20 g, polysorbate 60 g, paraoxybenzoic acid propyl 1 g, para 1 g of methyl oxyanate, phosphoric acid and purified water

Production Example  3: Preparation of Injection

Using the herbal extract of Example 1 was prepared an injection with the following composition.

[Furtherance]

Herbal extract 100 mg, mannitol 180 mg, sodium dihydrogen phosphate 25 mg, water for injection 2974 mg

Production Example  4 : Transdermal  Produce

Using the herbal extract of Example 1 to prepare a transdermal drug in the following manner.

[Composition 1]

Herb extract 0.4 g, sodium polyacrylate 1.3 g, glycerin 3.6 g, aluminum hydroxide 0.04 g, methyl paraben 0.2 g, water 14 g.

[Composition 2]

0.8 g of herbal extracts, 1.6 g of propylene glycol, 0.8 g of liquid paraffin, 0.4 g of isopropyl myristate, 16.4 g of gel bar 1430.

Claims (8)

Herbal medicine composition comprising an herbal extract containing fruiting root, Cucurbitasin B is contained in 0.05% by weight or less based on the total extract for the treatment of arthritis or arthritis herbal composition.
Herbal medicine composition comprising an herbal extract of fruit roots, gastrointestinal tract, and hawthorn, wherein the cucurbitasin B is contained in an amount of 0.05% by weight or less based on the total extract.
The herbal composition of claim 1 or 2 is contained as an active ingredient for the treatment of arthritis, and a therapeutic agent for arthritis, characterized in that it contains 0.001 to 0.05% by weight of cucurbitasin B based on the total extract.
4. The therapeutic agent for arthritis according to claim 3, which is formulated as an oral tablet, oral capsule, ointment, injection or transdermal administration.
The herbal composition according to claim 1 or 2, which is contained as an active ingredient for joint protection, and a joint protecting agent, which contains 0.001 to 0.05% by weight of cucurbitasin B based on the total extract.
The joint protector according to claim 5, which is formulated as an oral tablet, oral capsule, ointment, injection or transdermal administration.
A method for reducing the side effects of fruit extracts without affecting the therapeutic effect of arthritis of fruit extracts, characterized in that the content of the cucurbitasin B is adjusted to 0.05% or less of the total weight of the extract.
Fruit extracts of fruit juice, gastric juice and haejucho extract, characterized in that the content of the cucurbitasin B is adjusted to 0.05% or less of the total weight of the extract, without affecting the arthritis treatment effect To reduce side effects.

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