KR20120086262A - Fraxinus rhynchophylla Hance extracts compositions for treating or preventing inflammatory diseases - Google Patents

Abstract

PURPOSE: A composition containing Fraxinus rhynchophylla Hance extract is provided to effectively suppress inflammation and acute and chronic inflammation. CONSTITUTION: A pharmaceutical composition for preventing and treating inflammatory diseases contains Fraxinus rhynchophylla Hance extract as an active ingredient. The extract contains 1.0-0.5 wt% of caffeine as an indicator. The inflammatory diseases include arthritis, adema, and immune diseases. A method for preparing the extract comprises: a step of extracting Fraxinus rhynchophylla Hance with water, alcohol or an alcohol solution, cooling and filtering; a step of concentrating the liquid at 50-90 °C; and a step of adding alcohol and centrifuging at 700-1,200 rpm.

Description

{Fraxinus rhynchophylla Hance extracts compositions for treating or preventing inflammatory diseases}

The present invention relates to a composition for treating and preventing inflammatory diseases comprising a dermis (秦 皮, Fraxinus rhynchophylla Hance) extract as an active ingredient, more specifically, the standardization and so that the content of caffeine in the dermis extract to a certain range The dermis extract can be used as an agent useful for the treatment and prevention of diseases caused by inflammatory changes such as arthritis due to the excellent expression of inhibitory effects caused by inflammatory changes such as analgesic suppression, acute inflammation suppression, and acute edema suppression. It is about.

Arthritis is a medical term that collectively refers to the development of inflammatory changes in the joints by some cause, such as bacteria or trauma. Such arthritis is divided into acute and chronic.

Acute arthritis is classified as follows: ① serous (염 性) arthritis: Usually caused by trauma, the cause is unknown, usually occurs in only one joint. ② serous fibrosis (奬 液 纖維素 性) arthritis: occurs during acute arthritis of arthritis, turbid effusion in the joint cavity (渗出 液) stops. Fibrosis of the stomach membrane (생) is created, even if the inflammation subsides, leaving severe movement disorders. ③ purulent (化膿 性) arthritis: joints of open window (開放 創) or gonorrhea, typhoid fever, scarlet fever, sepsis (보인다) is a multiple of infectious diseases. Infants 1 to 2 months of age cause severe bone damage, resulting in incurable dislocation. In adults, periosteal osteomyelitis causes the peas to burst and pus enters the joints. This is called secondary pyogenic arthritis.

Chronic arthritis is classified as follows: ① Specific Inflammation: Tuberculosis, syphilis or postmenopausal arthritis due to many metabolic disorders of uric acid in middle-aged men. ② Multiple arthritis: Chronic arthritis is caused by many cases of acute serous arthritis (移行) or tuberculosis, syphilis, gonorrhea during the course of the multiple, one of the sepsis. In addition, it also includes arthritis called Still's disease. ③ deformed osteoarthritis: bone or joint aging or trauma is the cause. ④ hemophilia (血友病 性) arthritis: when hemophilia is caused by bleeding in the joints.

Many drugs have been developed for the purpose of treating diseases caused by inflammatory changes typified by arthritis as described above.

On the other hand, the dermis (秦 皮, Fraxinus rhynchophylla Hance) is the name of the Chinese herbal bark (樹皮), the deciduous broad-leaved arborescent of the ash family. Its height is 10m and the eggplant is grayish brown and hairless. The leaves are the rider's right upper lobe and face each other. The leaflets are 6 ~ 15cm long, broad egg, wide basal, 5 ~ 7, the front of the leaf is green, hairless, grayish green, hairy, At the edge there is a wave-shaped mount or flat. The flowers are inverted egg shape in May and run in conical or abdominal inflorescence, and the calyxes which split into 4 are almost flat. Fruits are poaceae, in September, 2-4cm long, long or small long, slightly pointed. It is a deciduous broad-leaved arboreous tree and its origin is Korea. It is distributed in Korea, China, etc. and lives in foothills and valleys.

 Peel this tree and dip it in water, making the water blue. In Gangwon-do, this tree is called Suqinguk (고 水木), and in Oriental medicine, Jinbaekmok (秦 白木). This tree is on the hardest and toughest tree axis. In the past, flails were made of this tree, and still baseball bats and skis are made. Once upon a time, they used to make rice paddies, but they were light and hard to break. Ash trees have been the object of folk belief. Odin, the supreme god of northern Europe, turns into an owl and lives on top of a large ash tree in the woods, where shamans from Europe and Siberia served as a tree. There is a custom left in our country to serve this tree as a sperm.

All eye diseases, including eye redness, conjunctivitis, and trachoma, are often washed with water that has been bark of ash and filtered three or four times with thin gauze. The effect is the same if you cut the ash bark and get the sap to wash your eyes or eye drops. Ash sap clears eyes and aids vision. Consciousness mentioned that it always improves eyesight and prevents all kinds of eye diseases. To treat cataracts or glaucoma, filter the ash sap with bamboo salt, wild honey, or native honey that is more than five years old. If you take eye drops 4 to 7 times a day, you may see unexpected effects.

Chop ash tree branches, boil them for a long time, and steam them with water. Drinking this water together with the poultice is more effective. One thing to note is that alcohol, fish, and tobacco should be prohibited during treatment. Usually in about a week you can see the effect. Ash bark decoction is said to be effective for enteritis and diarrhea and also for bronchitis and asthma.

The components of the dermis known to date are Aesculin, Aesculetin, Coumarin, Ligstroside and Oreuropein. Korean Society, Vol. 50, No. 4, pp 348 ~ 351, Yang Eun-ju, Lee Dong-geun, Lee Jeung-won, Kim Ye-sil, Lim Seon-ha, Song Sok-sik].

In addition, although the above-mentioned dermis is prescribed as a medicinal herb in conventional medicines or related literatures such as Dongbobom, Hyanggyeolbang and light-dose, the prescription is used for the method of differentiating the shape of dermis and the method of manufacturing herbal medicine and liquid solution. It's just a brief comment about it. That is, no opinion on active active ingredients expressing the drug efficacy among the components extracted through the above-described methods could be obtained.

Natural medicines depend on many factors for their quality. In other words, natural medicines may be greatly changed in ingredients or contents depending on the processing methods such as production, cultivation, climate, collection time and drying, numerical value, even in the case of the same herbal medicine. Such instability of natural drugs can be said to be a fundamental cause of difficulty in securing a certain quality, which is an essential requirement of medicines. For this reason, stabilization of quality and establishment of systematic quality management system are essential for natural medicine to function as a modern medicine. Therefore, scientific methods, such as the introduction of physicochemical test-based quality control methods and fingerprint analysis methods using natural indicator components, have been proposed for stabilization (standardization) of natural products.

Indicator material is a component that can represent natural products among the many components contained in natural products. If the characteristics of the raw material are limited to specific indicators, the fractions can be optimized and specified, which can solve the difficulties of standardization and standardization of general plant extracts. In other words, the indicator material is a substance found by research for the standardization of natural products. Currently, there are only 57 kinds of medicinal herbs in the 520 kinds of herbal medicines, and it is difficult to analyze more than two medicines in one research institute each year, and more than 100 billion herbs are needed to investigate all 500 kinds of herbal medicines.

It is difficult to standardize extracts due to the large variation in the content of indicators by region and collection time of dermis, and it is difficult to commercialize because active fractions with excellent anti-inflammatory analgesic, improved blood circulation, and arthritis treatment cannot be reproducibly obtained. It remains a problem to be solved.

In order to solve the problems of the inventors of the present invention as described above, the research efforts to achieve the standardization of the dermis extract according to the specific components and its content control, as a result of suppressing the symptoms of inflammatory changes, anti-inflammatory analgesic action, The standardized dermis extract with excellent effects such as immune cell proliferation inhibitory action, acute inflammation inhibitory, chronic inflammation inhibitory and acute edema inhibition was completed with the present invention.

According to the results of the present invention, it was found that dermis extracts containing a certain amount of caffeine in the dermis extract can be easily and reproducibly obtained, and thus commercialized.

Accordingly, an object of the present invention is to provide a pharmaceutical composition and a food composition for the treatment and prevention of inflammatory diseases comprising the dermis extract as an active ingredient.

In another aspect, the present invention is to provide a dermis extract containing a specific range of caffeine as an indicator material.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

In order to achieve the above object, the present invention is a pharmaceutical composition for the treatment and prevention of inflammatory diseases comprising the dermis (秦 皮, Fraxinus rhynchophylla Hance) extract as an active ingredient, caffeine (Caffeine) as an indicator substance 1.0 to 5.0% by weight dermis (秦 皮, Fraxinus rhynchophylla Hance) Provides a pharmaceutical composition for the treatment and prevention of inflammatory diseases comprising the extract as an active ingredient.

In addition, the present invention, extracting, cooling and filtration of one selected from the group consisting of 5 to 8 times the weight of dermal protozoa weight water, alcohol, alcoholic aqueous solution, the filtrate obtained above is separated into an aqueous solution of alcohol Concentrating under reduced pressure at 50 ~ 90 ℃, and the concentrate obtained in the above, after the alcohol was concentrated under reduced pressure to the filtrate centrifuged at 700 ~ 1,200 rpm to contain 1.0 to 5.0% by weight of caffeine (Caffeine) as an indicator material Preparing an extract; It provides a method for preparing the dermis extract comprising a.

In addition, the present invention by selecting one from the crowd consisting of 5 to 8 times the weight of dermal protozoa weight water, alcohol, aqueous alcohol solution, extraction, cooling and filtration, and then obtained filtrate layered with an aqueous alcohol solution 50 ~ 90 Concentrated under reduced pressure at ℃, the alcohol was added to the obtained concentrate, centrifuged at 700 ~ 1,200 rpm and concentrated under reduced pressure to prevent caffeine (Caffeine) as an indicator material standardized to prevent and treat inflammatory diseases Provide an extract.

In addition, the present invention is a dermal (秦 皮, Fraxinus rhynchophylla Hance) In the inflammatory disease prevention food composition comprising an extract as an active ingredient, caffeine (Caffeine) prepared as an indicator to contain 1.0 to 5.0% by weight dermis (秦 皮, Provides a food composition for preventing inflammatory diseases, including Fraxinus rhynchophylla Hance) extract as an active ingredient. .

According to the present invention, the inflammatory disease is arthritis, edema, immune disease.

According to the present invention, the dermis extract has an excellent anti-inflammatory effect in TPA-induced edema, arachidonic acid-induced edema and carrageenan-induced acute foot edema, anti-inflammatory analgesic effect, arthrase degrading enzyme activity, immune cell proliferation inhibitory effect, The effects of acute inflammation suppression, chronic inflammation suppression and acute edema suppression are excellent, and standardization can be achieved by controlling the content of caffeine as an indicator substance.

Therefore, the dermis extract according to the present invention is extracted from the dermis and prepared by selecting caffeine (Caffeine) as an indicator substance and controlling the content of the indicator substance can be prepared as an inflammatory disease treatment agent and health functional food such as arthritis treatment agent. .

In addition, the composition of the present invention is excellent in safety because it contains a dermis extract that has been used as a herbal medicine for a long time as an active ingredient.

1A shows an HPLC chromatogram of dermis extract.
Figure 1b is a graph showing the LC-Mass results of the dermis extract.
2 is a TPA-induced mouse ear edema. Test of dermis extract.
Figure 3 is arachidonic acid-induced ear edema inhibition test (Arachidonic acid-induced mouse ear edema assay) of the dermis extract.
4 is an acetic acid-induced writhing response in mice.
5 is acute arthritis treatment effect assay of dermis extract.

Hereinafter, the present invention will be described in more detail.

The present invention is standardized and standardized so that the content of caffeine in the dermis extract is included in a certain range to suppress the symptoms of symptoms caused by inflammatory changes such as analgesic suppression, acute inflammation inhibitory, chronic inflammation inhibitory, acute edema inhibition and chronic edema inhibition It relates to a dermal extract that can be excellently expressed and applicable to a medicament useful for the treatment and prevention of inflammatory diseases such as arthritis.

Since the content of the active bioactive substance contained in the original herbal medicine of the dermis extract may vary greatly depending on the place of origin, the harvesting time, the storage period and the storage state, the composition of the herbal composition may not be limited by the weight ratio of the ingredients used. It is more preferable to limit the content. In addition, there is a significant difference in the joint protection bioactivity according to the content of the effective bioactive material.

Therefore, in the present invention, caffeine has not been conventionally proposed as an indicator for obtaining dermis extracts maximizing drug expression. The caffeine activity is involved in contraction of the arterioles and pain in various mechanisms. Caffeine has been widely used to treat headaches by mixing it with pain medications such as aspirin. In addition, caffeine is mixed with ergot alkaloids for the treatment of migraine headaches, which improves the rate of absorption or therapeutic effect [Annals of the Rheumatic Disease 59, 631-635p (2000), Arthritis & Rheumatism, 48 (11): 3055. -3060pp (2003), Lee Young-no, Color Book of Korean Plants, Kyohaksa, 592p (1998), Deokkyun Ahn, Primary Color Book of Korea, Kyohaksa (1998), Lee Woo-cheol, Book of Standard Color Book of Korea, Academy Book (1996), Lee Chang-bok, Korean Plant Book, Hyangmunsa (1979), Jung Tae-hyun, Korea Book of Plants, Shinjisa (1958)].

The dermis extract used in the agent for treating diseases caused by inflammatory changes of the present invention can be selected by extracting the specific active fraction from the extract itself (herbal extract) or dermis extract extracted from the bark of the ash tree. In this case, the compounding ratio is preferably determined by the content of caffeine, which is an indicator substance, regardless of the content of the dermis, which is the original drug. Caffeine, the indicator substance, is preferably contained 1.0 wt% or more in the dermis extract, more preferably 1.0 to 5.0 wt%, even more preferably 1.0 to 3.22 wt%, even more preferably, 3.22 wt% It is most preferable to contain%. If the caffeine content is less than 1.0% by weight, relatively poor results in the treatment of arthritis and the like. There is no particular limitation on the upper limit of the content of caffeine, but if caffeine is contained in excess of a certain level, the effect does not increase anymore, and it is desirable in terms of technical and economic reasons for manufacturing. Tend not to.

Caffeine (Caffeine) selected as the indicator material in the present invention is an important component of the drug for treating diseases caused by inflammatory changes of the present invention, when contained in a certain content range can express a more potent drug effect by expressing a synergistic effect . In addition to caffeine as an active ingredient of the dermis extract obtained in the present invention, it is presumed to exhibit a therapeutic and prophylactic effect of inflammatory diseases as an action of other components. That is, as an active ingredient of the drug obtained in the present invention, the action of other components other than the above components cannot be excluded.

The present invention provides a dermal extract, characterized in that the extraction by selecting one from the crowd consisting of water, alcohol and aqueous alcohol solution.

The extract, 1) extracting, cooling and filtering by selecting one from the group consisting of 5-8 times the weight of dermal protozoa by weight of water, alcohol, and aqueous alcohol solution, 2) separating the filtrate obtained above into an aqueous alcohol solution And then concentrating under reduced pressure at 50 to 90 ° C., and 3) adding the alcohol to the concentrate obtained above, followed by centrifugation at 700 to 1,200 rpm.

The preparation method of the extract is described in more detail as follows.

1) Extracted, cooled and filtered with 6-8 times the amount of water or alcohol solution of washed, dried and cut dermal protozoa, and then added to the residue by adding 5-8 times the amount of water or alcohol solution with water Filtering by re-extracting and then mixing with the previous filtrate and filtering; 2) separating the filtrate obtained in 1) into an aqueous solution of the same amount and then concentrating under reduced pressure at 50 to 90 ° C., and 3) distilling the alcohol recovered upon concentration under reduced pressure of 2) to the concentrate of 2). Concentration under reduced pressure, vacuum drying, grinding and sterilizing the filtrate centrifuged at 700 ~ 1,200 rpm after the addition.

To explain this in detail, washing and drying, adding water or alcohol aqueous solution to the fine dermal protozoa and extracting repeatedly for 2 to 4 hours, 2 to 4 times, slowly cooled at room temperature and filtered by centrifugation, etc. Separate from residue. To the residue is added 6-8 times the amount of water or alcohol aqueous solution of the weight of the mixed herbal medicine, heated and re-extracted, filtered and then mixed with the previous filtrate and filtered. By adding water or an aqueous alcohol solution to the residue as described above, the extraction efficiency can be increased by re-extraction and filtration. At this time, if the amount of the water or alcohol aqueous solution used for extraction is too small, the solubility of the extract is poor, the extraction efficiency is reduced, if the amount is too large, the amount of the aqueous alcohol solution used for layer separation increases, it takes a long time under reduced pressure concentration, etc. May cause economic or handling problems.

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 not economical.

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, in the present invention, the same amount of alcohol as the filtrate. The impurities are purified by separating the aqueous solution using a solvent two to five times to obtain a solvent fraction.

The alcohol is preferably an alcohol having 1 to 6 carbon atoms, more preferably a 30% aqueous ethanol solution. When the amount of the lower alcohol solution is less than the filtrate, fine particles formed by unnecessary components such as fatty acids are formed. Not only is it difficult to separate, but the extraction content of the active ingredient is lowered, which is not efficient.

The alcohol may be used both aliphatic and aromatic alcohols commonly used in the art, preferably, it is preferable to use a lower alcohol having 1 to 6 carbon atoms than aliphatic alcohol.

The layered extract was concentrated under reduced pressure at 50 to 90 ° C. to remove residual solvent. The obtained concentrate was added with alcohol recovered at the time of concentration under reduced pressure, and then the filtrate was centrifuged at 700 to 1,200 rpm. It is cloudy and then concentrated under reduced pressure again.

The concentrate thus obtained is vacuum dried at 0.08 to 0.3 pa at 50 to 90 ° C., and then pulverized and sterilized to 30 to 200 mesh to obtain a powdery dermis extract. The dermis extract has an excellent therapeutic action such as arthritis, and the extract is obtained. Herbal medicines, including, is expected to be useful as a joint protector.

The dermis extract according to the present invention is formulated in a conventional manufacturing method to prepare tablets, capsules, injections, and the like, of which lactose, microcrystalline cellulose, magnesium stearate, etc., which are used as a base when preparing tablets, are combined with the dermis extract. When used in a ratio of 1: 1, a tablet having an activity for treating diseases caused by inflammatory changes such as arthritis can be prepared.

In the preparation of such pharmaceuticals, herbal extracts may be used as such, but they may be mixed with pharmaceutically acceptable carriers, excipients, diluents, etc. to prepare powders, granules, capsules or injections. Is possible. In addition, the dermis 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 condition, diet, administration time, administration method of the patient. , The rate of excretion, the severity of the disease, and the like. Generally, the dermis extract is preferably administered in an amount of about 0.1 to 10 mg per kg of 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.

According to another aspect of the present invention, the present invention provides a food for preventing and treating inflammatory diseases, particularly a health functional food, containing the dermis extract as an active ingredient.

In the present invention, the "health functional food" refers to a food having a bioregulatory function, such as prevention and treatment of diseases, biological defense, immunity, recovery of the disease, inhibition of aging, and should be harmless to the human body when taken in the long term.

In order to use the dermis extract of the present invention for the prevention and treatment of inflammatory diseases as described above, it can be prepared by various methods known in the food science or pharmaceutical field, or by itself or a food acceptable carrier, excipient, diluent, etc. It can be prepared in any food form that can be mixed orally ingested. Preferably in the form of beverages, pills, granules, tablets or capsules.

The dietary supplement of the present invention may further include foodstuffs which are commonly added during food production. For example, when the beverage is prepared, in addition to the plant extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and the like may be further included.

The amount that can be included as an active ingredient of the health functional food according to the present invention may be appropriately selected according to the age, sex, weight, condition, symptoms of the disease of the person who wants to prevent and treat inflammatory diseases, preferably 1 day for adults It is good to include as much as 0.01g to 10.0g, the anti-inflammatory effect can be obtained by eating a health functional food having such a content.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and the scope of the present invention is not to be construed as being limited by these examples.

Example: Preparation of Dermis Extract

After washing with water and drying well, the dermis was added twice with hot water every 2 hours while stirring well with 30% aqueous ethanol solution. The extract was slowly cooled to room temperature and then centrifuged to remove debris, and the filtrates were combined and concentrated under reduced pressure at 50 to 90 ° C.

After the ethanol recovered through the ethanol fraction was added to the concentrate, the concentrate was sufficiently suspended, and the filtrate was centrifuged at 5000 rpm. The filtrate was concentrated under reduced pressure and vacuum dried at 60 ° C. and 0.08 pa. Sterilization was performed through a grinding process. The content of caffeine in the dermis extract extracted and purified by the above method was 3.22% by weight. The dermis extract obtained by the above method was analyzed by LC / Mass, and the instrument manufacturing company was performed by "Hewlett Packerd" and the model by "HP 1100 series LC / MSD" (performed by veterinary department of Chungnam National University). (FIGS. 1A and 1B)

1) Solvent: 20: 1: 79 (Methanol 100%: Acetic acid: Water)

2) Column: C18 reverse phase (Agilent Eclipse XDB-C18 / 1.75㎛, 2.1 * 50mm)

3) flow rate: 1 ml / min

4) Column temperature: 35 ℃

5) Detector: UV 280 nm

As a result, it was confirmed that the same Rt value as caffeine was obtained from the dermis extract obtained by the above method. 1B and 2) of FIG. 1B are peak value results of caffeine, and 3) and 4) are peak value results of dermal extract obtained by the above method, 1) and 3) are LC results, and 2) and 4) are Mass result.

Experimental Example 1: TPA-induced ear edema inhibition test (TPA-induced mouse ear edema.)

TPA-induced mouse edema inhibition test (TPA-induced mouse ear edema.) Was performed on the dermis extract prepared by the above example, and the results are shown in Table 1 and FIG.

Experimental Method 1

Seven-week-old ICR mice were isolated for each experimental group, and then 1 hour after oral administration of 150 mg / Kg of aspirin and 100, 200, 400 mg / Kg of dermal extract, TPA (2.5 μg / 20 μl) was acetone. After dissolving in Aceton, edema was induced in rat ears. In inducing edema, the experimenter completely secured the subject from behind and the second experimenter stimulated the edema causing ear in the ear by using a micro pipet. Four hours later, ear edema was measured for each experimental group. The measurement was carried out using the tibial decay method to induce a precise measurement after culling the specimen.

process Thickness of the ear Inhibition degree of inflammation (%) Non-treatment 0.43 ± 0.026 0 % Aspirin (150 mg / kg) 0.32 ± 0.003 25.58% Dermis Extract (100 mg / kg) 0.33 ± 0.01 23.25% Dermis Extract (200 mg / kg) 0.31 ± 0.01 27.91% Dermis Extract (400 mg / kg) 0.31 ± 0.01 27.91% 1.Data represent the mean of difference in ear thickness (mm) ± SEM (n = 12)
2. No treatment: No treatment of drug after inducing edema

As shown in Table 1, when using the dermis extract prepared according to the present invention, the TPA-induced ear edema inhibitory effect was excellent. Can be.

Experimental Example  2 : Arachidonic acid ( Arachidonic acid ) Induced ear edema suppression test ( Arachidonic acid - induced mouse ear edema assay )

An arachidonic acid-induced mouse ear edema assay was performed on the dermis extract prepared by the above example, and the results are shown in Table 2 and Figure 3 below.

Experimental Method 2

Seven-week-old ICR mice were isolated from each experimental group, and aspirin was dissolved in acetone (2 mg / 20 μl) for 1 hour after oral administration of 150 mg / Kg of aspirin and 100, 200, 400 mg / Kg of dermis extract. Edema caused edema. In inducing edema, the experimenter completely secured the subject from behind and the second experimenter stimulated the edema causing material in the ear by using a micropipette. After 1 hour, ear edema was measured for each natural product. The measurement was carried out using the tibial decay method to induce a precise measurement after culling the specimen.

process Thickness of the ear Inhibition degree of inflammation (%) Non-treatment 0.47 ± 0.01 0 % Aspirin (150 mg / kg) 0.39 ± 0.05 17.02% Dermis Extract (100 mg / kg) 0.35 ± 0.03 25.53% Dermis Extract (200 mg / kg) 0.33 ± 0.01 29.79% Dermis Extract (400 mg / kg) 0.35 ± 0.01 25.53% 1.Data represent the mean of difference in ear thickness (mm) ± SEM (n = 12)
2. No treatment: No treatment of drug after inducing edema

As shown in Table 2, when the dermis extract prepared by the present invention is used, the arachidonic acid-induced ear edema suppression effect is excellent, and in particular, when the dermis extract concentration is 200 mg / kg, the arachidonic acid-induced ear edema inflammation is most preferably It can be seen.

Experimental Example 3: Acetic acid-induced writhing response in mice.

The acetic acid-induced stretching inhibitory test (Acetic acid-induced writhing response in mice) was performed on the dermis extract prepared by the above example, and the results are shown in Table 3 below.

Experimental Method 3

In this experiment, 7-week-old ICR mice were isolated for each herbal candidate by referring to the "Siegmund" test method, and then 1 hour after oral administration of 150 mg / Kg of aspirin, 100, 200, and 400 mg / Kg of dermal extract (0.075). Intraperitoneal administration of 0.1%) (0.1 ml / 10 g) caused inflammation of the organs. After 10 minutes, the mice were stretched for 10 minutes because the swelling causes itching, which can be used to estimate the degree of pain.

process Number of twists
(Writhing number) Pain control degree (%) Non-treatment 17.00 ± 0.00 0 % Aspirin (150 mg / kg) 13.00 ± 0.00 23.53% Dermis Extract (100 mg / kg) 1.50 ± 0.71 91.18% Dermis Extract (200 mg / kg) 9.00 ± 0.82 47.06% Dermis Extract (400 mg / kg) 4.33 ± 0.75 74.51% 1.Data represent the mean of difference in ear thickness (mm) ± SEM (n = 12)
2.No treatment: After treatment of stretching, no treatment treatment

As shown in Table 3, when the dermis extract prepared by the present invention is used, the acetic acid-induced stretching inhibitory effect is excellent, and in particular, when the dermis extract concentration is 400 mg / kg, the acetic acid-induced stretching inhibitory effect is most preferable. .

Experimental Example 4: Acute Arthritis Treatment Effect Test

Acute arthritis therapeutic effect assay was performed on the dermis extract prepared by the above example, and the results are shown in Table 4 and Figure 5 below.

Experimental Method 4

Acute foot swelling was administered to male SD rats (about 200 g) with 150 mg / Kg of aspirin or 1 ml of dermis extract 100, 200, 400 mg / Kg oral, and 100 μl of 1% carrageenan saline solution injected into the left foot. Induced. Edema was measured 4 hours later using a plethysmometer (LE 7500).

process Edema growth rate (%) The degree of suppression of edema (%) Non-treatment 89.36 0 Aspirin (150 mg / kg) 27.04 69.74 Dermis Extract (100 mg / kg) 82.46 7.70 Dermis Extract (200 mg / kg) 65.08 27.17 Dermis Extract (400 mg / kg) 48.08 46.20 1.Data represent the mean of difference in ear thickness (mm) ± SEM (n = 12)
2.No treatment: After treatment of stretching, no treatment treatment

As shown in Table 4, when the dermis extract prepared by the present invention is used, the acute arthritis treatment effect is excellent, and it can be seen that the acute arthritis treatment effect is most preferable when the dermis extract concentration is 400 mg / kg.

Experimental Example 5: Effect on NO

The acute arthritis therapeutic effect assay was performed on the dermis extract prepared by the above example.

Experimental Method 5

After 24 hours of treatment with LPS (1 / ml) in the raw 264.7 cell line, a mouse-derived macrophage, nitric oxide, an inflammation-inducing molecule in the body, was collected by enzymatic and radical chromatography. The effect of dermis extract on the amount of NO) was investigated. NO was determined by measuring the absorbance at OD 540 nm by mixing culture medium 50 and 25 쨉 l of Griess's TS 1 and 2. Absorbance values were measured using an ELISA kit (R & D systems, USA).

dermis
extract
(占 퐂 / ml) 0 0.5 5 50 500 Result Contents Absorbance Suppression rate
(%) Absorbance Suppression rate
(%) Absorbance Suppression rate
(%) Absorbance Suppression rate
(%) Absorbance Suppression rate
(%) shame 0.213 ± 0.087 0 0.204 ± 0.083 4.53 0.200 ± 0.013 6.41 0.170 ± 0.022 20.156 0.122 ± 0.032 42.97

LPS (1 / ml) and dermis extracts (0, 0.5, 5, 50, 500 ㎍ / ml) were treated with raw 264.7 cell line, a mouse-derived macrophage, and cultured in 24 hours As a result of quantifying NO as a triggering molecule, NO production of macrophages by LPS was effectively inhibited in cell lines treated with dermis extract.

Preparation Example 1 Preparation of Tablet

Using the dermis extract prepared according to the embodiment of the present invention, oral administration tablets were prepared using the wet granulation method and the dry granulation method with the following composition.

[Furtherance]

200 mg dermis extract, 10 mg hard silicic anhydride, 2 mg magnesium stearate, 50 mg microcrystalline cellulose, 25 mg sodium starch glycolate, 101 mg lactose, 12 mg povidone, appropriate amount of ethanol anhydride.

Preparation Example 2 Preparation of Ointment

An ointment was prepared using the dermis extract prepared according to the embodiment of the present invention with the following composition.

[Furtherance]

5 g of dermis extract, 20 g of cetyl palmitate, 40 g of cetanol, 40 g of stearyl alcohol, 80 g of myristanisopropyl, 20 g of monostearic acid sorbitan, 60 g of polysorbate, 1 g of paraoxybenzoic acid profile, para 1 g of methyl oxyanate, phosphoric acid and purified water.

Preparation Example 3 Preparation of Injection

Injections were prepared using the dermis extract prepared according to the embodiment of the present invention with the following composition.

[Furtherance]

Dermis extract 100 mg, mannitol 180 mg, dihydrogen phosphate 25 mg, purified water for injection 2974 mg

Preparation Example 4 Preparation of Transdermal Agent

Using the dermis extract prepared according to the embodiment of the present invention, a transdermal agent was prepared in the following composition.

[Furtherance]

0.4 g of dermis extract, 1.3 g of sodium polyacrylate, 3.6 g of glycerin, 0.004 g of aluminum hydroxide, 0.2 g of methyl paraben, 14 g of water.

Preparation Example 5 Preparation of Beverage

Using the dermis extract prepared by the embodiment of the present invention, a beverage was prepared as follows. Purified water is mixed with 200 mg of dermis extract, 75 mg of vitamin C, 4 mg of vitamin B2, 3 mg of vitamin B6, 1,000 mg of dietary fiber, 20000 mg of liquid fructose, 100 mg of emulsifier, and 50 mg of fragrance. The final mixture was clarified by passing the mixed solution through a filter of 2 ~ 3㎛ before sterilization at 90 ~ 93 ℃ for 15 ~ 20 seconds, filled into 50ml bottle and sterilized after 15 ~ 20 minutes at 80 ~ 85 ℃. Completed.

Claims (6)

In the pharmaceutical composition for the treatment and prevention of inflammatory diseases comprising dermis (, Fraxinus rhynchophylla Hance) extract as an active ingredient,
Caffeine (Caffeine) as an indicator material comprising a dermis (1.0, Fraxinus rhynchophylla Hance) extract prepared to contain 1.0 to 5.0% by weight as an active ingredient pharmaceutical composition for the treatment and prevention of inflammatory diseases. The pharmaceutical composition for treating and preventing inflammatory diseases according to claim 1, wherein the inflammatory disease is arthritis, edema, or immune disease. Extracting, cooling and filtering by selecting one from a crowd consisting of 5-8 times the weight of dermal protozoa by weight of water, alcohol and aqueous alcohol solution,
Layering the filtrate obtained above into an aqueous alcohol solution and concentrating under reduced pressure at 50˜90 ° C., and
Preparing an extract containing 1.0 to 5.0% by weight of caffeine as an indicator material by concentrating the filtrate obtained by adding alcohol to the concentrate obtained above, and centrifuging the filtrate at 700 to 1,200 rpm under reduced pressure; Method for producing a dermis extract comprising a. Select one from a crowd consisting of 5-8 times the weight of dermal protozoa, water, alcohol, and aqueous alcohol solution, extract, cool, and filter. The filtrate was separated with an aqueous solution of alcohol and concentrated under reduced pressure at 50 ~ 90 ℃. In addition, alcohol was added to the obtained concentrate, and the filtrate was centrifuged at 700 to 1,200 rpm. The filtrate was concentrated under reduced pressure to extract 1.0 to 5.0% by weight of caffeine as an indicator. . In the food composition for preventing inflammatory diseases comprising dermis (피, Fraxinus rhynchophylla Hance) extract as an active ingredient,
Caffeine (Caffeine) as an indicator material containing a dermis (秦 皮, Fraxinus rhynchophylla Hance) extract prepared to contain 1.0 to 5.0% by weight as an active ingredient food composition for preventing inflammatory diseases. The food composition of claim 5, wherein the inflammatory disease is arthritis, edema, or immune disease.

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