KR100779026B1 - Osmanthus heterophylla extracts compositions for treating or preventing inflammatory diseases - Google Patents

Abstract

The present invention relates to a bone extract useful for the treatment and prevention of inflammatory diseases, more specifically, standardized and standardized to include a certain amount of caffeine in the extract of the bone, pain suppression, acute inflammation inhibitory, chronic inflammation inhibitory, acute The present invention relates to a bulbar extract that can be used as a medicament useful for treating and preventing diseases caused by inflammatory changes such as arthritis due to excellent expression of inhibitory effects caused by inflammatory changes such as edema suppression and chronic edema suppression.

Osteotomy, Arthritis, Acute Inflammation, Acute Edema, Caffein

Description

Osmanthus heterophylla extracts compositions for treating or preventing inflammatory diseases

       Figure 1 shows the HPLC chromatogram of the gogol extract according to the present invention.

The present invention relates to a bone extract useful for the treatment and prevention of inflammatory diseases, more specifically, standardized and standardized to include a certain amount of caffeine in the extract of the bone, pain suppression, acute inflammation inhibitory, chronic inflammation inhibitory, acute The present invention relates to a bulbar extract that can be used as a medicament useful for treating and preventing diseases caused by inflammatory changes such as arthritis due to excellent expression of inhibitory effects caused by inflammatory changes such as edema suppression and chronic edema suppression.

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 arthritis (보 性): Usually caused by trauma (外傷), the cause is unknown, usually occurs in only one joint. ② serous fibrosis (奬 液 纖維素 性) arthritis: acute arthritis occurs during the arthritis, turbid effusion in the joint cavity (液 出 강) 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 in the open window (開放 創) or gonorrhea, typhoid fever, scarlet fever, sepsis (敗血症) has multiple symptoms. Infants 1-2 months of age are severely damaged bones, causing dislocations that cannot be treated. 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 gout in middle-aged men due to the metabolic disorders of many uric acid (痛風 性). ② Multiple arthritis: Chronic arthritis is caused by a lot of arthritis in acute serous arthritis, (Transition), tuberculosis, syphilis, gonorrhea in the course of multiple, and may be one of the sepsis. In addition, it also includes arthritis called Still's disease. ③ deformable osteoarthritis: bone or joint aging or trauma is the cause. ④ hemophilia (血友病 性) arthritis: when hemophilia is due to 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, Osgolthus ( Osmanthus heterophylla ), also known as a hwalyeob, lives mainly on the seashore and grows to a light pale gray-brown color. Leaves are opposite, oval or oval upside down, thick, 3-5 cm long, 2-3 cm wide. The leaf front is dark green and glossy, and the back is light green or yellowish green. Young leaves have this shape of projections, and petioles are 7-12 mm long. Flowers are single flowers, bloom in November, white, clustered in axilla, small petioles 5-12 mm. The calyx and corolla are divided into four ends and the stamens are two. The fragment of the calyx is flat with an egg-shaped triangle. Fruits are nucleus, oval, and ripe to blackish purple, mainly distributed on Jeju Island.

The traditional Chinese medicine and related literature, such as Dongbobogam, Hyanggyeolbang and light-dose, have been prescribed as the unilateral herbal medicine, but these prescriptions are a simple method of differentiating the shape of the bone and manufacturing methods of herbal medicine and sugar solution. It was only a mention. In other words, opinions regarding the active ingredients expressing the efficacy of the components extracted through the above-described method could not be obtained.

In addition, it is difficult to standardize extracts because of the large variation in the content of the indicators by the place of origin and harvest, and it is difficult to commercialize because the active fractions with excellent anti-inflammatory analgesic, improved blood circulation, and arthritis treatment cannot be reproducibly obtained. The fact remains a problem to be solved.

The inventors of the present invention solve the problems as described above, suppression of symptoms caused by inflammatory changes, anti-inflammatory analgesic action, platelet aggregation inhibitory action, articular tissue degrading enzyme inhibitory action, immune cell proliferation inhibitory action, acute inflammation inhibitory, acute Efforts have been made to achieve standardization of goji extract by controlling certain components and its content with excellent effects such as edema suppression.

According to the results of the present invention, it is possible to obtain commercially available gourd extract containing a certain amount of the active fraction which is considered to be excellent in inhibiting symptoms of the inflammatory disease among the goul extracts. I learned.

Accordingly, an object of the present invention is to provide a bone extract useful for the treatment and prevention of inflammatory diseases.

In another aspect, the present invention is to provide a bone extract extract containing a specific range of caffeine is determined as an indicator of the above-mentioned bulbar extract.

According to the present invention, an osteoclast containing 0.5 to 5.0% by weight of caffeine as an indicator material ( Osmanthus heterophylla ) is characterized in that the extract for the treatment and prevention of inflammatory diseases, characterized in that the 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 extract of the bulbus is suppressed by the inflammatory changes such as analgesic suppression, acute inflammation inhibitory, chronic inflammation inhibitory, acute edema inhibition and chronic edema inhibition Is excellently expressed and relates to a bone extract that can be applied to a medicament useful for treating diseases caused by inflammatory changes such as arthritis.

Since the content of the active bioactive substances contained in the original herbal medicines can vary greatly depending on the place of origin, harvesting time, storage period, and storage conditions, the content of a specific active substance in the composition of each herbal composition is more limited than the weight ratio of the ingredients used. It is more preferable to limit.

Therefore, in the present invention, caffeine was selected as an indicator for obtaining a bone extract to maximize the expression of drug efficacy. 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)].

Gugol bone 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 from the gugol extract. In this case, the mixing ratio is preferably determined by the content of caffeine, which is an indicator material, regardless of the original content of the bone.

That is, in the case of a drug for treating diseases caused by inflammatory changes of the present invention, when the content of caffeine, which is the indicator substance, is contained in 0.5 to 5.0% by weight of the bulbar extract used in the drug, a desired drug can be obtained. At this time, when the content of caffeine is less than 0.5% by weight, relatively poor results in the treatment of arthritis and the like are obtained. In addition, in the present invention, there is no particular limitation on the upper limit of the caffeine content, but if it contains more than a certain level, the effect does not increase any more, but also tends to be undesirable in terms of production, technical and economic. . Preferably, the content of caffeine is 0.5 to 5.0% by weight, and most preferably about 2.7% by weight of the bone extract.

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 by expressing a synergistic effect. In addition, as an active ingredient of the agent for treating diseases caused by the inflammatory change of the present invention obtained in the present invention, the action of other components other than the above components cannot be excluded.

The bone extract used in the present invention for the treatment of inflammatory diseases is a powder containing 0.5 to 5.0% by weight of caffeine.

1) Extract, cool and filter with 5-8 times water, alcohol or alcohol aqueous solution of the bulbar crude drug, and add 2-5 times water, alcohol or alcohol aqueous solution of the mixed crude drug weight to the residue and warm it again. Extraction and filtration, followed by filtration with the previous filtrate,

2) concentrating the filtrate obtained in 1) at 60-80 ° C. under reduced pressure,

3) adding to the concentrate obtained in step 2) the alcohol recovered during the concentration under reduced pressure of 2), followed by concentration under reduced pressure, vacuum drying, grinding and sterilizing the filtrate obtained by centrifugal filtration.

If this is described in detail, water or alcohol solution is added to the nasolabial protozoa and extracted 2 to 5 hours, 2 to 3 times, and after cooling slowly at room temperature, it is filtered by centrifugation or the like to separate from the residue. To the residue is added 5-8 times the amount of water or alcohol aqueous solution of the weight of the mixed herbal medicines, 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 water, alcohol or an aqueous alcohol solution over the first and second stages is filtered and concentrated, and then purified impurities, such as unnecessary proteins, polysaccharides and fatty acids contained in the filtrate, in the present invention is the same amount as the filtrate The impurities are purified by adding an aqueous alcohol solution 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.

As the alcohol, both aliphatic and aromatic alcohols generally used in the art may be used, and preferably, lower alcohols having 1 to 6 carbon atoms are more preferable than aliphatic alcohols.

The filtrate is concentrated under reduced pressure at 60 to 80 ° C. to remove the remaining solvent. The alcohol obtained at the time of concentration under reduced pressure is added to the concentrate thus obtained, and the filtrate obtained by centrifugal filtration is again concentrated under reduced pressure.

The concentrate thus obtained is vacuum dried at 0.08 to 0.3 pa at 60 to 80 ° C., and then pulverized and sterilized to 30 to 200 mesh to obtain a powdery bulbus extract. These extracts have excellent therapeutic effects such as arthritis. Herbal medicines, including, is expected to be useful as a joint protector.

Tablets, capsules, injections, etc. are prepared by formulating the bone extract according to the present invention in a conventional manufacturing method, among which the lactose, microcrystalline cellulose, magnesium stearate, etc., which are used as a base for the preparation of tablets, are combined with the bone extract 1 When used in a ratio of 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 bone extract according to the present invention has been used for edible and medicinal since ancient times, there is no particular restriction on the dosage, body absorption, weight, age, sex, health status, diet, administration time, administration method of the patient , The rate of excretion, the severity of the disease, and the like. Generally, it is preferable to administer about 0.1-10 mg / kg body weight extract.

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, which are not intended to limit the present invention.

Example: Preparation of Gogol Extract

After washing with water and drying well, the fine bone was added with 30% ethanol solution in the same amount and stirred well every two hours while stirring well. 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 60 to 80 ° 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 golgol extract extracted and purified by the above method was 2.7% by weight. In addition, HPLC analysis of the gogol extract obtained by the above method under the following conditions.

1) Solvent:

 A: 0.1% acetic acid solution

 B: acetonitrile

2) Column: C ₁₈ column (4.6 mm × 250 mm) or equivalent or more

3) Flow rate: 1.0 ml / min

4) Column temperature: 20 ℃

5) Detector: UV 280 nm

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

TPA-induced ear edema inhibition test (TPA-induced mouse ear edema.) Was performed on the extracts of the bones prepared by the above examples, and the results are shown in Table 1 below.

Experimental Method 1

Seven-week-old ICR mice were isolated from each experimental group, followed by oral administration of 200 mg / Kg of aspirin and 100, 200, 400 mg / Kg of bone extracts. TPA (2.5 μg / 20 μl) 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 Inflammation-induced degree (%) Negative control 0.295 ± 0.058 39% Positive control 0.765 ± 0.010 100% aspirin 0.381 ± 0.010 49% Gol bone (100 mg / kg-caffeine content 0.5 wt%) 0.604 ± 0.019 79% Gugol (200 mg / kg-caffeine content 1.0 wt%) 0.551 ± 0.014 72% Gugol (400 mg / kg-caffeine content 2.0 wt%) 0.553 ± 0.024 72% Gut bone (0.25% by weight of 100 mg / kg-caffeine content) 0.704 ± 0.012 92% 1. Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12) 2. Negative control: Experimental group that did not cause edema 3. Positive control: Drug-free experimental group after edema induction

 As shown in Table 1, the TPA-induced ear edema inhibitory effect is excellent when using the bone extract prepared according to the present invention, and particularly, the concentration of TPA-induced ear edema inflammation is most preferred when the extract is 200 mg / kg. Can be.

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

Arachidonic acid-induced mouse ear edema assay was performed on the bulbar extract prepared by the above example, and the results are shown in Table 2 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 200 mg / Kg of aspirin and 100, 200, 400 mg / Kg of bone 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 using a micro pipette. 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 Inflammation-induced degree (%) Negative control 0.303 ± 0.004 40% Positive control 0.755 ± 0.008 100% aspirin 0.349 ± 0.008 46% Gol bone (100 mg / kg-caffeine content 0.5 wt%) 0.536 ± 0.011 71% Gugol (200 mg / kg-caffeine content 1.0 wt%) 0.516 ± 0.010 68% Gugol (400 mg / kg-caffeine content 2.0 wt%) 0.528 ± 0.012 70% Gut bone (0.25% by weight of 100 mg / kg-caffeine content) 0.687 ± 0.011 91% 1. Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12) 2. Negative control: Experimental group that did not cause edema 3. Positive control: Drug-free experimental group after edema induction

 As shown in Table 2, the use of the kogol extract prepared by the present invention is excellent in inhibiting arachidonic acid-induced ear edema. 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 bulbar 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 experimental method of "Siegmund", and then 1 hour after oral administration of 200 mg / Kg of aspirin and 100, 200, 400 mg / Kg of bone extract, 0.7%) intraperitoneal administration (0.1 ml / 10 g) caused organ inflammation. After 10 minutes, the mice were stretched for 10 minutes because of the itching caused by edema, which can be used to estimate the degree of edema.

process Writhing number Inflammation-induced degree (%) Positive control 52.00 ± 0.92 100% aspirin 10.17 ± 0.57 20% Gol bone (100 mg / kg-caffeine content 0.5 wt%) 3.17 ± 0.22 61% Gugol (200 mg / kg-caffeine content 1.0 wt%) 27.17 ± 1.33 52% Gugol (400 mg / kg-caffeine content 2.0 wt%) 28.10 ± 0.31 54% Gut bone (0.25% by weight of 100 mg / kg-caffeine content) 45.76 ± 0.55 88% 1.Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12) 2. Positive control

As shown in Table 3, the use of the extract from the bone bone prepared by the present invention is excellent in inhibiting arachidonic acid-induced stretching, and particularly, the concentration of arachidonic acid-induced stretching is most preferred in the case of the concentration of 200 mg / kg. Can be.

Experimental Example 4: Acute Arthritis Treatment Effect Test

The acute arthritis treatment effect assay was performed on the bulbar extract prepared by the above example, and the results are shown in Table 4 below.

Experimental Method 4

200 mg / Kg of S company J, pre-prepared to male SD rats (about 200 g), oral administration of 100 ml, 100, 200, 400 mg / Kg of bone ball extracts were injected orally, and 100 µl of 1% carrageenan saline was injected into the left foot. Acute foot edema was induced. Edema was measured using a plethysmometer (LE 7500) after 3 hours.

process Inflammation-induced degree (%) Negative control 0 % Positive control 89% S company J product 70% Gol bone (100 mg / kg-caffeine content 0.5 wt%) 82% Gugol (200 mg / kg-caffeine content 1.0 wt%) 78% Gugol (400 mg / kg-caffeine content 2.0 wt%) 78% Gut bone (0.25% by weight of 100 mg / kg-caffeine content) 81% 1. Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12) 2. Negative control: Experimental group that did not cause edema 3. Positive control: Drug-free experimental group after edema induction

As shown in Table 4, the use of the osteoblast extract prepared according to the present invention is excellent in treating acute arthritis, and particularly in the case of the extract of the osteoblast, 200 mg / kg.

Experimental Example 5: Effect on NO and Prostaglandin E2 Production

Acute arthritis therapeutic effect assay was performed on the bulbar extract prepared by the above example.

Experimental Method 5

After treatment with LPS (1 ㎍ / ml) in raw 264.7 cell line, a mouse-derived macrophage, the culture solution was collected 24 hours later, and nitric oxide, an inflammation-inducing molecule in the body, was analyzed by enzyme and radical chromatography. , NO) and the effect of bulbar extract on the production of prostaglandin E2 (PGE2). In the measurement of NO, 50 μl of the culture solution was taken and 25 μl of Greases solution was mixed, and the absorbance at 540 nm was measured. PGE2 was measured using an ELISA kit (R & D systems, USA).

After treatment with LPS (1 ㎍ / ml) and bulbar extract (25 ㎍ / ml) in Raw 264.7 cell line, which is a mouse-derived macrophage, the culture medium was collected 24 hours later, and NO was determined by using enzyme method. As a result, when medicinal herbs and LPS were not treated, low concentrations of nitrite of less than 10 μM were detected, but about 40 μM of nitrite could be detected from the culture of cell lines treated with LPS alone for 24 hours. This means that the production of NO in the cells was increased by the expression of immune-related proteins such as NO _S by LPS. In the cell line treated with the bulbus extract, the concentration was detected at a concentration of 5-25 μM that was significantly suppressed than 40 μM. NO production of macrophages was suppressed.

Experimental Example 6 Comparison of the Expression of Inflammation-Related Genes by Real Time-PCR

Comparison of the expression levels of inflammation-related genes with respect to the bulbar extract prepared by the above example was compared to COX2 (Cyclooxygenase, iNOS (nitrogen Oxide Synthetase), mRNA expression changes in real-time PCR (Rotar gene 3000, Corbett) and Western blotting ( Western blotting).

[Test Method 6-1] COX2 (Cyclooxygenase 2), iNOS (nitrogen Oxide Synthetase) Gene Expression Inhibition Experiment

To investigate the anti-inflammatory effects of bone extracts and their correlation with inflammation-associated molecules, we investigated the effects of bone extracts on the expression of COX2 (Cyclooxygenase 2) and nitrogen Oxide Synthetase (iNOS) genes and proteins in real-time (Rotar gene 3000, Corbett). And western blotting.

Experimental Method 6-2] Measurement of mRNA Expression Change

LPS (lipopolysaccharide) (1 ㎍ / ml) and the bone extract were treated with RNA by Qiagen RNasy kit from Raw 264.7 cells incubated for 8 hours, and cDNA was synthesized and quantitatively analyzed by real-time PCR. Real-time RT-PCR was performed by real-time PCR using the SYBER GREEN or Taqman method with the cDNA template.

[Experimental Method 6-3] Western Blotting

After 24 hours of LPS and bulbar extracts, the protein isolated from the cells was electrophoresed on a 10% SDS polyacrylamide gel and the protein on the gel was transferred to a PVDF membrane under an appropriate voltage. Membranes were blocked for 2 hours at room temperature with tris buffer (25 mM Tris, pH 7.6, 192 mM NaCl) containing 10% whey milk and 0.5% BSA. Antibodies for each factor were diluted in TBST containing 3% bovine serum albumin in an appropriate ratio and added to the membrane, and then labeled with shaking for 1 hour at room temperature. The membrane was washed three times with PBST, immersed in an appropriate secondary antibody solution, reacted at room temperature for 1 hour, and washed. ECL kit was used to confirm the photosensitive X-ray film for a suitable time.

Gene expression of immune-related molecules of macrophages was induced by LPS treatment, and the effects of medicinal herbs on their expression levels were compared and quantified by real-time PCR. The results were as follows.

That is, the induction effect of LPS was confirmed by comparing the expression levels of the LPS treated group and the untreated group. MRNA expression of COX-2 increased about 66-fold after 8 hours of LPS treatment.

In addition, mRNA expression level of iNOS increased more than 12 times after 8 hours of LPS treatment, and the treatment of goose extract showed 49% expression inhibition effect. After 24 hours of LPS and drug treatment, 50 μg of protein isolated from cells was quantified and electrophoresed with 10% SDS polyacrylamide gel, followed by Western blotting as described above. Inflammation-related proteins COX-2 and iNOS showed the highest expression after 24 hours of LPS treatment, but did not express both proteins without LPS treatment.

Experimental Example 7: Effect on platelet aggregation reaction

Experimental Method 7

To detect the antiplatelet action of the bulbar extract, blood was collected from the rabbit's abdominal artery using a syringe pre-filled with sodium citrate solution and centrifuged at 200 g for 10 minutes, followed by platelet-rich plasma (PRP). Re-centrifuged some PRP for 10 minutes at 2.000 g to obtain platelet-poor plasma (PPP), the supernatant except for precipitation, and the PRP to be used for platelet aggregation experiments with PPP. Diluted to a concentration of 10 ⁸ cell / ml. Platelet aggregation was induced by collagen, and the degree of aggregation was measured according to the turbido-metric method using a platelet aggregometer. The concentrations of the samples used in the search were 500, 300 and 100 µg / ml.

Platelet-rich plasma (PRP) obtained from rabbit blood was prepared to have a platelet count of 2 × 10 ⁸ cells / ml, and then platelet aggregation was induced with collagen, and the degree of aggregation was measured using a platelet aggregometer. According to the turbido-metric method, the following results were obtained.

The anti-platelet aggregation inhibitory effect was about 20% in the experimental group treated with 300 ㎍ / ml concentration when treated with the extract of old bone.

Preparation Example 1 Preparation of Tablet

Tablets for oral administration were prepared using the wet granules method and the dry granules method with the following composition using the bulbar extract prepared according to the embodiment of the present invention.

[Furtherance]

Gogol bone extract 200 mg, hard silicic acid anhydrous 10 mg, magnesium stearate 2 mg, microcrystalline cellulose 50 mg, starch glycolate 25 mg, lactose 101 mg, povidone 12 mg, ethanol anhydride.

Preparation Example 2 Preparation of Ointment

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

[Furtherance]

Gogol bone 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.

Preparation Example 3 Preparation of Injection

Injections were prepared with the following composition by using the extract from the bulbar bone prepared according to the embodiment of the present invention.

[Furtherance]

Gugol bone extract 100 mg, mannitol 180 mg, sodium dihydrogen phosphate 25 mg, purified water for injection 2974 mg

Preparation Example 4 Preparation of Transdermal Agent

The percutaneous agent was prepared by the following composition using the bone extract prepared according to the embodiment of the present invention.

[Furtherance]

0.4 g of gogol 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 ml of an acrylic adhesive solution.

As described above, according to the present invention, as an inhibitor of inflammatory changes, anti-inflammatory analgesic action, platelet aggregation inhibitory action, articular tissue degrading enzyme action, immune cell proliferation inhibitory action, acute inflammation inhibitory effect, chronic inflammation inhibitory effect and Standardization of the bone extract according to the control of the specific component and its content with excellent effects such as acute edema suppression can be achieved.

In addition, the bone extract according to the present invention is prepared by extracting from the bone can be selected for caffeine as an indicator material can be prepared for the treatment of arthritis therapeutic activity by controlling the content of the indicator material.

Claims (3)

An agent for the treatment and prevention of arthritis, edema, comprising as an active ingredient an extract of Osmanthus heterophylla containing 0.5 to 5.0% by weight of caffeine as an indicator. According to claim 1, wherein the gogol extract is 1) extracting, cooling, and filtering with 5 to 8 times the amount of water, a C1-6 alcohol, or an aqueous C1-6 alcohol solution; 2) concentrating the filtrate obtained above under reduced pressure to 60-80 ° C .; And 3) concentrating the vacuum filtrate obtained by adding alcohol having 1 to 6 carbon atoms and centrifugal filtration to the concentrate obtained above, vacuum drying, pulverizing and sterilizing; It is prepared, including, the drug characterized in that the powder containing 0.5 to 5.0% by weight caffeine. delete

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