KR20070088940A - Amaranthaceae extracts compositions for treating or preventing inflammatory diseases - Google Patents

Abstract

Provided is a medicine comprising an extract of Amaranthaceae having constant amount of succinic acid which shows excellent inhibitory effect on various symptoms such as pain inhibition, acute inflammation inhibition, chronic inflammation inhibition, acute edema inhibition and chronic edema inhibition so as to be usefully used for preventing and treating inflammatory diseases. The medicine for preventing and treating inflammatory diseases comprises an extract of Amaranthaceae which contains 0.5-5.0 wt.% of succinic acid as an index material. The extract is obtained by the method comprising the steps of: (a) extracting Amaranthaceae with water, alcohol, or an aqueous solution of alcohol, cooling, and filtering it; (b) after layer-separating the filtrate obtained from the step (a) using an aqueous solution of alcohol, concentrating it at a temperature of 60-80 deg.C under reduced pressure; and (c) after adding alcohol to the concentrate and centrifuging it with the speed of 500-1,000 rpm, concentrating it under reduced pressure, drying it under vacuum, pulverizing and sterilizing it.

Description

Amaranthaceae extracts compositions for treating or preventing inflammatory diseases

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

The present invention relates to a medicament useful for the treatment and prevention of inflammatory diseases containing dew extract, more specifically, standardized and standardized so as to include a certain range of succinic acid in the extract of dew, Inhibitors of inflammatory changes such as acute inflammation suppression, chronic inflammation suppression, acute edema suppression and chronic edema suppression have excellent expression inhibitory effect and are useful for the treatment and prevention of inflammatory diseases containing arthritis extract 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 arthritis (奬 液 性) arthritis: usually caused by trauma, the cause of unknown, and 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 pus to burst and pus enters the joints, which is called secondary arthritis.

Chronic arthritis is classified as follows: ① Specific Inflammation: Tuberculosis, syphilis, or post-menopausal arthritis due to many metabolic disorders of uric acid in middle-aged men. 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, Amaranthaceae is a herbal medicine that is dried in the sun after removing roots from the roots of Achyranthes japonica, a plant and amaranth, which are distributed throughout Korea, Japan, and South Yellow River in China. Yellowish brown stick-shaped or slightly curved, 15-90 cm long, 3-7 cm in diameter, cylindrical, with many longitudinal wrinkles and marks of side roots. The plane is grayish white or light brown, and the neck in the center is yellowish white and the quality is hard but brittle, almost no smell, the taste is slightly sweet, mucus, and the cross section of the skin and the neck is a clear formation layer. Are distinguished. At the center of the neck, there is a small protozoa, surrounding it, and the concentric annulus is arranged outward. The flow cell layer contains the matter of calcium hydroxide, and starch granules are not seen.

Its pharmacological action is known to be effective in diuretic, painful, arthritis, etc. The main ingredients are composed of saponin, oleanoic glycoside, and succinic acid. (The Korean Journal of Herbal Sciences), Vol. 13, No. 1, pp 95-118, Shin Min-kyo, Song Ho-jun, Byun-ho, Son In-kyung, No Byung-kyu].

Although traditional medicines and related literatures such as Dongbobom, Hyanggyeolbang and light-dose are prescribed for the above-mentioned dew as a single herbal medicine, such prescription is a simple method of differentiating the shape of the dew and the method of manufacturing herbal medicine and liquid 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 the extract because it has a large content deviation of the indicator component by the place of origin and the harvesting time, and commercialization is difficult because the active fraction with excellent activity such as anti-inflammatory pain, improved blood circulation, and arthritis treatment cannot be reproducibly obtained. The fact remains a problem to be solved.

Therefore, the inventors of the present invention solve the problems described above as an inhibitory effect of inflammatory changes, anti-inflammatory analgesic action, platelet aggregation inhibitory action, articular tissue degrading enzyme inhibitory action, immune cell proliferation inhibitory action, acute inflammation inhibitory, chronic Research efforts have been made to achieve standardization of hyssop extracts according to specific ingredients with excellent effects such as inflammation suppression and acute edema suppression and its content control.

According to the results of the present invention, it is possible to commercialize the dew extract containing a certain amount of the active fraction, which is considered to be excellent in the inhibitory effect of the above-mentioned inflammatory disease among the dew extract. I learned.

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

It is another object of the present invention to provide a dew drop extract containing a specific range of succinic acid, which is considered to be an indicator substance, which is an active fraction having an excellent inhibitory effect of inflammatory changes among the dew drop extracts. .

The present invention is characterized by a drug for the treatment and prevention of inflammatory diseases, which is an extract of Amaranthaceae containing 0.5 to 5.0% by weight of succinic acid as an indicator.

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

The present invention is standardized and standardized to include a certain amount of succinic acid in the extract of dew drops to suppress the pain caused by inflammatory changes, such as analgesic suppression, acute inflammatory inhibitors, chronic inflammatory inhibitors, acute edema inhibitors and chronic edema inhibitors The present invention relates to a dew extract which can be applied to a medicament useful for the treatment and prevention of inflammatory diseases such as arthritis due to its excellent expression of symptoms.

The agent for treating diseases caused by inflammatory changes of the present invention includes the extract of dew drops as an active ingredient, and it is preferable to use the extract containing 0.5 to 5.0% by weight of succinic acid as an indicator substance.

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, the succinic acid is selected as an indicator for obtaining the extract of the dew drops to maximize the expression of the drug. The succinic acid is a chemical core precursor composed of four carbons and two carboxyl groups, and is widely used in the chemical, food, and pharmaceutical industries.

The dew extract used in the medicament for the treatment and prevention of inflammatory diseases of the present invention can be selectively used by extracting the specific active fraction from the extract itself (herbal extract) or the extract of the dew. In this case, the mixing ratio is preferably determined by the content of succinic acid, which is an indicator substance, regardless of the original content of the dew.

That is, in the case of the drug for treating diseases caused by the inflammatory change of the present invention, when the content of succinic acid, which is the indicator substance, is contained in the extract of 0.5% to 5.0% by weight of the dew extract used in the medicine, the desired effect could be obtained. . At this time, when the content of the succinic acid (succinic acid) is less than 0.5% by weight it is relatively poor in the treatment of arthritis. In addition, in the present invention, there is no particular limitation on the upper limit of the succinic acid content, but if it is contained in excess of a certain level, the effect does not increase any more, and it is not preferable in terms of technical and economical manufacturing. Therefore, it is most preferable to use about 3.0% by weight.

Succinic acid (succinic acid) selected as the indicator material in the present invention is an important component of a drug for treating diseases caused by inflammatory changes of the present invention, when expressed in a certain content range can express a more potent drug effect by expressing a synergistic effect have. In addition, as an active ingredient of the drug 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.

Brief description of the preparation method of hyssop extract effective for the treatment of diseases caused by inflammatory changes of the present invention according to the present invention.

1) extracting, cooling and filtering with 5-8 times the weight of wort protozoa by water, an alcohol or an aqueous alcohol solution;

2) separating the filtrate obtained above into an aqueous alcohol solution and concentrating under reduced pressure at 60 to 80 ° C., and

3) Concentration of the filtrate obtained by adding alcohol, centrifugation at 500 ~ 1,000 rpm and then concentrated under reduced pressure, vacuum drying, grinding and sterilization.

To explain this in detail, water, alcohol or an aqueous alcohol solution is added to the raw crude medicine, and extracted repeatedly for 2 to 3 hours and 2 to 5 times. After cooling slowly at room temperature, the mixture is filtered by centrifugation or the like to separate 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 water, alcohol or alcohol aqueous solution used for extraction is too small, the solubility of the extract is poor, the extraction efficiency is lowered. If the amount is too high, the amount of alcohol aqueous solution used for layer separation increases, and the decompression concentration is long It may not be economical, such as jamming or handling problems.

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.

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 with 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 layered extract was concentrated under reduced pressure at 60 to 80 ° 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 500 to 100 rpm. It is cloudy and concentrated under reduced pressure again.

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 dew drop extract. The dew drop extract has an excellent therapeutic action such as arthritis. Herbal medicines, including, is expected to be useful as a joint protector.

To prepare a tablet, capsules injections, etc. by formulating the extract of the dew drops according to the conventional method, among them, a combination of lactose, microcrystalline cellulose, magnesium stearate, etc., which is used as a base for the preparation of tablets, and the 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 dew extract according to the present invention has been used for food and medicinal use since ancient times, and there is no particular limitation 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. In general, it is preferable to administer about 0.1 to 10 mg of 1 g of body weight extract per 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 by the examples.

Example 1 Preparation of Emulsion Extract

After washing with water and drying well, about 30% aqueous ethanol solution was added thereto, followed by stirring with hot water twice every 2 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 then the filtrate was centrifuged at 1000 rpm and concentrated under reduced pressure, followed by vacuum drying at 60 ° C. and 0.08 pa. Sterilization was performed through a grinding process. The content of succinic acid was 0.5-5.0 wt% in the extract of dew drops purified by the above method. The dew extract obtained by the above method was also analyzed by HPLC under the following conditions.

1) Developing solvent (Eluent): 0.005 N sulfonic acid

2) Column: Rezex ROA-organic acid, 300 × 7.8 mm I.D., 5㎛

3) Flow rate: 0.5 ml / min

4) Column temperature: 55 ℃

5) Detector: UV 210 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 dew extract prepared in Example 1, and the results are shown in Table 1 below.

Experimental Method 1

Seven-week-old ICR mice were isolated from each experimental group, and then 1 hour after oral administration of 200 mg / Kg of aspirin and 100, 200, 400 mg / Kg of dew extract, acetone of 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. When the measurement was carried out using the tibial decay method, the test subjects were culled and accurate measurements were induced.

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% Whipped (100 mg / kg-succinic acid content 0.5% by weight) 0.619 ± 0.027 81% Whipped (200 mg / kg-succinic acid content 1.0 wt%) 0.597 ± 0.014 78% Whipped (400 mg / kg-succinic acid content 2.0 wt%) 0.596 ± 0.012 78% Whipped (50 mg / kg-Succinic acid 0.25 wt%) 0.760 ± 0.011 99% 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, when using the dew extract prepared according to the present invention, the TPA-induced ear edema inhibitory effect is excellent, and particularly, the concentration of 200 mg / kg of dew drop extract shows that TPA-induced ear edema inflammation is most preferable. 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 extract of E. coli, prepared by Example 1, 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 dew 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% Whipped (100 mg / kg-succinic acid content 0.5% by weight) 0.596 ± 0.015 79% Whipped (200 mg / kg-succinic acid content 1.0 wt%) 0.562 ± 0.012 74% Whipped (400 mg / kg-succinic acid content 2.0 wt%) 0.566 ± 0.014 75% Whipped (50 mg / kg-Succinic acid 0.25 wt%) 0.752 ± 0.013 99% 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, when the dew extract prepared according to the present invention is used, the arachidonic acid-induced ear edema suppression effect is excellent. It can be seen.

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

The arachidonic acid-induced stretching inhibitory test (Arachidonic acid-induced writhing response in mice) on the extract prepared from Example 1 was carried out, 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 1 hour after oral administration of 200 mg / Kg of aspirin, 100, 200, and 400 mg / Kg of aloe extract, respectively. (0.075%) of 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% Whipped (100 mg / kg-succinic acid content 0.5% by weight) 24.20 ± 0.96 61% Whipped (200 mg / kg-succinic acid content 1.0 wt%) 20.50 ± 0.86 39% Whipped (400 mg / kg-succinic acid content 2.0 wt%) 21.42 ± 0.88 41.2% Whipped (50 mg / kg-succinic acid content 0.25% by weight) 51.00 ± 0.90 99% 1.Data represent the mean of difference in ear thickness (mm) ± S.E.M (n = 12) 2. Positive control

As shown in Table 3, when using the dew extract prepared according to the present invention, the arachidonic acid-induced stretching inhibitory effect is excellent, and particularly, when the concentration of the extract is 200 mg / kg, the arachidonic acid-induced stretching inhibition effect is found to be the most preferable. Can be.

Experimental Example 4: Acute Arthritis Treatment Effect Test

The acute arthritis treatment effect assay was performed on the dew extract prepared in Example 1, and the results are shown in Table 4 below.

Experimental Method 4

200 mg / Kg of S company J, pre-prepared in male SD rats (about 200 g), 1 ml oral administration of 100, 200, 400 mg / Kg of dew extract, 100 μl of 1% carrageenan saline solution and left plantar part Injection induced acute foot edema. 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% Whipped (100 mg / kg-succinic acid content 0.5% by weight) 66% Whipped (200 mg / kg-succinic acid content 1.0 wt%) 55% Whipped (400 mg / kg-succinic acid content 2.0 wt%) 57% Whipped (50 mg / kg-succinic acid content 0.25% by weight) 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 4, when the dew extract prepared according to the present invention is used, the acute arthritis treatment effect is excellent, and it may be seen that the acute arthritis treatment effect is particularly preferable when the dew extract 200 mg / kg concentration.

Experimental Example 5: Effect on NO and Prostaglandin E2 Production

The acute arthritis treatment effect assay was performed on the extract of dew drops prepared in Example 1 above.

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 hyssop extract on the production of prostaglandin E2 (PGE2). For measurement of NO, take 50 µl of the culture solution and mix 25 µl of Greases solution and measure absorbance at 540 nm. PGE2 was measured using an ELISA kit (R & D systems, USA).

After treatment with LPS (1 ㎍ / ml) and hyssop extract (25 ㎍ / ml) in raw 264.7 cell line, a mouse-derived macrophage, the culture medium was collected and quantified NO, an inflammation-inducing molecule in the body, by 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 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 level of the inflammation-related genes with respect to the dew extract prepared in Example 1 was compared to COX2 (Cyclooxygenase), iNOS (nitrogen Oxide Synthetase), mRNA expression changes in real time-PCR (Rotar gene 3000, Corbett) and Western blow Searching was done using Western blotting.

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

To investigate the anti-inflammatory effects of hyssop extract and its relationship with inflammation-associated molecules, we analyzed the effects of hyssop extract on COX2 (nitrogen oxide synthetase) genes and protein expression in real-time (Rotar gene 3000, Corbett). And western blotting.

Experimental Method 6-2] Measurement of mRNA Expression Change

LPS (lipopolysaccharide) (1 μg / ml) and bovine herbicides were treated with raw Qigen RNasy Kit from raw 264.7 cells cultured for 8 hours to synthesize cDNA, and then 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 treatment of LPS and E. coli herbal composition, proteins isolated from the cells were electrophoresed on 10% SDS polyacrylamide gel and the proteins on the gel were transferred to PVDF membrane under appropriate voltage. The membrane was blocked with tris buffer solution (25 mM Tris, pH 7.6, 192 mM NaCl) containing 10% skim milk and 0.5% BSA at room temperature for 2 hours. 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. The mRNA expression level of COX-2 increased about 66-fold after 8 hours of LPS treatment.

In addition, the mRNA expression of iNOS increased about 12-fold after 8 hours of LPS treatment, and the treatment with dew 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 hyssop extract, blood was drawn from the rabbit's abdominal artery using a syringe pre-filled with sodium citrate solution, centrifuged at 200 g for 10 minutes, and then platelet-rich plasma (PRP), the supernatant. Re-centrifuge some PRP for 10 minutes at 2.000 g to obtain Platelet Poor Plasma (PPP), a supernatant excluding precipitation, and the PRP to be used for platelet aggregation experiments using the 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 antiplatelet aggregation inhibitory effect was about 30% in the experimental group treated with 300 ㎍ / ml concentration of the herbal medicine composition.

Preparation Example 1 Preparation of Tablet

Using the herbal extract of the present invention, the tablet for oral administration was prepared by using the wet granulation method and the dry granulation method with the following composition.

[Furtherance]

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

Preparation Example 2 Preparation of Ointment

An ointment was prepared using the following extract of dew drops of the present invention with the following composition.

[Furtherance]

5 g of dew extract, 20 g of cetyl palmitate, 40 g of cetanol, 40 g of stearyl alcohol, 80 g of myristan isopropyl, 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

Injectables were prepared with the following composition using the extract of Emulsion of the present invention.

[Furtherance]

Emulsion extract 100mg, mannitol 180mg, monohydrogen phosphate 25mg, purified water for injection 2974 mg

Preparation Example 4 Preparation of Transdermal Agent

The percutaneous agent was prepared by the following composition using the dew extract of the present invention.

[Furtherance]

0.4 g of dew 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.

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 It is possible to achieve the standardization of the dew extract according to the control of the specific component and its content, such as acute edema suppression.

In addition, the extract of dew drops according to the present invention is prepared by extracting from the dew can be selected as succinic acid (succinic acid) as an indicator material can be prepared for the treatment of arthritis therapeutic activity by controlling the content of the indicator material.

Claims (3)

A pharmaceutical agent for the treatment and prevention of inflammatory diseases, characterized in that the extract is Amaranthaceae containing 0.5 to 5.0% by weight of succinic acid. The method of claim 1, wherein the dew extract is 1) extracting, cooling and filtering with 5-8 times the weight of wort protozoa by water, an alcohol or an aqueous alcohol solution; 2) separating the filtrate obtained above into an aqueous alcohol solution and concentrating under reduced pressure at 60 to 80 ° C., and 3) Concentrating the obtained filtrate, comprising the step of concentration under reduced pressure, vacuum drying, grinding and sterilizing the filtrate centrifuged at 500 ~ 1000 rpm after adding alcohol, A succinic acid (0.5% to 5.0% by weight of the drug) is characterized in that the powder. The agent according to claim 1, wherein the inflammatory disease is arthritis and edema.

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