KR20030086819A - Composition comprising sauchinone for treatment and prophylaxis of inflammatory diseases - Google Patents

Abstract

PURPOSE: Provided is a composition comprising, as an active ingredient, sauchinone for treatment and prophylaxis of inflammatory diseases. It has anti-inflammatory effect by inhibiting inducible expression of cyclooxygenase-2(COX-2). CONSTITUTION: A composition for treatment and prophylaxis of inflammatory diseases is characterized by comprising, as an active ingredient, sauchinone represented by the formula(I). The inflammatory diseases include articular rheumatism or asthma. The sauchinone is isolated and purified from the Saururs chinesis(Lour.) extract.

Description

Composition comprising sauchinone for treatment and prophylaxis of inflammatory diseases

The present invention has an anti-inflammatory effect through the inhibitory action of cyclooxygenase-2 (COX-2) -induced expression comprising a saucchinone compound of formula (I) as an active ingredient, A composition for the treatment and prevention of inflammatory diseases is provided:

(I)

The causes of chronic inflammatory diseases (eg, rheumatoid arthritis, asthma) arise from immune abnormalities in the body. For example, when the body's immune system removes them from the invasion of bacteria or viruses, it attacks the joints or parts of the body, causing symptoms of rheumatoid arthritis. About 1% of our population is serious enough to have rheumatoid arthritis, and further complications. Bronchial asthma is a disease that is accompanied by coughing, shortness of breath, tightness of the chest, and bronchial spasms. It is a common disease that accounts for 7-10% of the population. If the symptoms are severe, it is a condition that replaces all of the treatment with treatment such as bronchodilators temporarily. In recent years, the number of patients is increasing due to environmental pollution.

So far, nonsteroidal analgesic anti-inflammatory drugs and corticosteroids have been used for pain relief symptoms of chronic inflammatory diseases, but when taken chronically, they cause serious side effects such as gastrointestinal disorders and gastric ulcers. Recently, only cerebrex (Pfizer Pharmaceuticals, USA), which has a COX-2 inhibitory action, is used as a therapeutic agent for arthritis for the purpose of reducing such side effects. Therefore, the development of new medicines that can treat inflammatory diseases is urgent and urgent.

Therefore, the present inventors have been using a new candidate substance that can be developed as an inflammatory disease treatment for a long time to suppress inflammation and pain in the form of Chinese medicine or folk medicine, or isolated from natural products that have been described as having anti-inflammatory effects in medicine. Was intended. That is, in the present invention, the result of searching for the effect of inducing inflammation and removing the inflammation by using endotoxin material shows that the methanol extract of the ground part of 300 seconds shows a significant anti-inflammatory activity, from which the effect on the inflammation of macrophages The components present were followed and developed as new therapeutic agents for inflammatory diseases.

Three hundred herb is a perennial herb belonging to the family of three hundred (Saururaceae), and the whole grass or root or leaf is wind poison, diuresis, water species, gonorrhea, hepatitis, pneumonia, venom (便 毒), high blood pressure (高血壓) has been used in the private sector for the treatment. Three hundred seconds leaves, flowers and roots are white, and three leaves under the inflorescence (花序) of the upper part is called three hundred seconds (三 白草) because it turns white. Three hundred and four to five species of plants are distributed worldwide in over three hundred, and two out of two hundred species of three hundred vines belonging to three hundred vines in Korea, and two buckwheats of Houttuynia cordata Thunb . do. There are two species of genus Tricotaceae worldwide, Saururus chinensis in Asia and one species in North America ( Saururus cernuus , Lizard tail).

Three hundred seconds are stored in the ancient Chinese medicine book, Ming-Seok, which is collected in July and September, and dried in sunlight. Beauty is high, sex is cold, and enters the liver, lungs, and nerves. It is known to have the effects of moist heat, clearing, disinfection, and detoxification. Edema, each, jaundice, gonorrhea, and lobster It was used to treat back and back). In addition, the three hundred seconds root is harvested in Sinsuboncho, and in July-September, digging out the underground view to remove Ito, soaking water in boiling water, and drying it in the sun. Beauty is sensuous and sex is mild. Efficacy in mucus, dehumidification, clear fever and detoxification. It was used to treat individual, gonorrhea, lobulation, and improvement. .

Studies on the chemical composition of three hundred seconds have been conducted mainly on the flavonoids, alkaloids, amino acids, fatty acids, quinones and essential oils in the ground. It became. The essential oil component of the sentinel is methyl -n- nonyl-ketone (methyl- n -nonyl-ketone) was reported as the main component in addition to the alpha-pinene (a-piene), Kampen (camphene), Li nalrul (linalool), sapeu Safrol, beta-caryophyllene, humulene and the like have been reported. Flavonoids, one of the main components of the terrestrial sect, are reported to include hyperin, quercetin, isoquarcitrin, and quercitrin, and emodin and fish Quinone-based constituents such as physcion and saponin, lignans of carpanone-based, have also been reported. In addition, saurolactam, an aristolactam family of alkaloids, was isolated as a cytotoxic component of three hundred seconds, and the same series of alkaloids, aristotolactam B II, were also reported. It also contains fatty acids of cupric acid, linoleic acid and lauric acid, and alanine, valine, glutamic acid, aspartic acid and leucine. The presence of amino acids such as leucine, isoleucine and proline was also confirmed.

Until now, studies on the components of trichophytium showed that, among the components of trichophytium, sautinone showed hepatoprotective activity against carbon tetrachloride-induced hepatotoxicity, which was depleted of glutathione and antioxidant enzymes. (Sung, SH, Lee, EJ, Cho, JH, Kim, HS and Kim, YC, Biol. Pharm. Bull . 23, 666-668 (2000)). However, there are no reports of anti-inflammatory activity of triticale extract or its components.

The present inventors intensively studied the study of three hundred seconds, the extract of the three hundred seconds and the Sauchinone isolated from the anti-inflammatory effect through the inhibitory action of cyclooxygenase-2 (COX-2) -induced expression It confirmed to have for the first time and came to complete this invention.

Accordingly, it is an object of the present invention to provide a composition having a therapeutic and prophylactic effect of an inflammatory disease containing a sacchinone compound isolated from three hundred seconds as an active ingredient.

1 is a graph showing the effect of sacchinone on COX-2 expression inhibition,

2 is a graph showing the effect of Succinone on NF-kB activity induced by LPS,

3 is a graph showing the effect of Succinone on C / EBP activity induced by LPS.

4 is a graph showing the cell viability of sacchinone.

The present invention provides a composition for the treatment and prophylaxis of inflammatory diseases comprising sacchinone of formula (I) as an active ingredient:

(I)

The present invention also provides a composition for the treatment and prevention of inflammatory diseases, characterized in that the inflammatory disease is rheumatoid arthritis or asthma.

The present invention also extracts three hundred seconds from lower alkanols such as methanol, ethanol or propanol, chlorinated hydrocarbons such as methylene chloride or chloroform, hydrocarbons such as hexane or heptane, aromatic hydrocarbons such as benzene, toluene, or mixed solvents thereof. In addition, the present invention provides a composition for the treatment and prevention of inflammatory diseases comprising a three hundred seconds extract prepared by column chromatography as an active ingredient.

Hereinafter, the configuration and effect of the present invention will be described in detail.

It is a first aspect of the present invention to provide a composition for the treatment and prevention of inflammatory diseases, which comprises saucionone of formula (I) as an active ingredient:

(I)

Saucinone of the formula (I) of the present invention is isolated and purified in three hundred seconds.

Specifically, the present invention will be described below with an example of the extraction process of three hundred seconds.

Three hundred seconds is extracted by boiling water with a mixed solvent of a lower alkanol such as methanol, ethanol or propanol and a chlorinated hydrocarbon such as methylene chloride or chloroform, preferably a mixed solvent of chloroform and methanol (1: 1), and concentrated under reduced pressure to extract the total extract. Get Houttuynia cordata extract fraction thus obtained to the next, the chlorinated hydrocarbon is suspended in distilled water, and was again suspended in methanol, the resulting fractions, chlorinated hydrocarbons, and then, n - and fractions with hexane, methanol fraction and n - to obtain a hexane fraction. The obtained n -hexane fraction is subjected to silica gel column chromatography with a mixed solvent of n -hexane: ethyl acetate to obtain the sautinone of the present invention.

Succinone of the present invention may be prepared by methods known in the pharmaceutical art, in combination with conventional pharmaceutically acceptable carriers, excipients. Pharmaceutically conventionally acceptable pharmaceutical preparations can be formulated, for example, injections, solutions, syrups, tablets, capsules and the like.

The sacchinone of the present invention can also be converted to pharmaceutically acceptable inorganic or organic salts by conventional pharmaceutical methods. They can also be administered orally or parenterally.

Succinone of the present invention is appropriately selected according to the absorption of the active ingredient in the body, the rate of excretion, the age and weight of the patient, sex and condition, the severity of the disease to be treated, etc., generally 10 mg to 5,000 mg per day in adults May be administered 1 to 3 times, and the amount may be increased or decreased depending on the weight, sex, age, and severity of the patient.

The second aspect of the present invention is based on three hundred seconds in a lower alkanol such as methanol, ethanol or propanol, chlorinated hydrocarbons such as methylene chloride or chloroform, hydrocarbons such as hexane or heptane, aromatic hydrocarbons such as benzene, toluene, or a mixed solvent thereof. It is to provide a composition for the treatment and prophylaxis of inflammatory diseases extracted or extracted from the three hundred seconds extract prepared by column chromatography.

The triticale extract of the present invention is also commonly known by the methods known in the pharmaceutical field with a pharmaceutically acceptable carrier, excipient. Pharmaceutically conventionally acceptable pharmaceutical preparations can be formulated, for example, injections, solutions, syrups, tablets, capsules and the like. They may be administered orally or parenterally, and in general, adults may be administered 10 mg to 10,000 mg once or three times daily, and may be increased or decreased depending on the patient's weight, sex, age and degree of disease. have.

Hereinafter, the present invention will be described in more detail with reference to the following examples. These examples are for illustrative purposes of the present invention and are not intended to limit the protection scope of the present invention.

Example 1. Extraction and Fraction of Three hundred Seconds

The dried three hundred seconds 10kg was extracted three times for two hours with a mixed solvent of chloroform and methanol (1: 1), and concentrated under reduced pressure to give a total extract of 1.8kg. The triticale extract (1.8 kg) was suspended in distilled water, fractionated with chlorinated hydrocarbon and concentrated under reduced pressure to obtain a chlorinated hydrocarbon fraction and a water fraction. It was re-suspended in 90% methanol, a chlorinated hydrocarbon fractions and then concentrated, n - the fraction with hexane to 90% methanol fraction (120g) and n - to give a hexane fraction (60g).

Example 2 Isolation and Identification of Succinone

The n -hexane fraction (60 g) was separated into 20 small fractions (Fr. 1-20) by silica gel column chromatography with a mixed solvent of n -hexane: ethyl acetate (100: 1 to 0: 1). Six fractions (2 g) were separated by Sephadex LH-20 column chromatography using a mixed solvent of n-hexane: chlorinated hydrocarbon: methanol (5: 1: 1). )

Determination of the Structure of Sauuccinon

C ₂₀ H ₂₀ O ₆

[α] _D ; + 95.0 ° (c, 0.63; CHCl ₃ ),

R _f ; 0.35 (n-hexane: EtOAc = 5: 1),

UV (CHCl ₃ ) λ _max (log ε); 299.5 (2.55), 254.2 (2.74) nm,

IR (neat) ν _max ; 2916, 1676, 1664, 1418, 1433, 1321, 1241, 1184, 1155, 979, 926, 892, 756 cm ^-1 ,

EIMS ( m / z ) (rel. Int.); 356 [M ⁺ ] (100), 270 (13), 257 (18), 205 (26), 175 (57), 151 (39), 138 (23),

HR EIMS; m / z 356.1262 (calcd for C ₂₀ H ₂₀ O ₆ m / z 356.1260),

¹ H NMR (300 MHz, CDCl ₃ ) δ 6.82 (1 H, s, H-6), 6.38 (1 H, s, H-3), 5.90 (1 H, d, J = 1.40 Hz, aromatic -OC H ₂ O-), 5.87 (1 H, d, J = 1.40 Hz, aromatic -OC H ₂ O-), 5.65 (1 H, s, aliphatic -OC H ₂ O-), 5.60 (1 H, s, aliphatic -OC H ₂ O-), 5.57 (1 H, s, H-3 '), 3.03 (1 H, d, J = 5.40 Hz, H-7), 2.52 (1 H, td, J = 11.93, 3.47 Hz, H-1 '), 2.48 (1 H, d, J = 5.40 Hz, H-6'), 2.44 (1 H, m, H-8), 1.92 (1 H, m, 7'-H _ax ), 1.88 (1 H, m, H-8 '), 1.64 (1 H, m, 7'-H _eq ), 1.22 (3 H, d, J = 7.26 Hz, H-9), 0.71 (3 H, d, J = 7.39 Hz, H-9 '),

¹³ C NMR (100 MHz, CDCl ₃ ) δ 199.69 (C-2 '), 168.66 (C-4'), 146.72 (C-5), 145.70 (C-2), 143.26 (C-4), 115.73 ( C-1), 105.54 (C-6), 101.34 (C-3 '), 101.24 (aromatic -O C H ₂ O-), 100.42 (C-5'), 99.50 (C-3), 98.19 (aliphatic -O C H ₂ O-), 37.60 (C-6 '), 37.55 (C-1'), 35.06 (C-7), 34.82 (C-8), 33.45 (C-8 '), 25.27 (C -7 '), 21.29 (C-9), 20.92 (C-9')

The results of spectroscopy analysis were compared with the literatures (Sang Hyun Sung and Young Choong Kim, J. Nat. Prod. 63 (7), 1019-1021 (2000)) and identified as Sauchinone.

Experimental Example: Screening for Inflammatory Defense Activity of Macrophages

In the process of creating a substance with macrophage inflammatory defense activity from natural products, the first requirement is the establishment of a suitable screening method to search for substances having such activity. Macrophages of rats known to be physiological and biochemically similar to human macrophages have been widely used as search systems in order to find substances having inflammatory defense activity from natural products. In order to detect macrophage protection activity in natural products, the natural macrophages are treated with natural products, and after a certain time, endotoxins are treated to measure the degree of protective effect when artificially inflamed.

In the present invention, lipopolysaccharide (LPS) was used to induce inflammation in macrophages. LPS is a major component of the Gram-negative bacterial cell wall, and it binds to the plasma LPS binding protein (LPB) in the cytoplasm and moves to the phospholipid of the cell membrane to bind to CD14 present on the surface of the macrophage, or 95kDa in which the LPS itself is present on the cell membrane , 80kDa protein and the like, and inflammatory response (Hewett, JA and Roth, RA: Hepatic and extrahepatic pathobiology of bacteriallipopolysaccharides.Pharmacol . Rev. 45, 382-411 (1993)). The binding of LPS to the receptor stimulates intracellular G1 protein and results in tumor nacrosis factor (TNF-a), interleukin- and TNF-a signaling from cells through signaling systems of mitogen activated protein kinase (MAPK). Various inflammatory mediators such as cytokines such as 1 (IL-1), IL-6, prostanoids, leukotriens and nitro oxides (NO) do.

In this experimental example, to determine the anti-inflammatory effect of inhibition of cyclooxygenase-2 (COX-2) in macrophages, the protective effect against LPS was measured at the mRNA and protein levels of cyclooxygenase-2. , COX-2 transcription factor activity was measured. In addition, cell viability was measured by MTT [3- (4,5-dimethylthiazole-2-yl) -2,5-diphenyl-tetrazolium bromide] assay to observe the toxicity of sacchinone. The data shown in this experiment was analyzed using a pharmacologic calculation program. Significance between the various treatment groups was tested by one way analysis of variance (ANOVA) followed by Newmann-Kelus test (** p <0.01).

(1) RAW264.7 cell culture, mouse macrophage line

RAW264.7 cells are mouse macrophage lines (Korea Cell Line Bank, Seoul, Korea), Dulbeco's Modified Eagles medium containing 10% fetal bovine serum, 100 U / ml penicillin and 100 mg / ml streptomycin Incubated at. 1 to 2 × 10 ⁶ / ml of RAW264.7 cells were pre-incubated for 24 hours in a 37 ° C. chamber containing 5% CO ₂ and 95% air. All experiments maintained 80-90% confluency and did not exceed 20 passages.

(2) Determination of the Effect of Succinone on Inhibition of COX-2 Expression

2-1) Northern blot analysis

The RAW264.7 cells incubated in (1) were dissolved in dimethylsulfoxide in dimethylsulfoxide and treated at concentrations of 1, 3, 10 and 30 μM. After 1 hour, LPS (Escherichia coli 0111) was applied to the RAW264.7 cells. : B4; Sigma, St. Louis, MO) was treated with 1 μg / ml. Total RNA from RAW264.7 cells thus treated was isolated by the method of Chomczynski and Sacchi. The total RNA isolated was dissolved in sample dilution buffer (50% formamide, 2.2M formaldehyde in 1 × MOPS buffer was adjusted). RNA was denatured at 65 ° C. for 15 minutes. Electrophoresis was performed on 1% agarose gel. After electrophoresis, RNA present in the gel was transferred to nitrocellulose paper, and the transferred nitrocellulose paper was dried in a vacuum oven at 80 ° C.

Denatured COX-2 cDNA was randomly primed at ³² P and used as a probe. Hybridization was followed by hybridization of nitrocellulose paper at 42 ° C. for 2 hours in a hybridization buffer containing 50% deionized formamide, 5 × Denhardt's solution, 0.1% SDS, ssDNA, 5 × SSPE. Labeled probes were added and incubated at 42 ° C. for 18 hours, then washed twice at 42 ° C. for 15 minutes each with 2 × SSC / 0.1% S DS and 0.1 × SSC / 0.1% SDS and 0.1 × SSC / 0.1% SDS was washed at 55 ° C. for 1 hour and the results are shown in FIG. 1A.

2-2) Immunochemical Analysis

Cells were scraped with a scraper in a Raw264.7 cell line treated with Succinone and LPS as described in 2-1), and then used as a lysis buffer [20 mM Chloride Tris (pH 7.5), 1% Triton X-100, 137 mM sodium chloride, 10% glycerol, 20 mM EDTA, 1 mM sodium orthovanadate, 25 mM beta glycerophosphate, 2 mM sodium pyrophosphate, 1 mM phenylmethylsulfonylfluoride 100 μl of 1 μg / ml leupeptin] was placed on ice for 1 hour and centrifuged at 10,000 g for 10 minutes to obtain supernatant to separate lysate. And then transferred to nitrocellulose to bind an anti-mouse COX-2 antibody (1: 1000 in 1 × PBS), and the secondary antibody was an alkaline-phosphatase-conjugated anti-goat. ) Antibody (1: 1000 in 1 * PBS) Use was incubated at room temperature. 5-bromo-4-chloro-3-indolyl was coloring to a phosphazene-Tate / 4-nitro blue tetrazolium chloride, and the results were shown in Figure 1b.

As shown in Figures 1a and b, after 1 hour pre-treatment of sacchinone, LPS was added and the expression changes of COX-2 mRNA and protein were observed. As a result, COX- against LPS at concentrations of 1-30 μM of sacchinone was observed. 2 mRNA and protein expression was suppressed by 90%. That is, sacchinone selectively inhibited COX-2 involved in the inflammatory response.

(3) Gel retardation assay

Nuclear factor-kappa B (NF-kB) is present in the promoter region of the gene and is activated by LPS to increase COX-2 gene expression. In order to determine the effect of Succinone on NF- k B activity induced by LPS, gel shift analysis was performed to determine the effect of NF- k B DNA binding at 3 μM concentration of Succinone. In addition, C / EBP (CCAAT / enhancer binding protein) is present in the promoter region of the COX-2 gene and is activated by LPS to increase gene expression. To determine the effect of Saucinone on C / EBP activity induced by LPS, nucleoprotein was isolated from RAW264.7 cells to determine the effect of C / EBP DNA binding at 3 μM concentration of Sauchinone. gel shift) analysis.

The specific experimental method is as follows.

Gel shift analysis was performed by end-labeling consensus sequences of NF-kB and C / EBP. Lysis buffer [10mM HEPES (pH 7.5), 10mM potassium chloride, 0.1mM EDTA, 1mM dithiothreitol in Raw264.7 cells treated with Succinone (3 μM) and LPS as in 2-1) ), 0.5mM phenylmethylsulfonyl fluoride)] and left on ice for 10 minutes, centrifuged at 7,200g for 6 minutes to remove the supernatant and extraction buffer [20mM HEPES (pH7.9), 400mM sodium chloride , 1mM EDTA, 1mM dithiothreitol, 1mM phenylmethylsulfonyl fluoride] and left on ice for 30 minutes, centrifuged at 15,000g for 10 minutes to take the supernatant to separate the nuclear protein. Isolated nucleoprotein and 5 × binding buffer (20% glycerol, 250 mM EDTA, 2.5 mM dithiothreitol (DTT), 0.25 mg / ml poly dI-dc, 50 mM Tris-Cl (pH) 7.5)) for 10 minutes after pre-reaction at room temperature, by the addition of a labeled probe with NF- k B, C / EBP consensus sequence with a radioactive isotope and reacted for 20 minutes at room temperature. The reaction mixture was electrophoresed on a 4% polyacrylamide gel in 0.5% TBE buffer to fix the gel, followed by autoradiography. The effect of Succinone on NF-kB activity induced by LPS is shown in FIG. 2 and the effect of Succinone on C / EBP activity induced by LPS is shown in FIG. 3, respectively.

As shown in Figure 2, the activity of NF-kB induced by LPS was inhibited by sacchinone. In addition, LPS binds p50, which is the constituent protein of NF-kB, and p65 binds to p65, and Sauchinone inhibits the migration of p65 protein into the nucleus, thereby inhibiting NF-kB activity. In addition, as shown in FIG. 3, C / EBP activity induced by LPS was inhibited by sacchinone.

As described above, it can be seen that the saucchinone compound of the present invention has an anti-inflammatory effect through the inhibitory action of the induced expression of cyclooxygenase-2 (COX-2).

(4) measurement of viability of cells (MTT assay)

Cell viability was measured by MTT assay to observe the toxicity of sacchinone. The RAW264.7 cells cultured in (1) were placed in 96 wells so as to be 1 × 10 ⁵ / ml cell / well, and treated with sacchinone at concentrations of 1, 3, 10, 30, and 100 μM (control). Uchinone was treated at concentrations of 1, 3, 10, 30 and 100 μM and 1 hour later LPS (1 μg / ml) (LPS + Sauchinone) at 37 ° C. containing 5% CO ₂ and 95% air. The reaction was carried out in the chamber for 24 hours. RAW264 with Succinone (1, 3, 10, 30 and 100 μM) and LPS. For 7 cells, formazone (formazon) generated by adding MTT was read at 540 nm with a Titertek Multiskan Automatic ELISA reader to confirm cell viability, and the results are shown in FIG. 4. The group treated with Succinone alone was used as a control.

As shown in FIG. 4, the cell viability was not affected until 24 hours in the group treated with sacchinone at concentrations of 1, 3, 10, 30 and 100 μM and treated with LPS.

Next, the present invention will be described in more detail as formulation examples.

Formulation Example 1

Succinone 100mg

Appropriate sterile distilled water for injection

pH adjuster

Succinone is dissolved in distilled water for injection, adjusted to pH 7.6 with a pH adjusting agent, and then the total amount is 2 ml, and then filled into 2 ml ampoules and sterilized to prepare an injection.

Formulation Example 2

Succinone 10mg

Lactose 100mg

Starch 50mg

Magnesium stearate proper amount

The above components are mixed and tableted according to a conventional method for producing tablets to produce tablets.

Formulation Example 3

Sauchinone 5mg

Lactose 100mg

Starch 93mg

Tackle 2mg

Magnesium stearate proper amount

The capsules are prepared by mixing the above components and filling gelatin capsules according to a conventional method for preparing capsules.

Formulation Example 4

Succinone 100mg

20 g of sugar

20 g of isomerized sugar

Lemon flavor

Add 100 ml of purified water

The above components are mixed according to a conventional method for preparing a liquid, and filled into 100 ml brown bottles and sterilized to prepare a liquid.

As can be seen from the above, the sacchinone and trichococcum extract of the structural formula (I) of the present invention exhibit a cyclooxygenase-2 (COX-2) inhibitory action, thus preventing and treating new inflammatory diseases, particularly rheumatoid. It can be used as a prophylactic and therapeutic agent for chronic inflammatory diseases such as arthritis and asthma.

Claims (3)

A composition for the treatment and prevention of inflammatory diseases comprising a sautinone of formula (I) as an active ingredient: (I) According to claim 1, wherein the inflammatory disease is a composition for the treatment and prevention of inflammatory diseases, characterized in that rheumatoid arthritis or asthma. Three hundred seconds is extracted from lower alkanols such as methanol, ethanol or propanol, chlorinated hydrocarbons such as methylene chloride or chloroform, hydrocarbons such as hexane or heptane, aromatic hydrocarbons such as benzene, toluene, or a mixed solvent thereof, or the like Composition for the treatment and prophylaxis of inflammatory diseases comprising the extract of the three hundred seconds by chromatography.