KR20070117889A - A pharmaceutical composition comprising the extract of artemisa spp for treating or preventing inflammatory bowel disease - Google Patents

Abstract

A pharmaceutical composition comprising the extract of Artemisa spp. is provided to reduce expressions of iNOS(inducible nitric oxide synthase), TNF-alpha(tumor necrosis factor-alpha), COX-2(cyclooxygenase-2), IFN-gamma(interferon-gamma), STAT1(signal transducers and activation of transcription 1) and p-STAT1, so that it is useful for treating or preventing inflammatory bowel disease. A pharmaceutical composition for treating or preventing inflammatory bowel disease comprises the extract of Artemisa spp. having anti-inflammatory effects which is prepared by drying and pulverizing Artemisa spp., dissolving the pulverized Artemisa spp. in 1% phosphate buffered saline, centrifuging the solution, and filtering the centrifuged solution, and is formulated as solution, syrup, tablet or capsule. The daily dosage of the Artemisa spp. extract is 100-500 mg/kg.

Description

Pharmaceutical composition comprising the extract of Artemisa spp for treating or preventing inflammatory bowel disease

1 shows the effect on Raw 264.7 cell viability when LPS-induced inflammation in Raw 264.7 cells and concentration-dependent treatment of the extract of strengthening wormwood of the present invention.

Figure 2 shows the effect on nitrite (NO) of Raw 264.7 cells of the fortified wormwood extract of the present invention.

Figure 3 shows the effect on iNOS, COX-2 protein expression when treated with concentration-dependent treatment of the strengthening wormwood extract of the present invention.

Figure 4 shows the effect of the enhanced wormwood extract of the present invention on iNOS, COX-2 mRNA expression.

Figure 5 shows the effect of the enhanced wormwood extract of the present invention on the expression of TNF-α.

Figure 6 shows the effect of the enhanced wormwood extract of the present invention on the expression of STAT1, p-STAT1.

Figure 7 shows the effect of the enhanced wormwood extract of the present invention on IFN-γ expression.

The present invention relates to a pharmaceutical composition for the prevention and treatment of inflammatory bowel disease, including fortified wormwood extract, the fortified wormwood extract of the present invention is excellent in the protective effect of the body in vivo as a prophylaxis and treatment of inflammatory bowel disease Can be used.

Ganghwa wormwood ( Artemisa spp ) is called lion foot wormwood because the appearance of wormwood leaves is divided into lion foot shape and the node grows with white hairs on the back. This strengthened medicinal plant is rich in organic matter, and if it is well drained, it grows in the foot of 100 m above sea level on sunny beaches, and the leaf and rhizome contain the medicinal ingredients with different effects, preventing and treating various diseases. It is widely used as. It grows up to about 70 cm in height, gives off a distinctive scent, and contains vitamins A, B, C, carotene, cineol, amylase, etc., and its effects include hemostasis, antibacterial, gastrointestinal diseases, respiratory diseases, etc. It is known that there is.

Inflammatory bowel disease (IBD) is a disease that causes chronic inflammation in the gastrointestinal tract, and is commonly classified into two types: ulcerative colitis (UC) and Crohn's disease (CD). Important immunological factors in the cause and treatment of the inflammatory bowel disease include innate immunity, production of cytokines, activation of CD4, and the like. Evidence for the role of cytokines in the invention and research of inflammatory bowel disease is clear.

Cytokines are important factors influencing the regulation of inflammation and immune responses. Recently, studies on the production of pro-inflammatory cytokine have been made. In addition, tumor necrosis factor (TNF-α), Interluekin (IL) -1, IL-6, and IL-8 production at the site of inflammation were significantly increased in ulcerative colitis and Crohn's patients. It became. Studies on the regulation of cytokine production in patients show that the pathophysiology is very important in these diseases (Sartor RB, Cher, D. J et al ., 1987).

IFN-γ, IL-2, IL-4 and IL-10 are important factors in the initiation, control and function of inflammation in ulcerative colitis and Crohn's disease (Lin CC et al ., 1996; Kojouharoff et al ., 1997; Obermeier F et al ., 1999; Neurath MF et al ., 1995; Schreiber S et al ., 1995; Seder RA et al ., 1993). As such, immunological factors are important in the treatment as well as the cause of the development of inflammatory bowel disease.

The establishment of animal models of inflammatory bowel disease is essential for clinical trials. In order to establish an inflammatory bowel disease animal model, exogenous agents such as dextran sodium sulfate (DSS) are used, and IL-2, IL-10, and T cell receptor deficient mice. Although there are many ways of genetically engineered to make T cell receptor-deficient mice, it has been reported that the animal model that caused ulcerative colitis with 5% dextran sodium sulfate (DSS) is the best model.

In addition, animal models of colitis induced by DSS showed symptoms similar to those of ulcerative colitis in humans.

On the other hand, macrophages play several roles in the immune response, which play an important role as antigen presenting cells (APCs) when exposing antigens to lymphocytes in the process of generating specific immune responses. By secreting a variety of substances, it plays a role in the immune response caused by lymphocytes.

Macrophages have the property of reacting when stimulated with stimulants such as lipopolysaccharides (LPS), which are of physiological and pharmacological interest. When macrophages react to these substances, they produce a variety of substances, including cytokines, ranging from 32 to 440 kDa, and secrete cancer cell lytic enzymes such as cytolytic proteolytic enzymes and tumor necrosis factor (TNF-α).

Accordingly, the present inventors have continued efforts to develop new prophylactic and therapeutic agents for inflammatory bowel disease. As a result, we have confirmed that fortified wormwood extract has an excellent protective effect on inflammatory bowel disease and cell inflammation, and successfully completed the present invention.

An object of the present invention is to provide a pharmaceutical composition for the prevention and treatment of inflammatory bowel disease and cell inflammation with an extract of the Artemisa spp having an anti-inflammatory effect and a method for producing the same and its preparation method as an active ingredient. .

In order to achieve the above object, the present invention provides fortified wormwood extract ( Artemisa spp ).

In addition, the present invention provides a use of the fortified wormwood extract as an anti-inflammatory agent.

The present invention also provides a use of the fortified wormwood extract as a therapeutic agent for inflammatory bowel disease and bacterial inflammation.

The present invention also provides a pharmaceutical composition for anti-inflammatory, containing the fortified wormwood extract as an active ingredient.

The present invention also provides a pharmaceutical composition for the prevention and treatment of inflammatory bowel disease and bacterial inflammation containing the fortified wormwood extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides an enhanced wormwood extract ( Artemisa spp ) having an anti-inflammatory effect.

In addition, the present invention provides a use of the fortified wormwood extract as an anti-inflammatory agent.

The present invention also provides a use of the fortified wormwood extract as a therapeutic agent for inflammatory bowel disease and bacterial inflammation.

The present invention also provides a pharmaceutical composition for anti-inflammatory, containing the fortified wormwood extract as an active ingredient.

The present invention also provides a pharmaceutical composition for the prevention and treatment of inflammatory bowel disease and bacterial inflammation containing the fortified wormwood extract as an active ingredient.

Fortified wormwood extract of the present invention has the effect of reducing iNOS, TNF-α, COX-2, IFN-γ, STAT1, p-STAT1 expression and inhibiting inflammation when macrophage cells such as Raw 264.7 cells are induced have. Therefore, the extract of the strengthening wormwood of the present invention is proved to be excellent in the anti-inflammatory effect.

The pharmaceutical compositions of the present invention may be used with pharmaceutically acceptable carriers or excipients according to their use and may be formulated into all other forms suitable for administration to a person, for example liquids, syrups, tablets and capsules. Can be. Carriers that may be contained in the pharmaceutical compositions of the present invention may include extenders, high fiber additives, encapsulating agents, lipids, and the like, examples of which are well known in the art.

In addition, the pharmaceutical composition of the present invention may be administered in varying dosages depending on the extent of disease progression, age, sex, physical condition, duration of administration, administration method, weight of the patient, diet, rate of discharge, and the like. Preferably, fortified wormwood extract contained in the pharmaceutical composition may be administered orally 1 to 3 times a day in the range of 100 to 500 mg.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited.

Example 1. Preparation of Fortified Wormwood Extract

In the present invention, in order to obtain Artemisa spp , after drying powdered wormwood in the shade and crushed with a crusher to powder, the powdered wormwood powder 10% in 1% Phosphate buffered saline (PBS) solution It was dissolved at a concentration of / ㎖, centrifuged at 10,000 × g and filtered to prepare a powdery wormwood extract.

Example 2 Preparation of Experimental Animals

Raw 264.7 cells were cultured using 10% Fetal bovine serum (FBS), 1% penicillin streptomycin DMEM medium (Dulbecco's modified eagle's medium).

Four-week-old Balb / c female rats were acclimated to a dark-light cycle for 12 hours and dextran sodium sulfate (DDS) dissolved in water at a concentration of 0.5% under a controlled condition of 25 W 2 ° C. was induced for 1 week. After induction, the fortified wormwood extract prepared in Example 1 was administered orally at a predetermined time every day.

The rats were anesthetized and then opened, and spleen and MLN were extracted and used for the experiment.

Experimental Example 1. Macrophage stimulation experiment by LPS

The present invention is to determine the effect on the cell proliferation after activating Raw 264.7 cells with fortified wormwood extract, in order to confirm the macrophage (macrophage) stimulation by lipopolysaccharide (LPS), two-day culture of Raw 264.7 cells One was used.

LPS was diluted using phosphate buffer solution (PBS; 0.45 μm filted) or serum free DMEM medium (add penicillin-streptomycin and L-glutamine).

24 wells / plate was washed twice with incomplete DMEM medium or incomplete RPMI 1640 medium, and then 1.5 ml or 1.0 ml of serum free DMEM medium (added penicillin-streptomycin and L-glutamine) was added and LPS was added for 12 hours. Or stimulated in a CO ₂ incubator for 24 hours.

Stimulated medium was transferred to 6 ml tubes, stored at -20 ° C until use, and used for experiments on Raw 264.7 cell viability, ELISA, immunoblotting, and macrophage conditioned cells remaining after media removal. ) Was immediately washed with cold HBSS, stored at -20 ° C and used for immunoblotting.

Experimental Example 2. Cell Survival Rate Experiment

In order to examine the effects of nitrate produced by cytokines on Raw 264.7 cells, MTT was performed. This method uses 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT) to promazan (fromazan) by succinate-dehydrogenase, an enzyme in the cell's mitochondria. It is an experiment that uses a decrease in the production of promazan when the cell stops growing or dies.

First, 1 × 10 ⁵ cells / ml of Raw 264.7 cells were dispensed in 1 ml portions of 24 wells / plates and incubated for 24 hours in a 37 ° C., 5% CO ₂ incubator.

The cultured cells were treated with 40, 60, 80, and 100 mg / ml of the fortified wormwood extract prepared in Example 1, and cultured for 48 hours, followed by 5 mg / ml MTT dissolved in PBS (Sigma Chemical, St. Louis, MO, USA) 50 ml of the solution was added to each well and incubated for 4 hours at 37 ℃, 5% CO ₂ incubator. After incubation, the culture solution was discarded, and 500 µl of 0.04 N HCl dissolved in isopropyl alcohol (IPA) was added to dissolve promazan, followed by 550 nm with a microplate reader (Model 550, Bio-Rad labolatories, Japan). Absorbance was measured at.

As a result of measuring the cell viability of the experimental group activated with fortified mugwort extract with 100 as the proliferation of cells cultured for 24 hours in an unstimulated medium, as shown in FIG. Activated experimental group increased cell viability.

In addition, as the concentration of the strengthening wormwood extract increased cell survival rate, especially when the concentration of the strengthening wormwood extract was confirmed that the maximum number of cells increased.

Experimental Example 3. Measurement of nitrite

The Griess reaction was used to measure the nitrites generated in Raw 264.7 cells by cytokines. This method uses a color reaction in which the diazo group of the Griess reagent turns pink when it meets nitrite.

⁶ ml of raw 264.7 cells / ml 6 ml / ml in 6 well plates were planted in DMEM medium containing no phenol red and incubated in a 37 ° C., 5% CO ₂ incubator for 24 hours. It was incubated with time zone by treatment with fortified wormwood extract prepared in Example 1 and LPS.

After incubation, 100 μl of the supernatant was measured in 96 wells / plate for nitrite measurement, and reacted for 10 minutes in a dark place at room temperature by adding an equal amount of Griess reagent (Sigma Chemical, St. Louis, MO, USA). Absorbance was measured at 550 nm with a microreader. The nitrite concentration was calculated by comparing with a standard straight line obtained using sodium nitrite (NaNO ₂ ), and the results are shown in FIG. 2.

Nitrogen monoxide (NO) is one of the free radicals and is a very unstable molecule. Nitrogen monoxide is converted to stable nitrogen oxides such as NO ₂ , N ₂ O ₃ , N ₂ O ₄ , nitrite (NO ^2- ) and nitrate (NO ^3- ) by oxygen or superoxide. . Nitrogen monoxide is produced by nitric oxide synthase (NOS) from L-arginine, which is composed of endothelial NOS and neuronal NOS (nNOS), which produce nitric oxide, which is required for homeostasis. Or iNOS induced by inflammatory tension or the like.

As a result of the concentration-dependent treatment of the production of nitrite by varying the concentration of the extract of Fortified Mugwort in the culture medium of Raw 264.7 cells, the production of Nitrite was increased in the LPS group, whereas the amount of Nitrite was added to the control group according to the present invention. (CON group) was confirmed to decrease (see Fig. 2).

That is, it was confirmed that when the concentration of the extract of strengthened wormwood extract is 100 ㎍ / ㎖ nitrite production was reduced nitrite production than LPS 1 ㎍ / ㎖ showed a strong immune response.

Experimental Example 4. Measurement of iNOS and COX-2 Protein Expression by Western Blotting Analysis

Experiments were performed when more than 80% of the cells were grown after dispensing 1 × 10 ⁶ cells / ml of raw 264.7 cells into 6 well plates.

Divided into control group, LPS group and LPS + fortified wormwood group (40, 60, 80, 100 ㎍ / ㎖), incubated for 24 hours, washed with PBS, and then dissolved in a buffer solution (cell), 50 mM HEPES (pH 7.0), 250 mM NaCl, 5 mM EDTA, 0.1% Nondiet P-40, 1 mM PMSF, 0.5 mM Dithiothreitol (DTT), 5 mM NaF, 0.5 mM Na Orthovavadate, Leupeptin and Aprotinin for 30 minutes at 4 ° C. Incubated.

Perform the Bio-Rad protein assay with the cell supernatant, load 8-12% polyacrylamide gel electrophoresis for 100 V for 1 hour, and then 100 V for 1 hour on the nitrocellulose membrane. Transfer was carried out for 30 minutes.

Blocking was performed for 1 hour with 5% skim milk powder, and primary antibodies of 1: 1000 TNF-α, COX-2, iNOS, p-STAT1, STAT1, and IFN-γ were overnight.

After washing 4 times with Tween 20 / Tris buffer (TTBS) for 5 minutes, the secondary antibody was attached for 2 hours, iNOS and COX-2 protein expression levels were confirmed using ECL kit, and the results are shown in FIG. 3. Indicated.

As shown in Figure 3, in the case of iNOS, the control group (CON group) did not appear iNOS protein expression level, it was apparent in the LPS group. In addition, it was confirmed that the protein expression amount is reduced also in the fortified wormwood group to which the fortified wormwood extract is added.

Similarly in the COX-2, the control group did not show the protein expression level, but the LPS group showed the expression level. In addition, even in the fortified wormwood group, the protein expression amount was not significantly reduced but was confirmed to decrease.

Experimental Example 5. Measurement of TNF-α, iNOS and COX-2 Gene Expression by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) Method

1 × 10 ² cells / ml Raw 264.7 cells were grown in 100 mm dishes with growth medium, incubated for 24 hours, treated with fortified wormwood extract, and the supernatant was discarded to separate RNA and 1 ml of TRIzol reagent (Invitrogen Life). Technologies, USA) was added to the reaction in three minutes at room temperature and collected in a 1.5 ml tube.

200 μl of chloroform was added thereto, shaken well for 15 seconds, and then stored in ice for 10 minutes. After centrifugation at 12,000 × g for 15 minutes at 4 ° C., the supernatant was transferred to a previously cooled 1.5 ml tube, and the same amount of cold IPA was added thereto and then incubated at −20 ° C.

The supernatant was removed by centrifugation at 12,000 × g for 20 minutes at 4 ° C., and 1 ml of 75% ethanol was added to the RNA pellet.

Again, the supernatant was removed by centrifugation at 12,000 × g for 5 minutes at 4 ° C., and the RNA pellet was dried for about 30 minutes on a clean bench, then treated with diethylpyrophosphate (DEPC). It was dissolved in 23 μl of distilled water.

Quantification of RNA was diluted 50-fold and absorbance was measured at 260 nm using a UV / Vis spectrophotometer (SmartSpec ™ 3000, Bio-Rad, USA).

1 μl of cytokine 20 pM / μl sense primer (Bioneer, Seoul, South Korea) and 20 pM / μl antisense primer (Bioneer, Seoul, South Korea) on RT / PCT premix (Bioneer, Seoul, Korea) 1 μl, 1 μg RT product, DEPC treated distilled water was added to the final volume of 50 μl, and then PCR was performed.

RT-PCR showed cDNA synthesis: 42 ° C., 60 minutes; predenaturation: 95 ° C., 5 minutes; denaturation: 95 ° C., 1 min; annealing: 55-65 ° C., 1 min; elgation: 72 ° C., 1 min; repeated 31 times, followed by 1% agarose gel with postelngation bromide (EtBr; Sigma Chemicals, St. Louis, MO, USA) (Cambrex Bio Science Rockland Inc., Rockland, ME, USA) was electrophoresed at 75V for 40 minutes and observed in UV.

At this time, the forward primer of TNF-α used is 5'-GCGACGTGGAAGTGGCAGAAG-3 ', the reverse primer is 5'-TCCATGCCGTTGGCCAGGAGG-3', the forward primer of iNOS is 5'-CCTTGTTCAGCTACGCCTTC-3 ', the reverse primer is 5'-CTGAGGGCTTGCTTGAGGTC -3 ', forward primer of COX-2 was 5'-TGCATGTGGCTGTGGATGTCAT-3', and reverse primer was 5'-CACTAAGACACCCGTCATCTCCA-3 '.

As shown in FIG. 4, the mRNA expression levels of iNOS and COX-2 were decreased by performing reverse transcriptase polymerase chain reaction (RT-PCR). .

In iNOS, the expression level of mRNA was found in the LPS group, and the expression level decreased as the concentration of the strengthened wormwood group increased. COX-2 also showed the same trend.

Therefore, when 100 μg / ml of the extract of Fortified Mugwort was added, the amount of mRNA expression decreased the most.

Example 6. TNF-α Assay Using ELISA

(1) Preparation of rabbit anti-mouse TNF-α antibody

Assay diluent bottles were mixed with a vortex, 3.5 ml were taken and placed in lyophilized rabbit anti-mouse antibody and mixed by rotatory.

(2) Preparation of TNF-α Standard Solution

(A) Sample preparation

Five kinds of TNF-α standard solutions were prepared, 840 μl was added to # 1, and 759 μl of assay dilution was added to # 2 to # 5, and the mixture was mixed with a rotator. At this time, the concentration of the TNF-α standard solution was 10,000 pg / ㎖, 160 μl was taken into standard tube # 1 containing the assay diluent. # 2 was prepared by taking 250 μl of # 1, and # 3 to # 4 were prepared in the same manner. Thus, the concentration of each standard tube was # 1 at 1, 600 pg / ml; # 2 is 400 pg / ml; # 3 is 100 pg / ml; # 4 is 25 pg / ml; # 5 was 0 pg / ml.

(3) Preparation of Goat anti-rabbit conjugated alkaline phophatase

Enzyme diluents were mixed with vortex and 6 ml were taken, placed in lyophilized chlorine anti-rabbit bound alkaline phosphate and mixed with a rotator.

(4) Preparation of Color reagent

Because the indicator is sensitive to temperature, it was stored before use at 4 ° C. The indicator A and the indicator B were mixed with 1: 1 before use.

(5) Selecting appropriate macrophage condition medium

To analyze TNF-α in macrophage conditioned media, the optimal macrophage conditioned medium volume to be added to 96 wells was determined. That is, 25 μl and 100 μl of macrophage condition medium were taken and reacted with ELISA precoated 96 well fot anti-mouse TNF-α (monocloned), which resulted in the clearest results when 100 μl of medium was administered. As per, TNF-α was then analyzed using 100 μl macrophage condition medium.

(B) TNF-α

Five kinds of TNF-α standard solutions prepared above were added to 100 μl of 96 wells coated with rat anti-mouse TNF-α, and 100 μl macrophage condition medium was added to each well.

In addition, since the antibody-antigen reaction should be performed in neutral, the pH was neutralized to about 7.5 by further adding 100 µl assay dilution to the alkaline alkaline macrophage condition medium.

25 ml of the primary antibody rabbit anti-mouse TNF-α was added to the 96 wells, mixed, incubated at room temperature for 4 hours, and washed 5 times with washing buffer (250 μl / well). Then, 50 μl of a second antibody, goat anti-rabbit mixed alkaline phosphate, was added to the 96 wells, incubated for 45 minutes, washed five times with washing buffer (250 μl / well), and then dried in vacuo. Carefully removed.

200 μl of the indicator was added to the 96 well, and when the red color was distinguished for each well, 50 μl of the stop solution was added dropwise, the reaction of the indicator was fixed, and measured by ELISA reader at 490 nm, and measured by TNF-α. The amount of was analyzed.

There are two types of TNF (tumor necrosis factor), mainly TNF-α is produced in monocytes or macrophages, and TNF-β is produced in lymphocytes. TNF-α is an inflammation mediated cytokine, either in vivo or by interacting with a cytokine such as INF-γ to induce tumor necrosis factor or induce host resistance to microbial infection. However, for radiation or severe stimuli, rapid overproduction has a very detrimental effect on the living body. TNF-α can induce iNOS expression in a wide range of tissues, organs, tumors, cell lines, and the like.

After incubating with the medicinal plant extract of the present invention added to the raw 264.7 cells, RNA was extracted from the cells, RT-PCR and ELISA were performed to analyze mRNA expression levels of cytokines, as shown in FIG. , The expression level of cytokine was decreased with the increase of the concentration of the extract of strengthened wormwood, mRNA expression level also showed the same trend. Therefore, when 100 μg / ml of the extract of Fortified Mugwort was added, there was a marked decrease in the expression level of mRNA.

On the other hand, T cells are divided into Th1 and Th2 cells, and INF-γ is expressed in Th1 lymphocytes, which is one of many cytokines that cause inflammation. INF- [gamma] is produced by expression of STAT1 and is known to promote inflammation. Protein expression of STAT1, p-STAT1, and INF-γ was analyzed in inflammatory bowel disease animal model, but the expression level of STAT1, p-STAT1, and INF-γ was increased in DSS group, whereas expression level was increased in wormwood group. In the DSS + enhanced wormwood group, it was confirmed that the expression level was reduced compared to the DSS group. The results are shown in FIGS. 6 and 7.

In conclusion, by identifying the decrease of iNOS, TNF-α, COX-2, IFN-γ, STAT1, and p-STAT1 expression levels by the extract of Fortified Mugwort of the present invention, inflammation of macrophages such as Raw 264.7 cells may have been induced. At that time, it was found that the strengthening wormwood extract of the present invention is effective in inhibiting inflammation. Therefore, the extract of the strengthening wormwood of the present invention was observed that the excellent anti-inflammatory effect, and confirmed that it can be used clinically as an inflammatory bowel disease and inflammation prevention agent.

Hereinafter, an example of the preparation of a pharmaceutical composition comprising the extract of the present invention, but the present invention is not intended to limit it, it is intended to explain in detail only.

Formulation Example 1.

Powdered wormwood extract of the present invention was mixed with the following components, and the injection was prepared by sterilization by filling in a 2 ml ampoule according to a conventional preparation of injectables.

Fortified Wormwood Extract 100mg

Sodium metabisulfite 3.0 mg

Methine paraben 0.8 mg

Propylparaben 0.1 mg

0.1 mg sterile distilled water for injection

Formulation Example 2.

The extract of Fortified Mugwort of the present invention was mixed with the following ingredients and tableted according to a conventional manufacturing process of the tablet to prepare a tablet containing the same.

Fortified Wormwood Extract 200 mg

Lactose 100 mg

Starch 100 mg

Magnesium stearate appropriate amount

Formulation Example 3.

The extract of Fortified Mugwort of the present invention was mixed with the following ingredients and filled into gelatin capsules according to the conventional preparation of capsules to prepare capsules containing the same.

Fortified Wormwood Extract 200 mg

Lactose 100 mg

Starch 100 mg

Talc 2 mg

Magnesium stearate appropriate amount

Formulation Example 4.

Powdered wormwood extract of the present invention was mixed with the following components and filled into a 100 ml brown bottle according to a conventional process for preparing a liquid to sterilize to prepare a liquid containing it.

Fortified wormwood extract 1,000 mg

20 mg of sugar

Isomerized sugar 20 mg

Lemon flavor

100 ml of whole purified water addition

As described above, the present invention relates to an extract of fortified wormwood having anti-inflammatory activity, a method for preparing the same, and a pharmaceutical composition for an anti-inflammatory agent comprising the fortified wormwood as an active ingredient, the fortified wormwood extract of the present invention is used for inflammatory bowel disease and It can be effectively used for preventing and treating cellular inflammation.

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Claims (2)

Fortified wormwood with anti-inflammatory effect ( Artemisa spp ) pharmaceutical composition for the prevention and treatment of inflammatory bowel disease and cellular inflammation comprising the extract as an active ingredient. The method of claim 1, The fortified wormwood extract is a pharmaceutical composition for the prevention and treatment of inflammatory bowel disease and cell inflammation, characterized in that used as an anti-inflammatory agent.

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