KR101395628B1 - Pharmaceutical composition for preventing and treating acute colitis disease containing the extract of rumex aquaticus herba as active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the prophylaxis and treatment of acute colitis disease containing an extract of Rumex Aquaticus Herba as an active ingredient.
The pharmaceutical composition containing the bean leaf extract of the present invention as an active ingredient not only improves the clinical deterioration of colitis induced in animal models of TNBS-induced acute colitis, but also inhibits NO production, TNF-α , The MPO activity, and the oxidative stress, as well as the effective action of the antioxidant (GSH) reduced by reactive oxygen species (ROS). Furthermore, the pharmaceutical composition containing the extract of the natural mother liquor can be usefully used for the prevention and treatment of acute colitis disease with minimal side effects.

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition for the prevention and treatment of acute colitis disease containing an extract of Bacillus subtilis as an active ingredient. BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a pharmaceutical composition for the prevention and treatment of acute colitis,

The present invention relates to a pharmaceutical composition for the prevention and treatment of acute colitis disease comprising an extract of Rumex Aquaticus Herba as an active ingredient, and more particularly to a pharmaceutical composition for the prevention and treatment of acute colitis caused by 2,4,6-trinitrobenzenesulfonic acid (TNBS) In addition to improving the clinical deterioration of colitis induced by bovine lobe extract in animal models of acute colitis, the expression of NO, TNF-a, MPO activity and lipid peroxidation due to oxidative stress ( Rumex Aquaticus Herba ) extract, which is effective for the recovery of antioxidant (GSH) reduced by active oxygen species (ROS), and to a pharmaceutical composition for the prevention and treatment of acute colitis disease.

Inflammatory bowel disease is one of the immune-mediated chronic bowel diseases including Crohn's disease and ulcerative colitis.

The inflammatory bowel disease causes rectal bleeding and diarrheal epithelial barrier breakdown, and mucosal ulcer is formed. In particular, chronic ulcerative colitis can develop into colon cancer. Currently, medical treatment of inflammatory bowel disease has been developed by medical technology, but no definitive, fundamental treatment method has been proposed yet.

For example, the treatment of chronic ulcerative colitis includes anti-inflammatory therapy with 5-aminosalicylic acid and glucocorticoids or immunotherapy with azathioprine, mercaptopurines and cyclosporin However, these drugs have serious side effects and a high recurrence rate.

The causes of etiology of ulcer and colitis have not been elucidated yet, but they are generally accepted to be caused by infiltration of neutrophils and overexpression of pro-inflammatory mediators such as reactive oxygen species and cytokines.

In general, as a model of Crohn's disease and ulcerative colitis, 1,3,5-trinitrobenzenesulfonic acid (TNBS) causes severe chronic inflammation and is used in the rectum.

The TNBS-induced colitis model leads to an abnormal immune response that is primarily mediated by the first activated mast cells and leukocytes, particularly neutrophils, which increase reactive oxygen and nitrogen.

From the TNBS-induced colitis model, we investigate tumor necrosis factor (TNF-a), which is known to be involved in inflammatory bowel disease.

Active oxygen produced and released by immune cells plays an important role in the pathology of ulcerative colitis, and oxidative stress and consequently lipid peroxidation exacerbates the free radical chain reaction. Obstruct the integrity of the intestinal mucosal barrier, and activate inflammation mediators. For example, the degree of colonization of malondialdehyde (MDA), a major component of lipid peroxides, has increased the incidence of inflammatory bowel disease in animal studies.

Thus, MDA levels are often used as oxidative damage indicators and as free radical-induced lipid peroxidation markers. Increased myeloperoxidase (MPO) activity and subsequent production of nitric oxide (NO) have been demonstrated in inflammatory colon and disease progression in biopsies.

Nitric oxide (NO) reacts with superoxide to inhibit the enzymatic action of the major role in electron transport in mitochondria, inducing the production of peroxynitrite, a deadly cytotoxic oxidant, and inducing iNOS expression through NF-κB And can decrease the expression of prostacyclin synthase-protein. Thus, excessive nitric oxide (NO) production accelerates inflammatory bowel disease.

Flavonoids are polyphenolic compounds that are recognized for their anti-inflammatory, neuroprotective, chemopreventive, immune and gastro-protective effects. However, the efficacy of flavonoids for the inflammatory response of models with acute and chronic colonic injury is rarely reported.

Among them, Artemisia asiatica is frequently used in civilian applications such as abdominal pain, bloody stool, internal bleeding and gynecological diseases, and it is also called "lye".

It has been reported that the extract of liana, which is an ethanol extract of dried powder of mugwort, has a protective effect against mucosa due to antioxidant action in esophagus and stomach.

As another example, Rumex Aquaticus Herba is a perennial herb that is distributed in Korea (Gyeongbuk, Hamnam), Japan, Europe, and North America, and is called Shigeru, Songgi, In Korea and China, its roots are called Yangjingun, and its leaves are called Yangjiyap. Known ingredients in Rumex species plants include, but are not limited to, 3-acetyl-2-methyl-1,5-dihydroxy-2,3-epoxynaphthoquinol in Rumex japonicus -methyl-1, 5-dihydroxy- 2, 3-epoxynaphthoquinol) (Zee OP, et al., Arch. Pharm. Res., Aug 21 (4) to, pp.385-6, 1998), No. rhubarb (Rumexgmelini ), 1-O-? -D-glucopyranosylmodine ( Chung Kuo Chung Yao Tsa Chih , Dec 21 (12) , pp.741-2, 1996 ) was isolated from the root (1β6) galactoside, quercetin-3-glycosyl (1β4) galactoside, emodine (Hasan A, et .. al, Phytochemistry, Jul 39 (5), pp.1211-3, 1995) the plants dongsok (Rumex luminastrum) Cri platelets (chrysophan) acid, in emodine (emodin), four podin (in nepodin; 2-acetyl- 1, 8-dihydroxy-3- methyl nahthalene) (Abe el-Fattah H., et al., Acta Pharm Hung, may 64 (3), pp.83-5, 1994) and the like have been reported, rivers It is contained per ingredients. It is used as a medicine in insect, diarrhea, fever, swelling, jaundice, constipation, The rooted water has the effect of stopping uterine bleeding and nose, etc. It also has effects on vomiting, diarrhea and constipation, it has excellent sterilizing effect and can be used for cancer treatment, and it has significant therapeutic effect on myeloid leukemia and lymphocytic leukemia have. In addition, it has a function to lower fever and sweat, so it is effective for cold, pulmonary tuberculosis and cough. It has subcutaneous, antipyretic, hypothermic, and bile secretion-promoting actions and decreases the blood coagulation time because it reduces the permeability of capillaries. It has a broad antibacterial action and inhibits the growth of staphylococcus, gonorrhea, diphtheria, typhoid, and alien bacteria. In addition, diuretic and liver function protection appears.

Thus, the present inventors investigated the efficacy of flavonoid compounds in acute and chronic intestinal damage induced models, confirming the anti-inflammatory effect of colchicine in animal models of acute colitis induced by TNBS in flavonoid extracts , Thereby completing the present invention.

It is an object of the present invention to provide a pharmaceutical composition for the prevention and treatment of acute colitis disease containing an extract of Rumex Aquaticus Herba as an active ingredient.

It is another object of the present invention to provide a pharmaceutical composition containing a quercetin-3-beta-di-glucuronopyranoside compound component isolated from bean leaf extract useful for the prevention and treatment of acute colitis disease.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of acute colitis disease containing an extract of Rumex Aquaticus Herba as an active ingredient.

The bean leaf extract of the present invention is an extract which is soluble in water, a C ₁ to C ₄ lower alcohol solvent or a mixed solvent thereof. More preferably, the pharmaceutical composition of the present invention is quercetin-3-beta-di - glucuronopyranoside compound component.

At this time, the extract of Rumex Aquaticus Herba of the present invention is preferably contained in an amount of 30 to 100 mg / kg.

INDUSTRIAL APPLICABILITY The present invention can be provided as a pharmaceutical composition for the prevention and treatment of an acute colitis disease containing an extract of bamboo leaf as an active ingredient, because the extract of bamboo leaves is effective for anti-inflammatory activity in an animal model of TNBS-induced acute colitis.

In addition, the pharmaceutical composition containing the quercetin-3-beta-di-glucuronopyranoside compound component isolated from the bean leaf extract of the present invention is extracted from a natural component and provided with biostability, And exhibit similar or superior activity to the drug, they can be provided as pharmaceutical compositions with minimized side effects.

Brief Description of the Drawings Fig. 1 shows the effect of the extract of two-leaf extract against colon injury in an animal model of acute colitis induced by TNBS of the present invention,
FIG. 2 shows the results of measuring the effect of the extract of bovine leaf extract on the weight per the colonic length in an animal model of TNBS-induced acute colitis of the present invention,
FIG. 3 shows the results of observing the effects of MPO activity per the colonic length on the administration of the extract of bovine lobe extract in an animal model of TNBS-induced acute colitis of the present invention,
FIG. 4 shows the results of observing the effect of the extract of bovine lobe extract on NO production in an animal model of TNBS-induced acute colitis of the present invention,
FIG. 5 shows the results of observing the effect of TNF-α expression on the animal model of TNBS-induced acute colitis according to the present invention,
FIG. 6 shows the results of observing the effect of the extract of bovine leaf extract on the GSH change in the animal model of TNBS-induced acute colitis of the present invention,
FIG. 7 shows the results of observing the effect of the extract of bovine lobe extract on the change of MDA concentration in an animal model of TNBS-induced acute colitis of the present invention.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for the prevention and treatment of acute colitis disease which contains Rumex Aquaticus Herba extract as an active ingredient.

The bean leaf extract of the present invention is an extract which is soluble in water, a lower alcohol solvent of C ₁ to C ₄ or a mixed solvent thereof, more specifically, water of Rumex Aquaticus Herba leaf or C ₁ to C ₄ lower alcohol Is removed by using a non-polar solvent such as chloroform, ether or methylene chloride, and the sugar and the inorganic salt are removed using a non-ion exchange resin. Recrystallization of the resulting 40% Quercetin-3 -? - D-glucuronopyranoside compound component.

Formula 1

Accordingly, the present invention provides a pharmaceutical composition containing the quercetin-3-beta-di-glucuronopyranoside compound component represented by the formula (1) isolated from the above-mentioned two-leaf extract.

The present invention relates to a pharmaceutical composition for the prevention and treatment of acute colitis disease containing an extract of Rumex Aquaticus Herba as an active ingredient, and more particularly to a pharmaceutical composition for preventing and treating acute colitis caused by TNBS-induced acute colitis, Not only improves the clinical deterioration of colitis, but also improves the degree of lipid peroxidation due to NO production, TNF-a expression, MPO activity, and oxidative stress, which affect the inflammatory response, and decreases by ROS (GSH). ≪ / RTI >

Specifically, the pharmaceutical composition for preventing and treating acute colitis disease containing the extract of Rumex Aquaticus Herba of the present invention as an active ingredient improves the clinical deterioration of induced colitis [ Fig. 1 ], tissue edema As a result of observing the ratio of the weight per length of the colon observed as an index of infiltration of the inflammatory substance, a low weight per length of the colon compared to the conventional drug for treating colitis can be confirmed ( Fig. 2 ).

In order to confirm the pharmaceutical efficacy of the bean leaf extract of the present invention, the MPO activity was observed in a dose-dependent manner when the extract of the bean leaf was administered ( FIG. 3 ).

In addition, observation of the effect on NO production affecting the inflammatory response, as administered a dose-dependent manner of yangjeyeop extract of NO generated is decreased, and [4], yangjeyeop in the observation of a TNF-α expression involved in neutrophil activation The expression of TNF-α is dose-dependently reduced upon administration of the extract ( FIG. 5 ).

In addition, in order to verify the pharmacological efficacy of the bean leaf extract of the present invention, the GSH level oxidized by GST (Glutathione S-transferases), which plays an important role in the defense mechanism of the human body against cancer-causing chemical species or other toxins Changes were observed. As a result, it was confirmed that the administration of the extract of bovine L. extract increased total GSH production and increased dose-dependently upon administration ( Fig. 6 ).

In addition, malondialdehyde (MDA), a lipid peroxide produced by the reaction between reactive oxygen species (ROS) and lipid increased during inflammation, decreased MDA levels in a dose-dependent manner , And this behavior is lower than that of conventional drugs for treatment of colitis. The extract of the present invention is not only useful for the prevention and treatment of colitis, but also provides a possibility of substituting for conventional antiinflammatory drugs ( Fig. 7 ) .

From these results, the extract of Rumex Aquaticus Herba of the present invention is preferably contained in an amount of 30 to 100 mg / kg in order to optimize the expression of the anti-inflammatory effect.

The pharmaceutical composition containing the bean leaf extract of the present invention may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories, Can be used.

Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition containing the extract of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, Calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

When the pharmaceutical composition containing the extract of the present invention is formulated, it is prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, sucrose), lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used.

Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

The preparation for parenteral administration may include a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a freeze-dried preparation, and a suppository. Examples of the non-aqueous solution and the suspending agent include propylene glycol, polyethylene glycol, Vegetable oil, injectable esters such as ethyl oleate, and the like can be used. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The preferred dosage of the composition of the present invention varies depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration and the period of time, but can be appropriately selected by those skilled in the art. However, for the desired effect, the extract or fraction of the present invention is preferably administered at 0.5 g / kg to 5 g / kg per day, preferably 1 g / kg to 3 g / kg per day. The administration may be carried out once a day or divided into several doses. Accordingly, the dosage is not limited in any way to the scope of the present invention.

The composition of the present invention is administered orally to mammals such as rats, mice, livestock, humans, etc., but may be administered by various routes of administration known in the art. For example, by oral, rectal or intravenous, intramuscular, subcutaneous, intramural or intracerebroventricular injection.

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

The present invention is intended to more specifically illustrate the present invention, and the scope of the present invention is not limited to these embodiments.

≪ Example 1 >

The herb extracts from herbivorous leaves provided in the medicinal botany of the College of Pharmacy, Chung - Ang University were dissolved in distilled water for animal experiments.

Male Sprague-Dawley (SD) male rats (Samutako Bio), weighing 180 to 200 g, were raised in a cage maintained at a temperature of 24-25 ° C and a relative humidity of 70-75% The food was supplied with a diet (provided by Samtako Bio Co., Ltd.).

Thereafter, the experimental rats were fasted for 24 hours before inducing colitis, and water was supplied in sufficient quantities. Animals were divided into six groups. Urethane (0.5 g / ㎖) was intraperitoneally administered and a medical polyurethane cannula (2 mm in outer diameter) was inserted into the anus. 120 mg / ml of TNBS was dissolved in ethanol (50% v / v) and 0.25 ml of TNBS solution was intravenously injected via a cannula to induce colitis. Then, the head of the experimental mouse was held down for 2-3 minutes to prevent leakage. TNBS was injected into the large intestine to induce colitis, and oral administration of 30 mg / kg of the extract of lamb extract was administered 48 hours, 24 hours, 1 hour before and 24 hours after induction of colitis. The animals were all sacrificed 48 hours after induction of colitis. As the negative control group, the same amount of 0.9% salt was administered in the colonic instead of the TNBS solution. The behavioral and weight changes and stool consistency of all experimental mice were monitored daily. At this time, it is preferable that the amount of 1,3,5-trinitrobenzenesulfonic acid (TNBS), sodium carboxymethyl cellulose (CMC-Na), 5-aminosalicylic acid (5-AS), EDTA, EGTA, urethane, hexadecyltrimethylammonium Bromide (HETAB), hydrogen peroxide, and dimethyl formaldehyde were purchased from Sigma.

≪ Example 2 >

The same procedure as in Example 1 was carried out except that 50 mg / kg of the bean leaf extract used in Example 1 was orally administered.

≪ Example 3 >

The same procedure as in Example 1 was carried out except that 100 mg / kg of the bean leaf extract used in Example 1 was orally administered.

<Example 4> Separation of compounds from extracts of two leaves

The extract obtained by adding 3-fold amount of 80% methanol to 4 kg of the baby leaves provided in the medicinal botany of the College of Pharmacy, Chung-Ang University and refluxing for 3 hours at a temperature of 40-60 ° C was diluted with a reduced pressure concentrator (Eyela, FD-5N Japan) The obtained extract was dissolved in 500 ml of distilled water and suspended. The resulting extract was separated by filtration through a filter paper into a water-insoluble layer and a water-soluble layer, and then the water-insoluble layer was removed. Of chloroform.

The water-soluble portion after the degreasing process was removed by chloroform solvent using a vacuum concentrator, adsorbed with nonionic resin (XAD-2) for column chromatography, and then eluted with 4000 ml of distilled water to remove sugar and inorganic salts. 20% , 40% methanol, 60% methanol, 80% methanol and 100% methanol sequentially to obtain 6 g of flavonoid-containing fraction eluted with 40% methanol. 6 g of the flavonoid-containing fraction was subjected to recrystallization to identify and confirm pure 2.1 g of quercetin-3-beta-di-glucuronopyranoside (QGC) compound [mp: 193-195 ° C, (-) FAB MS (m / z): 477 [MH] -, 301 [M- (GlcUA = H)] - 1H-NMR (300 MHz, DMSO-d6); (1H, d, J = 2.4, 6.3 Hz, H-6 '), 7.51 (1H, s, H-2'), 6.82 (1H, d, J = 7.2 Hz, GluUA H), 6.39 (1H, d, J = 2.1 Hz,

13 C-NMR (75 MHz, DMSO-d 6); 156.4 (C-2), 133.3 (C-3), 177.4 (C-4), 161.4 (C-5), 98.9 (C-9), 104.0 (C-10), 121.0 (C-1 '),

115.3 (C-2 '), 145.1 (C-3'), 148.8 (C-4 '), 116.2 (GluUA: C-2), 76.1 (GluUA: C-3), 71.4 (GluUA: C-4), 75.9 (GluUA: C-5), 170.0 (GluUA: C-6).

&Lt; Comparative Example 1 &

The procedure of Example 1 was repeated, except that 5-aminosalicylic acid (5-AS), normally used in the treatment of colitis, was orally administered at 25 ml / kg in place of the bean leaf extract used in Example 1.

&Lt; Comparative Example 2 &

The procedure of Example 1 was repeated, except that 10 mg / kg of the lentil extract used in Example 1 was orally administered.

&Lt; Experimental Example 1 >

If one with respect to the animal model of acute colitis induced by TNBS, oral administration of a yangjeyeop extract 30mg / kg conducted in Example 1 were shown in Figure 1 to determine the degree of mucosal damage of the colon by the naked eye.

FIG. 1 shows the effect of the extract of the present invention on colon damage in an animal model of TNBS-induced acute colitis of the present invention, wherein A is a normal group, B is a group treated with TNBS, The results of this study are as follows.

From the above results, the inventive bean leaf extract was administered, and the ulcer reduction effect on acute colitis was visually confirmed.

Table 1 below shows the clinical changes according to the drug administered in an animal model of TNBS-induced acute colitis.
[Table 1]

As a result of the above Table 1, Example 1 and Example 2, which were orally administered 30 mg / kg and 50 mg / kg of the extracts of both leaves from animal models of TNBS-induced acute colitis, showed that 5-aminosalicylic acid (5-AS ), Which is comparable to Comparative Example 1, which is a drug administration group, and showed low tissue adhesion.

Particularly, in Example 3 in which 100 mg / kg of bovine leaf extract was orally administered, the body weight change range was the lowest, the tissue adhesion rate was significantly lowered, and side effects such as diarrhea were low.

On the other hand, in the case of Comparative Example 2 in which 10 mg / kg of bovine leaf extract was administered, the inflammation-inhibitory effect was not expressed, and thus, it was excluded from the experimental group.

&Lt; Experimental Example 2 > Evaluation of effect on weight per length of bowel

For animal models of TNBS-induced acute colitis, 48 hours after induction of colitis, the cervical spine of the mouse was decapitated and the colon was resected to about 10 cm, and the weight per length was measured.

Figure 2 shows the results of measuring the effect of the substances administered in Examples 1 to 3 and Comparative Example 1 on the weight per length of colon in animal models of TNBS-induced acute colitis of the present invention.

The weight ratio of the large intestine to the length of the colon is an index of tissue edema and infiltration of inflammatory substances. From the results of Examples 1 to 3, the weight-dependent weight loss was reduced in the administration of the extract of the two- Was lower than the results of Comparative Example 1 in which &lt; RTI ID = 0.0 &gt; Therefore, it is supported that, when 30-100 mg / kg of the extract of the present invention extract of bean curd, is administered, a substance used for the treatment of common colitis can be used alternatively.

<Experimental Example 3> Anti-inflammatory effect

For animal models of TNBS-induced acute colitis, the anti-inflammatory effects of the substances administered in Examples 1 to 3 and Comparative Example 1 were observed in the model.

1. MPO activity

MPO activity assay was performed by modification from the preceding method by Grisham et al. FIG. 3 shows the results of observing the myeloperoxidase (MPO) activity according to the administration of the extract of bovine bark extract in an animal model of TNBS-induced acute colitis of the present invention.

The activity of MPO, an enzyme affecting cytotoxicity through the production of HOCl, was rapidly elevated after TNBS injection compared to the control group, and this was used as an animal model of TNBS-induced acute colitis. Thereafter, MPO activity was observed according to the substances administered in Examples 1 to 3 and Comparative Example 1. As a result, it was confirmed that the MPO activity tended to decrease in a dose-dependent manner upon administration of the extract of bovine lobe extract. Particularly, in the case of Example 3, similar results are obtained as compared with Comparative Example 1, thereby supporting that the natural herbal extract of the present invention can be used for treating colitis.

2. Effect on NO production

FIG. 4 shows the results of observing the effect of the extract of the present invention on the production of NO in the animal model of TNBS-induced acute colitis according to the present invention.

As a result, the degree of NO production, which directly affects the inflammatory response to intestinal epithelial cell injury, neutrophil activation, and vasodilation in the tissues, was significantly increased after injection of TNBS into the colon.

However, in the case of Examples 1 to 3 in which the bean leaf extract of the present invention was administered, NO production was dose-dependently decreased upon administration.

3. Effect on TNF-α expression

FIG. 5 shows the effect of TNBS-induced expression of TNBS on the expression of TNF-α associated with neutrophil activation in an animal model of TNBS-induced acute colitis of the present invention. Compared to the control group.

However, as a result of administration of the extract of the present invention, the expression of TNF-α was decreased in a dose-dependent manner.

&Lt; Experimental Example 4 >

(GSH) induced by GST (Glutathione S-transferases) in the presence of toxic lipid or hydrogen peroxide upon inflammation.

The GST plays an important role in the defense mechanism of the human body against cancer-causing chemical species or other toxins. It removes the reactive oxygen species (ROS) resulting from oxidative stress or regenerates sulfur protein Perform a wide range of functions in the cell.

FIG. 6 shows the effect of the extract of the present invention on the change of GSH in the animal model of TNBS-induced acute colitis according to the present invention. As a result, the GSH level in the colon was significantly Respectively.

However, in the case of Examples 1 to 3 in which the extracts of both plants were administered, the total GSH production was dose-dependently increased, and the results were comparable to those of Comparative Example 1 administered with a commercial colitis therapeutic.

From the above results, it was confirmed that the extract of the present invention improved the degree of lipid peroxidation due to oxidative stress and had a beneficial effect on the recovery of GSH, which is an antioxidant reduced by ROS.

&Lt; Experimental Example 5 > Effect on MDA change

Reactive oxygen species (ROS) upon inflammation react with lipids to produce malondialdehyde (MDA), a lipid peroxide. Thus, in the animal model of TNBS-induced acute colitis of the present invention, TNBS The concentration of MDA increases sharply.

FIG. 7 shows the results of observing the effect of the extract of the present invention on the change of MDA concentration in the animal model of TNBS-induced acute colitis of the present invention. In Examples 1 to 3, The MDA concentration level was decreased in a dose-dependent manner, and the decrease level was observed to be lower than that of Comparative Example 1, which is used for the treatment of normal colitis. From the above results, it can be concluded that the bean leaf extract of the present invention is a natural pharmaceutical composition and has superior antiinflammatory effect than a conventional drug for treating colitis, and thus can be used as an alternative.

From the above results, the animal model of TNBS-induced acute colitis of the present invention causes a sharp increase of active oxygen species, hydrogen peroxide, and oxygen radicals, which shows an inflammatory reaction such as neutrophil activation.

Thus, when the extract of the present invention is administered, not only the clinical deterioration of the induced colitis is improved but also the production of NO, the expression of TNF-α associated with neutrophil activation, and the production of HOCl We have confirmed the results that have beneficial effects on the recovery of GSH, an antioxidant reduced by reactive oxygen species (ROS), while improving lipid peroxidation due to MPO activity and oxidative stress, which are cytotoxic enzymes.

&Lt; Experimental Example 6 > Acute toxicity test for oral administration to experimental mice

The following experiments were conducted to investigate the acute toxicity of the extracts.

Oral dose toxicity studies were performed on 5 mice and 6 mice of 10 mice in each group for experimental mice weighing 20 g. The herb extracts were administered once at a dose of 2,000 mg / kg for 7 days. After the administration of the test substance, the animals were observed for death, clinical symptoms and weight changes, and hematological tests and blood biochemical tests were carried out. As a result of the test, there was no clinically symptomatic symptom in all the animals to which the test substance was administered, no mortality was observed, and no toxic change was observed in weight change, blood test, blood biochemical test or autopsy.

All of the tested extracts showed no toxicity in experimental mice and the oral dose of LD ₅₀ was more than 2,000 mg / kg, which is biostable and safe to administer.

As described above, the present invention provides a pharmaceutical composition in which an extract of bovine leaf extract is effective for anti-inflammatory activity in an animal model of acute colitis induced by TNBS.

The present invention provides a pharmaceutical composition containing a quercetin-3-beta-di-glucuronopyranoside compound component isolated from the above-mentioned two-leaf extract.

The pharmaceutical composition of the present invention is extracted from a natural component and is supported by biostability. It not only improves the clinical deterioration of induced colitis, but also enhances NO production, TNF-a expression, MPO activity, (GSH) reduced by active oxygen species (ROS), and thus can be utilized as a pharmaceutical composition with minimized adverse effects of natural ingredients.

While the invention has been shown and described with reference to certain exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (4)

The extract of Lambskin extract contains quercetin-3-beta-D-glucuronopyranoside represented by the following formula (1) A pharmaceutical composition for the prevention and treatment of acute colitis disease which comprises:
[Chemical Formula 1]

delete delete The pharmaceutical composition for preventing and treating acute colitis disease according to claim 1, wherein the extract of Rumex Aquaticus Herba is contained in an amount of 30 to 100 mg / kg.

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