KR101457969B1 - Pharmaceutical composition comprising leaf extract of Abutilon theophrasti Medicus - Google Patents

Abstract

The present invention relates to a pharmaceutical composition containing a leafy leaf extract as an active ingredient and a health functional food. More specifically, the pharmaceutical composition and the health functional food have anti-inflammatory, antioxidant and anti-arteriosclerotic activity.

Description

[0001] The present invention relates to a pharmaceutical composition comprising leaf extract of Abutilon theophrasti Medicus,

The present invention relates to a pharmaceutical composition containing a leafy leaf extract as an active ingredient and a health functional food.

It is a perennial plant belonging to the family Eucalyptus. Its stem is cylindrical and 1.5m high. Yellow flowers bloom from July to August in the stem of the stem at the end of the stem, and the fruit forms an eggplant (朔 果). It is native to India and cultivated in Korea, Japan and China. The stem skin is used to make rope and saddle with fiber, and the residue is used as paper raw material. The seeds are used as medicinal herbs because they are well - suited for diarrhea and are used mainly for the treatment of edema and constipation during pregnancy. The leaf is 2 ~ 15 cm long with petiole and the leaf is round in heart, acutely pointed and dull serrate on the edge. Star-shaped hairs on both front and back. The seeds are used as herbal medicines, but the leaves are rarely used as medicinal or pharmaceutical compositions.

Antioxidation refers to the prevention of many oxidative effects in the body. Oxidation refers to the phenomenon that when various kinds of unsaturated fatty acids present in the body are oxidized by oxygen, a substance called lipid peroxidation is produced, and when it is increased, aging phenomenon is promoted. Human diseases and aging can be classified into _two groups: O ₂ ^- (superoxide), NO ₂ (nitric oxide), NO ₂ · (nitrogen dioxide), OH · (hydroxyl), ROO · (proxyl) ), NO ₃ · (peroxynitrite) radicals and the like. In vivo, as a means to protect the living body from free radicals such as superoxide dismutase (SOD), catalase (CAT catalase, and glutathione peroxidase (GSHpx), and vitamin E and glutathione to protect the body from oxidative damage by eliminating free radicals. However, when such an antioxidant defense system declines to such an extent that free radical formation can not be controlled, the oxidative stress and damage of the tissue are promoted, and the excess free radicals and lipid peroxides produced in the body are oxidized by oxidative stress such as protein oxidation and DNA damage . Therefore, many synthetic or natural antioxidants have been developed for the reduction of oxidative stress. However, synthetic antioxidants have a problem in safety. In recent years, a safe and antioxidant natural antioxidant in various herbal and edible plant extracts We are making efforts to develop.

Inflammation is an expression of various types of infection and pre-in vivo defense against irritants in metabolites in vivo, and various chemical mediators are involved in the mechanism of inflammation expression, Complex. The most powerful anti-inflammatory drug developed by the first-class drug is steroids. However, long-term use is accompanied by side effects. Therefore, steroids are surprisingly effective when used for the first time, and they cause symptoms to disappear completely but only for a short time. Symptoms of Steroids appear again with stopping the use of steroids. . Side effects of steroids include side effects such as round polyhedronic face, retention of fluid, adrenal suppression, increased susceptibility to infection, and other psychoses, cataracts, glaucoma, peptic ulcer, delayed wound healing, I have. Therefore, it is necessary to study a substance which can be safely ingested safely without a separate purification process, has an effect of inhibiting inflammation, and can be easily used for food, as a medicament containing a natural plant extract as an active ingredient.

Atherosclerosis is a disease in which arteries are hardened by environmental factors such as dietary factors related to lipid metabolism and eating habits, smoking, and lack of exercise, which causes circulatory diseases such as heart diseases and cerebrovascular diseases. The hypothesis about the early development of atherosclerosis is a response-to-injury hypothesis that can be used to detect vascular endothelial cells by genetic mutations, peroxides, hypertension, diabetes, increased plasma homocysteine concentration, It is in a state of malfunction that does not maintain normal homeostasis. When the endothelial cells become dysfunctional, the cell adhesion substance is largely expressed and the cell permeability is increased, so that the permeability to the adherence and tissue of the immune cells, platelets, and lipids of the blood is increased, and the inflammatory mediators and growth factor secretion Inflammatory response to arteriosclerotic lesions occur and develop. At this time, low density lipoprotein (LDL) in the blood is converted to LDL (modified-LDL) due to oxidation, glucose binding, aggregation, binding of glycoprotein and the like, and they are stimulated by vascular endothelial cells and smooth muscle Causing damage. As a result, when expression of vascular cell adhesion molecule-1 (VCAM-1) and release of inflammatory mediators of inflammatory cells are promoted, LDL is introduced and accumulated under endothelial cells, LDL and oxidized MLDL re-invade the immune cells, such as macrophages and T lymphocytes, and promote the inflammatory response of the lesions. Thereafter, the lesion becomes necrotic due to the action of hydrolytic enzymes, inflammatory mediators, and growth factors released from the macrophages or lymphocytes infiltrated into the lesions. The necrosis of the lesion leads to the infiltration of mononuclear cells, migration and differentiation of smooth muscle, Through the repetitive process such as formation of the tissue, the lesion tissue develops into a fibrous lesion of a complicated structure covered with fibrous necrotic tissue composed of MLDL nucleus. The thrombus is formed from the developed lesion tissue and the artery is hardened, Circulatory diseases will appear. Therefore, it is urgently required to develop an anti-arteriosclerosis agent which inhibits the oxidative action of LDL to prevent arteriosclerosis, exerts excellent therapeutic effect, and has few side effects.

Accordingly, the present invention provides a natural antioxidant which is safer than an artificially synthesized antioxidant and has an excellent antioxidative effect. Further, the present invention provides an anti-inflammatory agent which is excellent in anti-inflammatory effect and less side effect than an artificially synthesized anti-inflammatory agent. The present invention also provides a pharmaceutical composition for prevention and treatment of atherosclerosis, which has fewer side effects than conventional atherosclerosis treating agents and is excellent in the prevention and treatment of arteriosclerosis.

In order to solve the above problems, the present invention relates to anti-inflammatory, antioxidant and anti-arteriosclerotic activity of a leafy leaf extract, and provides a pharmaceutical composition containing an edible leaf extract as an active ingredient.

According to a preferred embodiment of the present invention, there can be provided a pharmaceutical composition for the prevention and treatment of arteriosclerosis and inflammatory diseases containing an extract of a leafy leaf as an active ingredient.

The present invention can provide a pharmaceutical composition for prevention and treatment of inflammatory diseases, which contains an effective amount of a leafy leaf extract having an excellent anti-inflammatory effect and less side effects than a conventional synthetic anti-inflammatory agent. The present invention also provides a pharmaceutical composition for prevention and treatment of arteriosclerosis, which contains lesser side effects than conventional arteriosclerotic drugs and has an effective effect of preventing and treating arteriosclerosis and having an excellent effect on the treatment of arteriosclerosis. The present invention also provides a health functional food having anti-inflammatory, antioxidant and anti-arteriosclerotic activity, which contains safflower leaf extract as an effective ingredient, which is safer and safer than antioxidative effect of conventional synthetic antioxidants.

FIG. 1 is a graph showing the cell survival rate of Raw 264.7 cells on a leafy leaf extract, and Con: a reference group in which no drug is treated, and LPS: 1 μg / ml of inflammation-induced negative control.
FIG. 2 is a graph showing the inhibitory effect on the peroxynitrite formation of the leafy leaf extract. Neg: Negative control treated with con: sodium phosphate buffer (180); And sin 1: sodium phosphate buffer, followed by 50 μM of SIN-1.
FIG. 3 is a graph showing the inhibitory effect on the formation of nitrogen monoxide of a leafy leaf extract, Con: untreated negative control; Lt; RTI ID = 0.0 > llg / ml < / RTI > of LPS.
FIG. 4 is a graph showing the inhibitory effect on the expression of inflammatory proteins COX-2, iNOS and NF-kB in Raw 264.7 cells, and Con: untreated control group; And LPS: 1 μg / ml of LPS.
5 is a graph showing the LDL oxidation inhibition rate (%) of the leafy leaf extract.

The present invention provides a pharmaceutical composition containing a leafy leaf extract as an active ingredient and a health functional food. In addition, the leaf blade extract of the present invention exhibits anti-inflammatory, antioxidant and anti-arteriosclerotic activity.

The extract comprises extracts extracted with a C ₁ to C ₄ alcohol, preferably methanol or ethanol.

And a pharmaceutical composition characterized in that the target disease of the pharmaceutical composition is an arteriosclerosis or an inflammatory disease.

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

The leafy leaf extract of the present invention is preferably but not limited to be produced by a manufacturing method comprising the following steps.

Abutilon theophrasti Drying and pulverizing the Medicus leaf;

Extracting the dried and pulverized leaves with an extraction solvent; And

And concentrating the extracting solvent contained in the extract by vacuum concentration.

In the above method, the diabetic leaf may be used without limitation such as cultivated or commercially available leaf. The drying is preferably, but not exclusively, reduced-pressure drying, vacuum drying, boiling drying, spray drying or lyophilization. The pulverization is preferably, but not exclusively, pulverized or pulverized.

The extraction solvent is preferably an alcohol solvent. As the alcohol, it is preferable to use a C ₁ to C ₄ alcohol, more preferably to use ethanol or methanol.

As the extraction method for extracting the extract from the above-mentioned diabetic leaf, it is preferable to use accelerated solvent extraction or reflux extraction, but not limited thereto. The extraction temperature is preferably 30 to 70 DEG C, but is not limited thereto.

In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator, but not limited thereto.

The present invention provides a pharmaceutical composition for the prevention and treatment of arteriosclerosis or inflammatory diseases containing an extract of a leafy leaf as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or ameliorating atherosclerosis or inflammatory disease, characterized in that the leafy leaf extract has anti-arteriosclerosis activity by inhibiting the oxidation of low density lipoprotein (LDL).

The present invention provides a pharmaceutical composition for preventing or ameliorating arteriosclerosis or inflammatory diseases, characterized in that the leafy leaf extract has anti-inflammatory activity by inhibiting the expression of iNOS, COX-2 or NF-kB.

The present invention provides a pharmaceutical composition for antioxidants containing an extract of a leafy leaf as an active ingredient.

The antioxidant is a substance that releases electrons to active oxygen and thereby oxidizes itself to prevent oxidation of the gene. The active oxygen is reduced by an antioxidant which is a substance that removes active oxygen, thereby losing carcinogenicity. Therefore, antioxidants are cancer-preventing substances and chemically reducing agents. That is, an antioxidant is a substance that protects the body from oxidation by active oxygen by removing active oxygen.

The present invention provides a pharmaceutical composition for an antioxidant characterized in that the leafy leaf extract has an antioxidative activity by inhibiting the production of peroxynitrite or nitrogen monoxide.

The pharmaceutical composition comprising the leaf blade extract according to the present invention as an active ingredient can be administered orally or parenterally in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, May be formulated in the form of sterile injectable solutions.

More specifically, when formulating the composition, it can be prepared using a diluent or an excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like. 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 and talc may also be used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The amount of the leaf blade extract according to the present invention may vary depending on the age, sex and body weight of the patient, but is generally in the range of 0.01 to 50 mg / Kg, preferably 0.1 to 10 mg / Kg, Can be administered separately. Also, the dosage of the leafy leaf extract may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Accordingly, the dosage is not limited in any way to the scope of the present invention.

The present invention provides a pharmaceutical composition comprising an extract of a leafy leaf as an active ingredient for the prevention and treatment of arteriosclerosis or inflammatory diseases.

The inflammatory diseases include inflammatory diseases such as edema, conjunctivitis, periodontitis, rhinitis, otitis, rhinitis, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, ulcerative colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia Selected from the group consisting of psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendinitis, hay fever, perianal inflammation, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis and various acute and chronic inflammatory diseases But it is not limited thereto.

The present invention provides a health functional food for preventing or ameliorating atherosclerosis or inflammatory disease containing an extract of a leafy leaf as an active ingredient.

The present invention provides a health functional food having an antioxidative activity containing an extract of a leafy leaf as an active ingredient.

The health functional food of the present invention can be used as it is, or can be used with other food or food ingredients, and can be suitably used according to conventional methods.

There is no particular limitation on the kind of health functional food. Examples of the foods to which the leafy leaf extract can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen noodles, gums, ice cream, soups, Alcoholic beverages, and vitamin complexes, and they can be used in the form of pills, powders, granules, infusions, tablets, capsules or beverages, and they include all health functional foods in the conventional sense.

In general, the health functional food composition of the present invention may be added to the food or beverage in an amount of 0.01 to 15% by weight based on the total weight of the food, and 0.02 to 10 g , Preferably 0.3 to 1 g.

The health beverage composition of the present invention is not particularly limited to a liquid ingredient other than the above-mentioned diabetic leaf extract as an essential ingredient in the indicated ratios and may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages . Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; Polysaccharides such as dextrin, cyclodextrins; And sugar alcohols such as xylitol, sorbitol, and erythritol. As natural flavors other than those described above, natural flavors (such as tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin)) and synthetic flavors (saccharin, aspartame, etc.) have.

In addition to the above, the health functional food of the present invention may contain various kinds of nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and heavies (cheese, chocolate etc.), pectic acid and its salts, And salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.

In addition, the health functional foods of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks. These components may be used independently or in combination.

According to another preferred embodiment of the present invention, the alcohol-extracted extract of a leaf of a leaf is selected from the group consisting of cyclooxygenase-2 (COX-2), nitric oxide synthase (iNOS) The present invention can provide a health functional food for preventing or ameliorating atherosclerosis or inflammatory disease, which has an anti-inflammatory activity by inhibiting the expression of nuclear factor kappa B (NF-kB).

According to another preferred embodiment of the present invention, an extract obtained by extracting a leaf of a diabetic rats with an antioxidant activity by inhibiting the production of peroxynitrite (ONOO ^- ) or nitric oxide (NO ^- ), Functional foods can be provided.

According to another preferred embodiment of the present invention, an extract obtained by extracting a leaf of Alaska pollack with an alcohol has an anti-arteriosclerotic activity by inhibiting the oxidation of low density lipoprotein (hereinafter LDL) A health functional food for prevention and improvement of health.

Hereinafter, the present invention will be described in detail with reference to examples. However, these examples are intended to further illustrate the present invention, and the scope of the present invention is not limited to these examples.

[ Example ]

Preparation Example  One : The ( Abutilon theophrasti Medicus ) Preparation of leaf extract

Abutilon collected from Chungbuk theophrasti Medicus ) leaves were lyophilized and dried at -80 ° C, and pulverized and pulverized powdery leaf samples were prepared. Then, the leaf sample was extracted with ethanol at 50 ° C. using an accelerator solvent extraction apparatus to prepare an edible leaf extract. Ethanol contained in the edible leaf extract was removed using a vacuum concentrator.

Experimental Example  One : The  Cell viability inhibition experiment and antioxidant activity assay of leaf extracts.

Experimental Example  1-1. The  Inhibition of Cell Survival by Leaf Extract.

In order to confirm the cell survival rate of the leafy leaf extract of the present invention, Raw 264.7 cells, a macrophage of mice, were cultured from an American type culture collection. The Raw 264.7 cells were plated in a 24-well plate at a density of 7 × 10 ⁴ / well per well and cultured for 16 hours. Then, the cultured cells were treated with the edible leaf extract prepared in Preparation Example 1 at a concentration of 5, 10, 20, 40 uM / ml and cultured for 24 hours. Then, 3- (4,5-dimethylcyazol- (3,5-dimethylthiazol-2-yl) -2,5-diphenyltetra zolium bromide (MTT). Afterwards, the formazan produced by living cells was measured for absorbance at 540 nm in a microplate reader. Statistical analysis was performed using one way ANOVA by Newman-Keuls Multiple comparison test of prism 5 (Prism 5 for windows of Graphic Pad, California, USA) Respectively.

As a result of the above statistical analysis, it was confirmed that even when the concentration of the leaf blade extract was increased, it was not cytotoxic (Fig. 1).

Experimental Example  1-2. The  Of leaf extract Peroxynitrite  Production inhibition effect measurement experiment.

In order to confirm the inhibitory effect of peroxynitrite (ONOO-) production, which is a strong oxidative stress, as a compound of active nitrogen and active oxygen in the leaf blade extract of the present invention, 3-morpholinoside nonimine hydrochloride (3 1 μl / ml, 2 μl / ml, or 4 μl / ml of mouse leaf extract solution (0.5 μl / ml) and 50 μl of sodium phosphate buffer (pH 7.4) supplemented with 50 μM of morpholinosyd nonimine hydrochloride (SIN- Ml was mixed to prepare a mixed solution. The sodium phosphate buffer contains 5 mM KCl, 100 [mu] M diethylenetriaminepentaacetic acid and 10 [mu] M DHR 123 (dihydrorhodamine 1,2,3). The mixture was then subjected to fluorescence measurement in a fluorescence microplate reader at 485 nm / 530 nm (excitation wavelength / emission wavelength).

Statistical analysis was performed using one way ANOVA by Newman-Keuls Multiple comparison test of prism 5 (Prism 5 for windows of Graphic Pad, California, USA) Respectively.

As a result of the above statistical analysis, it was confirmed that the leafy leaf extract inhibited the peroxynitrite produced by 50 uM SIN-1 in a concentration-dependent manner with a small amount of 0.5-4 μg / ml (see FIG. 2).

The fluorescence of peroxynitrite inhibition effect induced by SIN-1 treatment of macrophage with macrophyllite was measured by microplate reader. As a result, SIN-2 and SIN-1 It was confirmed that the effect of peroxynitrite inhibition was significantly increased in the cells treated with the leaf leaf extract (Fig. 2). Therefore, the leafy leaf extract has antioxidant activity due to the effect of inhibiting peroxynitrite.

Experimental Example  1-3. The  Experiments on the inhibitory effect of leaf extract on nitrogen monoxide formation.

Raw 264.7 cells, such as the Raw 264.7 cells used in Experimental Example 1-1, were added to a 48-well plate at a concentration of 50 x 10 ⁴ / well in order to confirm the inhibitory effect on the production of nitrogen monoxide by the diabetic leaf extract of the present invention And cultured for 16 hours. Subsequently, 100 ng / ml of lipopolysaccharide (Sigma) was added to the cultured cells, and 5 μM / mL, 10 μM / mL, 20 μM / mL or 40 μM / Ml and cultured for 18 hours. The amount of nitrite secreted in the supernatant was then measured with a microplate reader at 550 nm using a griess reagent system.

As a result of the measurement, the leaf blade extract inhibited the production of nitrogen monoxide (FIG. 3) in a concentration-dependent manner, and inhibited 50% of nitrogen monoxide produced by LPS at 6.9 / / And inhibited the formation of nitrogen monoxide, which was more than 11.5 占 퐂 / ml (Table 1).

division NO IC ₅₀ (required for 50% inhibition of nitrogen monoxide) Diabetic leaf extract 6.9 占 .8 占 퐂 / ml Penicillamine 11.5 占 .1 占 퐂 / ml

Thus, the leafy leaf extract inhibits the production of nitrogen monoxide in a concentration-dependent manner, and a lesser amount of penicillamine, which is a positive control substance, is used to inhibit the nitrogen monoxide produced by LPS, (Fig. 3).

Experimental Example  2 : The  Inhibition of In Vitro Inflammatory Protein Expression in Leaf Extracts.

The leaf blade extract of the present invention may be used as an anti-inflammatory protein, such as cyclooxygenase-2 (COX-2), nitric oxide synthase (iNOS) or nuclear factor kappa B -kB), Raw 264.7 cells such as Raw 264.7 cells used in Experimental Example 1-1 were divided into 6 × 10 ⁵ cells in a 100 mm plastic flask and cultured for 16 hours. Then, 5 μg / ml, 10 μg / ml, 20 μg / ml or 40 μg / ml of the leafy leaf extract was added and treated with 1 μg / ml of lipopolysaccharide (LPS) Time. After culturing, Raw 264.7 cells were washed with phosphate buffered saline (PBS), and the cells were treated with 100 μl of cytoplasmolysis solution, left on ice for 15 minutes, and centrifuged to separate the supernatant. The cytoplasmic lysate was dissolved in 10 mM Tris (pH 8.0), 1.5 mM MgCl ₂ , 1 mM DTT (dithiothreitol), 40 mol% (NP-4; Nonylpenol 40 mole Ethoxylates) and protease inhibitors .

The separated supernatant is a cytoplasmic protein for confirming COX-2, iNOS and NF-kB expression. The separated supernatant proteins were electrophoresed on a 5% SDS (sodium dodecyl sulfate) -acrylamide gel and then transferred to a nitrocellulose membrane. Thereafter, the membrane was incubated with TBST containing 5% skim milk for 30 minutes at room temperature to inhibit non-specific binding of the antibody. The TBST is a mixture of 50 mM Tris (pH 7.5), 150 mM NaCl, 0.1% Tween-20 (Polyoxyethylene (20) sorbitan monolaurate).

Then, the cells incubated at room temperature were reacted with specific antibodies of COX-2, iNOS and NF-kB at 4 ° C for 24 hours. The expression levels of COX-2, iNOS and NF-kB were secondary antibodies with horseradish peroxidase (HRP) attached at 25 ° C for 30 min. Respectively.

As a result of the X-ray film analysis, it was confirmed that Raw 264.7 cells of mouse leaf extract inhibited the expression of COX-2, iNOS and NF-kB caused by treatment with 1 μg / ml of LPS in a concentration-dependent manner Reference).

Inhibition of iNOS, COX-2 and NF-kB expression by inflammatory cytokines induced by lipopolysaccharide (LPS) treatment of macrophyllous leaf extracts resulted in the inhibition of LPS The expression levels of iNOS, COX-2 and NF-kB were significantly reduced in the cells treated with the leafy leaf extract (FIG. 4).

Experimental Example  3: The  Low density lipoprotein oxidation of leaf extract Low performance  Analysis experiment.

In order to confirm the oxidation-inhibitory effect of the low-density lipoprotein (LDL; low-density lipoprotein), which is a factor promoting arteriosclerosis of the present invention, 50 to 100 μg of protein was dissolved in human blood Prepare isolated LDL. Then, 0.02 ml of a leaf leaf extract at a concentration of 20 μg / ml (= ppm) or 100 μg / ml (= ppm), 0.115 ml of 10 mM phosphate buffer (PBS) and 0.04 ml of 0.25 mM CuSO _{4 were} added to the prepared LDL To prepare a reaction solution, and the reaction solution was allowed to react at 37 ° C for 3 hours. Then, 1 ml of 20% TCA (Trichloroacetic acid) solution was added to the reaction solution, and the reaction was stopped. 1 ml of 0.67% TBA (Thiobarbituric acid) solution dissolved in 0.05 N NaOH was added and the mixture was heated at 95 ° C for 15 minutes and then cooled for 15 minutes .

The cooled reaction solution was centrifuged at 3,000 rpm for 15 minutes, and the amount of malondialdehyde (MDA) contained in the separated supernatant was measured at 540 nm for statistical analysis.

The statistical analysis was performed using statistical analysis systems (SAS) program at a level of p <0.05 by one way ANOVA and Duncan's multiple range test.

As a result of the above statistical analysis, the leaf blade extract showed higher LDL oxidation inhibitory effect than ascorbic acid (L-ascorbic acid) at the concentration of 20 ppm. In addition, at 100 ppm concentration, the leaf blade extract showed an LDL oxidation inhibition rate of 20.7 ± 0.7%, which was similar to that of 23.9 ± 0.9% of ascorbic acid, a positive control substance (see FIG. 5 ).

The LDL oxidation inhibitory effect of 20 leaf or 20 ppm LDL extracts from human blood was investigated. The results showed that LDL oxidation inhibition effect in cells treated with mouse leaf extract and ascorbic acid (Fig. 5). As shown in Fig.

As can be seen from the above experimental examples, the leafy leaf extract of the present invention inhibits the oxidation of low density lipoprotein (LDL) and has atherosclerotic activity, and cyclooxygenase-2 (COX-2 ), Nitric oxide synthase (iNOS), or nuclear factor kappa B (NF-kB), and thus has anti-inflammatory activity. Also, it can be confirmed that the leaf blade extract inhibits the production of peroxynitrite (ONOO ^- ) or nitric oxide (NO ^- ) and thus has an antioxidative activity.

[ Manufacturing example ]

Manufacturing example  1: Preparation of pharmaceutical composition

The formulation of a pharmaceutical composition comprising the leafy leaf extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically described.

Manufacturing example  1-1. Sanje  Produce

Diabetic leaf extract 2 g

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

Manufacturing example  1-2. Manufacture of tablets

100 mg of leafy leaf extract

Corn starch 100 mg

100 mg of milk

Magnesium stearate 2 mg

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

Manufacturing example  1-3. Preparation of capsules

100 mg of leafy leaf extract

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

Manufacturing example  1-4. Manufacture of rings

1 gram of leafy leaf extract

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

Manufacturing example  1-5. Manufacture of granules

150 mg of leafy leaf extract

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

Manufacturing example  2: Preparation of health functional foods.

Manufacturing example  2-1. Manufacture of Health Functional Foods

3000 mg of leafy leaf extract

70 [mu] g of vitamin A acetate

Vitamin E 1.0 mg

0.13 mg vitamin B1

0.15 mg of vitamin B2

10 mg vitamin C

Biotin 10 μg

50 ㎍ of folic acid

Calcium pantothenate 0.5 mg

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

15 mg of potassium phosphate monobasic

Potassium citrate 90 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixture is relatively mixed with a suitable preparation for health food, the compounding ratio may be arbitrarily changed, and the above ingredients may be mixed according to a conventional method for producing healthy food , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Manufacturing example  2-2. Manufacture of health drinks

Algae leaf extract 3 g

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 1 g

Purified water was added to a total of 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated for about 1 hour at 85 DEG C with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, Of health functional beverage composition.

Although the composition ratio is relatively mixed with the ingredients suitable for the favorite beverage as a preferable preparation example, the blending ratio may be arbitrarily modified according to the local or national preference such as the demand class, demand country, use purpose, and the like.

Claims (7)

( Abutilon theophrasti Medicus ) leaf extract as an active ingredient,
Prevention and treatment of inflammatory diseases which inhibit the expression of inflammatory proteins in the body containing at least one of the group consisting of cyclooxygenase-2 (COX-2) and nitric oxide synthase (iNOS) A pharmaceutical composition. The method according to claim 1, wherein the inflammatory disease comprises at least one selected from the group consisting of dermatitis, sore throat, tonsillitis, fibromyalgia, rheumatoid arthritis, shoulder joint inflammation, myositis, hepatitis, cystitis and nephritis. A pharmaceutical composition for therapeutic use. The pharmaceutical composition according to claim 1, wherein the leaf blade extract is an extract of at least _one of C ₁ -C ₄ alcohols. delete (Abutilon theophrasti Medicus) leaf extract as an active ingredient,
Prevention and improvement of inflammatory diseases that inhibit the expression of inflammatory proteins in the body containing at least one of the group consisting of cyclooxygenase-2 (COX-2) and nitric oxide synthase (iNOS) Health functional food for. Claim 5 wherein said Abutilon theophrasti leaf extract is C ₁ ~ C ₄ health functional food for the prevention and improvement of inflammatory diseases characterized in that the extract is extracted with one or more of the alcohol. 6. The method according to claim 5, wherein the inflammatory disease comprises at least one of the group consisting of dermatitis, sore throat, tonsillitis, fibromyalgia, rheumatoid arthritis, shoulder joint inflammation, myositis, hepatitis, cystitis and nephritis. Health functional food for improvement.

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