KR20160001280A - Composition Comprising Extracts of Boehmeria tricuspis, Angelica decursiva and Arctium lappa Having Anti-allergy and Anti-inflammation - Google Patents

Abstract

Disclosed is a composition comprising a complex extract of Boehmeria tricuspis, Angelica decursiva, and Arctium lappa for anti-allergic and anti-inflammatory activities. The complex extract of Boehmeria tricuspis, Angelica decursiva, and Arctium lappa, extracted with water or ethanol, and alcohol of the present invention can inhibit expression of iNOS and COX-1/2 which are inflammation-mediated factors, and can inhibit secretion of β-hexosaminidase which is an indicator of an allergic reaction. Accordingly, the effects of a complex function can be maximized by combination of three types of effective plants and a suitable extraction method.

Description

TECHNICAL FIELD The present invention relates to an anti-skin allergic and anti-inflammatory composition containing a complex extract of turtle tail, body herb and burdock.

The present invention relates to an anti-skin allergic and anti-inflammatory composition containing a complex extract of turtle tail, body herb and burdock.

Atopic dermatitis accompanied by itching is common in early infancy and often accompanies allergic diseases such as atopic asthma and allergic rhinitis among patients and family members. The main symptom of atopic dermatitis is itch, and various symptoms such as dry skin, keratosis of pores, eczema, and white skin necrosis may occur. Patients with atopic dermatitis have elevated serum IgE and are often positive when tested for allergic skin reactions and their symptoms are aggravated by environmental or emotional factors.

More than 50% of patients develop within 3 months to 1 year after birth and 30% develop between 1 and 5 years. That is, most cases are those that develop before 5 years of age. 80% of patients may develop asthma or rhinitis during boyhood, and skin symptoms often improve when respiratory allergies occur. Atopic dermatitis is generally more severe and persistent in infants and tends to improve with age. In other words, between two and three years, about 80% of the symptoms improve but sometimes it continues to be adults. The cause of atopic dermatitis has not yet been clarified yet, but about 30% of pediatric patients are known as food allergies.

To treat atopic dermatitis, moisturizers are used on dry skin to keep the skin from drying, and corticosteroids to treat dermatitis, and appropriate antihistamines to treat pruritus or sleep disorders caused by it. The use of adrenocortical hormones is limited due to the high risk of adverse effects due to endocrine disruption in the human body, and natural products that inhibit histamine secretion can be used to improve atopic skin diseases. Atopic skin diseases are accompanied by bacterial infections and inflammation of scars caused by scratching along with itching. The cause of the inflammation reaction is physiological, chemical action or bacterial infection. It is a defense mechanism of the body. It removes foreign substances and restores the damaged area. Once the stimulus is applied, histamine, serotonine , bradykinin, prostaglandins, hydroxyeicosatetraenoic acid (HETE), and leukotriene are released, resulting in increased vascular permeability and inflammation.

Monocytes are activated by bacterial infections and transformed into macrophages with excellent phagocytosis. Macrophages induce inflammation by secretion of NO, prostaglandin E2, and proinflammatory cytokine, and are important regulatory cells in both inflammation and immune responses. In order to function like this, it must be activated. LPS (lipopolysaccharide), one of the cell wall components of Gram-negative bacteria and well known as endotoxin, is the most well-known external factor involved in macrophage activation. Pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β (interleukin-1 beta) have been observed in monocytes and macrophages such as RAW 264.7 cells. inflammatory cytokines secreted by the inflamed area is increased.

When LPS stimulates macrophages, nitric oxide (NO) is produced in the process of converting L-arginine into L-citrulline by an enzyme called iNOS (inducible nitric oxide synthase), resulting in the generation of NO from macrophages. In mammals, NO is synthesized by three kinds of nitric oxide synthase (NOS): nNOS (neuronal NOS), eNOS (endothelial NOS) and iNOS (inducible NOS). Among these, NO produced by nNOS and eNOS is produced for normal biological function, and concentration in tissues is kept low to a certain level.

However, NO produced by iNOS is overexpressed and exhibits deleterious effects on living organisms such as pathological vasodilation, cytotoxicity, and tissue damage. Prostaglandin E2 (PGE2), an inflammatory factor, is produced by the action of an enzyme called COXs (cyclooxygenaes), which is released by an enzyme called phospholipase A2, And induce an inflammatory reaction. COX is classified as COX-1 and COX-2. COX-1 acts on normal biological functions such as platelet formation, gastric wall protection and maintenance of renal function in the body, and COX-2 synthesizes PGE2 as an inflammatory mediator . PGE2 is known to be deeply involved in the development of cancer, such as promoting inflammation (pain, fever, etc.), immune response, and angiogenesis. All nonsteroidal antiinflammatory drugs (NSAIDs), including selective COX-2 inhibitors, exhibit anti-inflammatory, antipyretic and analgesic effects. However, long-term use of these drugs has serious side effects. For example, secondary anemia due to gastrointestinal peptic ulcer bleeding, platelet function suppression, inhibition of induction of labor, side effects to the kidney, liver damage, hypersensitivity. In order to overcome the side effects of these drugs, natural products such as plants are more preferable as a safer material. In this regard, researches are being actively carried out to find and use the anti-inflammatory materials in natural products.

1. Korean Registered Patent No. 10-0556524 (Feb. 23, 2006) 2. Korean Patent No. 10-1219520 (2013.01.02)

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made in view of the above problems, and it is an object of the present invention to provide an anti-skin allergic and anti-inflammatory composition containing a complex extract of turtle tail, body herb and burdock.

In order to achieve the above object, an embodiment of the present invention is characterized by a composition for anti-skin allergy and anti-inflammation containing a complex extract of turtle tail, body herb and burdock.

Another embodiment of the present invention is a composition for anti-skin allergy and anti-inflammation comprising a fraction obtained by fractionation of ethyl acetate from a complex extract of turtle tail, body herb and burdock.

The method for preparing anti-allergy and anti-inflammatory composition according to the present invention comprises shredding a turtle tail, a body herb and a burdock complex extract to finely pulverize a dried plant to obtain a powder; Extracting the mixed composite with an alcohol solvent such as water or alcohol; And fractionating the extracted complex extract with ethyl acetate to extract fractions.

The complex extraction step is carried out at 20 to 100 ° C with an alcohol solvent such as water or alcohol at a volume of 3 to 20 times the dry weight of the composite, filtered under reduced pressure, and the filtrate is concentrated under reduced pressure at 20 to 100 ° C in a vacuum rotary condenser The method comprising the steps of:

The step of extracting the fraction includes dissolving the extracted complex extract using water, and further fractionating the extract into an ethyl acetate layer and concentrating the extract under reduced pressure.

Indicators of the alcohol, alcohol or H ₂ O extracts of the composites of the turtle tail body herbs and burdock of the present invention not only to inhibit the iNOS and COX1 / 2 generation induced by LPS in RAW 264.7 cells, histamine release inhibition (indicator) It is possible to use the composition as a composition for the prevention and treatment of skin allergies and inflammatory diseases by effectively inhibiting beta-hexasaminidase.

FIG. 1 is a diagram showing a large-scale extraction process for producing a complex extract of turtle tail, body herb and burdock.
FIG. 2 is a graph showing the effect of the ethyl acetate complex extract (EP) of turtle tail, body herb and burdock to inhibit the expression of iNOS as an inflammatory mediator in RAW 264.7 cells.
Figure 3 shows the inhibitory effect of COX (cyclooxygenase 1/2), an important factor in the inflammation mediator of the ethyl acetate complex extract (EP) of turtle tail, body herb and burdock in RAW 264.7 cells.
FIG. 4 shows the effect of suppressing the secretion of β-hexosaminidase, an index factor of allergic reaction of ethyl acetate complex extract (EP) of turtle tail, body herb and burdock in RBL-2H3 cells.
FIG. 5 shows the results of cytotoxicity tests (MTT assay) of ethyl acetate complex extract (EP) of turtle tail, body herb and burdock in RAW 264.7 cells.

Turtle tail, body herb and burdock are native plants in Korea.

Turtle tail (Scientific name: Boehmeria tricuspis) is a perennial plant with nettle, and Korea is distributed in Jeju, Jeonnam, Gyeongnam, Gangwon, Gyeonggi, Pyeongnam, Hamnam, Japan, and China. It grows in the forest edge or a little shady. Stems are united, reaching 1m in height, squashed, stood straight, branches are split, red with petiole. The leaves are opposite and egg-shaped, the ends are divided into 3 branches, and the cracks in the middle are 25cm long and become like a turtle tail. 3 Macs are distinctive, have large saw teeth, and have hairs on their back veins. Flowers are bloomed in August and August, and light green flowers run on the axilla with watery flowers [穗 角花序], with male flower on the bottom of the stem and female flower on the top. Male flower has 45 calyx and 45 stamens. Several female flowers gather in small ball shape and are wrapped in a tubular calyx, with one style. Fruits are ovate (果果) in the form of inverted ovary or several gathered round. Stems are for fiber, young leaves for food.

Body herb (Scientific name: Angelica decursiva) is a perennial herb that grows near mountains and wetlands. Roots are short and roots are thick. The stem is straight and has a dense vertical line, the branch is divided at the upper part and the height is 80 ~ 150cm. Leaves are alternate phyllotaxis, 35 small leaves. Small leaves are deeply or fully split into 35 pieces, and the lower part flows downward into a wing shape with sawtooth on the edge. Leaves from the roots and leaves on the bottom of the stem have long petiole, and the leaves on the upper part of the stem cover the stem with the lower part of the petiole becoming the sheath. The flower blooms in October with a deep purple color and runs in a large double inflorescence. The small peduncle with umbrella shape is 10 ~ 20, 20 ~ 30 flowers each. There are twelve guns, wide guns, and 57 small guns. The fruit is a division (fruit divided from division) and it is a flat oval shape. Young seeds eat as herbs. It uses roots as herbal medicine called "Hu Hu" in the oriental medicine, and it is effective for cold coughing asthma by acting as a fever gland. It is distributed in Korea, Japan, China and others.

Burdock (Scientific name: Arctium lappa) is a biennial plant of Asteraceae which grows 50 to 150 cm high. Straight roots grow 30 ~ 60cm and stem comes out from the end. The leaves on the roots come out in rows and the petiole is long. It is staggered and heart-shaped in the trunk. The surface is dark green, but the back side has white fluffy dense, with serrated teeth at the edge. Flowers bloom from July to August, with purplish purple spots. The gun is round, the bract is needle-like, and the tip is like a hook. The flowers are tubular, the seeds are black and the tubers are brown. The fruit is achene and ripens in September. It is sturdy, hardly sick, very strong in the cold, and does not cover much of the soil. Breeding is done by seeding or giving up. It is a naturalized plant of European origin. The varieties have long roots, thick agar, good quality meat, and short roots. The recipes are made with pickles or boiled and eaten as side dishes. The roots contain inulin and some palmitic acid. In Europe, it is used as a diuretic and sweating agent. Seeds are used as diuretics when there is a swelling, and as an antidote to sore throats and poisonous insects. It grows much in Japan and is wild in Europe, Siberia and northeastern China.

In these three plants, turtle tail, body herb and burdock are shredded, dried and then extracted with ethanol or water (H ₂ O) and concentrated under reduced pressure.

The extract is separated into ethyl acetate layer and concentrated under reduced pressure to obtain a combined extract composition for use in improving skin allergy and inflammation.

The present invention comprises extracting from a complex extract of turtle tail, body herb and burdock with water or an organic solvent, especially alcohol and alcohol, and fractionating with ethyl acetate.

The method for preparing the complex extract according to the present invention is as follows.

1. Turtles tail, body herbs and burdock shrubs dried plants to pulverize.

2. Extract the mixed composite with an alcohol solvent such as water or alcohol.

At this time, a volume of about 3 to 20 times, preferably about 3 to 10 times, the volume of the dry weight of the composite is dissolved in an alcohol solvent such as water, alcohol, preferably water or alcohol at room temperature or 20 to 100 占 폚, Is subjected to the extraction method such as hot water extraction, cold extraction, warming or reflux cooling extraction or ultrasonic extraction for 1 to 5 times at an extraction temperature of 20 to 70 캜 for about 0.5 to 2 days, preferably 1 to 1 day, Preferably three consecutive times, is obtained by filtration under reduced pressure, and the filtrate is concentrated in a vacuum rotary condenser at a temperature of 20 to 100 ° C, preferably 40 to 70 ° C under reduced pressure to give a water, an alcohol or a mixed solvent thereof An extract can be obtained.

3. Extract the combined extracts with ethyl acetate to extract fractions.

The extracted complex extract is dissolved with water and then further fractionated into an ethyl acetate layer and concentrated under reduced pressure to prepare an ethyl acetate fraction.

The present invention relates to an extract of fractions of a turtle tail, a body herb and a burdock, fractions fractionated therefrom, and a preparation method thereof, wherein the complex extract or the ethyl acetate fraction has excellent anti-inflammatory effects on the antiallergic and inflammatory reaction of the active ingredient .

A pharmacologically acceptable emulsifier, diluent excipient or the like may be added to the plant extract composition for anti-skin allergic or anti-inflammatory activity, or a base material for the production of cosmetics may be added.

The plant extract composition for improving skin allergy and inflammation of the present invention may be in a form selected from a solution, a suspension, an emulsion, a gel, a cream, a lotion, a powder, a soap, a cleansing, a body wash, a cleansing and a spray, . ≪ / RTI >

The composite extract composition of turtle tail, body herb and burdock of the present invention has an advantage of excellent skin allergy and inflammation improving effect. This is for the purpose of illustrating the present invention, and the present invention is not limited by these examples.

<Examples> Production of a complex extract of turtle tail, body herb and burdock

The ethyl acetate complex extract (EP) of the turtle tail, the body herb and the burdock used in the present invention was prepared by shredding shrubby dried plants and mixing 1 kg in a weight ratio of 1: 1: 1, adding 3 L of methane (ethane) After repeating the extraction three times at intervals, the filtrate was filtered through a filter paper (Wattsman, USA), and the filtrate was subjected to removal of the solvent at 45 ° C with a vacuum rotary condenser, and 120 to 220 g of SD was obtained as an extracted residue. After dissolving in water (H ₂ O), the residue was fractionated into an ethyl acetate layer and concentrated under reduced pressure to give an ethyl acetate fraction (FIG. 1).

Experimental Example 1 Effect of Ethyl Acetate Composition Extract (EP) on Turtle Tail, Body Herb and Burdock for iNOS Production

RAW 264.7 cells (10 × 10 ⁴ cells / well) were inoculated into a 24-well plate using DMEM medium, and after 12 hours, fresh medium containing the sample and LPS (1 μg / ml) were simultaneously treated and cultured for 24 hours. The amount of NO produced was measured using the Griess reagent in the form of NO ^{2 -} present in the cell culture. 100 μl of cell culture supernatant and 100 μl of Griess reagent (1% sulfanilamide, 0.1% naphtylethylenediamine in 2.5% phosphoric acid) were mixed and reacted on 24-well plates for 10 minutes. Absorbance was measured at 540 nm using an ELISA reader. The results are shown in Fig. 2 and Table 1 below. As can be seen in FIG. 2, it was confirmed that EP effectively reduced the production of iNOS, an inflammatory mediator, in RAW 264.7 cells induced by LPS in a concentration-dependent manner.

Experimental Example 2 Evaluation of cyclo-oxygenase-1/2 inhibition by EP

The screening process for the screening of COX-1/2 inhibitors against EP was performed according to the method of Kaiman in the following manner. Briefly, COX-1/2 (Cayman, Cayman) was incubated in an incubate with the diluted extract at a ratio of 1: 500 for 15 minutes. After incubation, Quantablu (Pierce) substrate was added and allowed to mature for 45 minutes at 25 ° C. Maturation was followed by reading the luminescence with a Wallac Victor plate reader, The results are shown in FIG. 3 and Table 1. As can be seen from FIG. 3, it can be confirmed that EP effectively reduces the production of COX in a concentration-dependent manner in RAW 264.7 cells induced by LPS.

<Experimental Example 3> Anti-allergic effect of ethyl acetate complex extract (EP) of turtle tail, body herb and burdock

The secretion of β-hexosaminidase was measured in order to examine the inhibitory effect on degranulation, which is an index of allergic response. RBL-2H3 cells were suspended in DMEM containing 10% FBS, and the cells were distributed to a 48-well plate (Corning, NY, USA) at a density of 5 × 10 ⁵ cells / ml. Subsequently, the cells were sensitized with anti-DNP IgE (30 ng / ml) and incubated for 24 hours at 37 ° C in a 5% CO ₂ incubator. Cells of each well were washed twice with buffer (119 mM NaCl, 5 mM KCl, 0.4 mM MgCl 2, 25 mM PIPES, 40 mM NaOH, pH 7.2) and then 5.6 mM glucose, 1 mM CaCl ₂ and 0.1% BSA was added and the cells were incubated with DNP-HSA (10 μg / ml) for 1 h in concentrations of 0, 2, 10 and 50 μ / Lt; / RTI &gt; 40 μl of the supernatant was transferred to a 96-well plate, and 80 μl of substrate buffer (1 mM p-nitrophenyl-N-acetyl-bD-glucosaminide, 0.05 mM sodium citrate, pH 4.5) was added and incubated at 37 ° C for 30 minutes. 200 μl of stop solution was added to each well to terminate the reaction. Absorbance was measured at 405 nm using a microplate reader (Molecular Devices Co. Ltd., USA). The inhibition (%) was calculated from the absorbance values of the EP treatment group and the control group according to the following equation, and the results are shown in FIG. 4 and Table 1.

Inhibition (%) = (1-T / C) x 100

C: Cell (+), DNP-HSA (+), test sample (0)

T: Cell (+), DNP-HSA (+), test sample (+)

<Experimental Example 4> Cytotoxic effect of ethyl acetate complex extract (EP) of turtle tail, body herb and burdock

The cytotoxicity of RA-264.7 cells, which were AD-macrophages obtained in the above-mentioned Example, was measured in order to confirm anti-inflammatory activity in an appropriate concentration by treating LPS (1 ㎍ / ml). RAW 264.7 cells (10 × 10 ⁴ cells / well) were treated with test drug (EP) in DMEM medium and cultured for 24 hours. 500 μl of culture medium and 5 μl of MTT solution were added to a 24-well plate with a new medium, followed by further reaction for 4 hours. After removing the medium, formazan was dissolved in 200 DMSO and the absorbance was measured at 540 nm using a microplate reader. The measurement value is an average value of three repeated experiments, and the results are shown in Fig. 5 below. As shown in FIG. 5, the cytotoxicity of EP to RAW 264.7 cells was not significantly elevated to a concentration of 100 μg / ml.

sample ^* IC ₅₀  (g / mL) iNOS COX1 / 2 β-hexosaminidase MeOH extract 21.3 25.4 35.3 The EtOAc fractions (EP) 7.6 9.1 12.8

* IC ₅₀ : 50% inhibitory concentration

Claims (5)

A composition for anti-allergic and anti-inflammatory skin comprising a complex extract of turtle tail, body herb and burdock. A composition for anti-allergic skin and anti-inflammatory, which comprises a fraction obtained by fractionating from a complex extract of turtle tail, body herb and burdock with ethyl acetate. Shredding the turtle tail, the body herb and the burdock complex extract to finely pulverize the dried plant;
Extracting the mixed composite with an alcohol solvent such as water or alcohol; And
And separating the extracted complex extract into an ethyl acetate layer to extract fractions. The method for producing anti-allergy and anti-inflammatory composition according to claim 1, 4. The method of claim 3,
Extracting the solution at 20 to 100 DEG C with an alcohol solvent such as water or alcohol having a volume of 3 to 20 times the dry weight of the complex, filtering the mixture under reduced pressure, and concentrating the filtrate under reduced pressure at 20 to 100 DEG C using a vacuum rotary condenser Wherein the anti-allergic agent is an anti-allergic agent. The method according to claim 3,
Dissolving the extracted complex extract with water, and then fractionating the extract into an ethyl acetate layer and concentrating the extract under reduced pressure.

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