KR20180090751A - Composition for preventing or treating inflammatory disease comprising Angelica gigas extract and lactic acid bacteria - Google Patents

Abstract

The present invention relates to a composition for preventing or treating inflammatory diseases comprising an Angelica gigas extract and lactic acid bacteria. The composition of the present invention includes a complex of the Angelica gigas extract and lactic acid bacteria, thereby not causing side effects in the human body; and having the excellent treatment effect of colitis including the reduction of body weight, decrease of colon length, and increase of disease activity in a colitis animal model compared to a single-dose group. Additionally, the composition of the present invention has therapeutic effects on atopic dermatitis and treats pruritus, and thereby can be usefully used in the field of pharmaceuticals and health functional foods for the treatment of inflammatory diseases.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for preventing or treating an inflammatory disease, including Angelica gigantosa extract and lactic acid bacteria,

The present invention relates to a composition for preventing or treating an inflammatory disease, including Angelica gigantosa extract and lactic acid bacteria.

Inflammatory disease refers to the pathological state of abscesses that are formed by the invasion of bacteria. Inflammation is a normal and protective in vivo defense manifestation that is localized to physical trauma, harmful chemicals, microbial infections, or tissue damage caused by stimuli in vivo metabolites. These inflammations are triggered by various chemical mediators produced from damaged tissues and migrating cells, and these chemical mediators are known to vary according to the type of inflammation process. In normal cases, the organism neutralizes or eliminates the cause of inflammation through the inflammation reaction, regenerates the upper tissue and regenerates the normal structure and function, but if not, the disease state such as chronic inflammation also proceeds. In addition, when the inflammation is improperly induced by an autoimmune reaction such as a harmless substance such as pollen, asthma or rheumatoid arthritis, the defense reaction itself rather damages the tissue, and therefore, an inflammatory disease prevention or treatment agent is required. Inflammatory reactions can be observed in almost all clinical diseases. Some of these inflammatory diseases are bacterial diseases that can be cured by antibiotic treatment, but most of them are due to tissue damage due to autoimmune reaction, so there is no specific treatment It is known as an incurable disease.

One example of an inflammatory disease is atopic disease. In atopy-related diseases, atopy has the meaning of 'atopos', the Greek word 'unusual', 'strange', 'abnormal reaction'. Literally, a variety of causes are complicated, muddled, loosened and recurred. Atopic diseases include asthma, conjunctivitis, and dermatitis. These diseases may occur singly or in various diseases. In general, immunosuppressive inflammatory diseases including atopic dermatitis are typical examples. Immune hypersensitivity reactions, including atopic dermatitis Inflammatory diseases are one of the most common skin diseases in infants and children and begin at a rate of 45% at 6 months, 60% before 12 months and at least 85% before 5 years of age. Although it is usually known to be a short-term illness at a young age, 50% of the patients disappear within two weeks, but 25% continue until adolescence and the remaining 25% continue without disappearance (Thomas Bieber, Atopic Dermatitis, The New England Journal of Medicine, 2008, Vol. 358, ISSN 0028-4793, 1483-1494). The cause of atopic dermatitis has yet to be clarified. However, the level of serum IgE antibody due to the immune response to the antigenic material introduced through the defective skin layer, the level of genetic family history, (Park, Chan-Ik, et al., The effect of Huang Ryun Ha-doktang on NC / Nga Mice-induced atopic dermatitis, Kor. J. Herbology, 2008, 23 (2): 59-65). Although the cause of the immune-sensitized inflammatory disease including atopic dermatitis, which is known to date, is not known precisely, it is generally believed that genetic and immunological factors are involved, and other environmental and psychological factors act as deteriorating factors It is a universal viewpoint. The main symptom of atopic dermatitis is itch. When it gets scratched, it develops as an eczematous lesion, and as the lesion progresses, a series of vicious cycle of repeated pruritus is repeated. In addition, immunosuppressive inflammatory diseases including atopic dermatitis are characterized as refractory allergic diseases. Total immunoglobulin E (IgE) values are increased in more than 80% of patients with immunosuppressive inflammatory diseases including atopic dermatitis, and allergic rhinitis Or asthma, it is known that the level of immunoglobulin E is high (Thomas Bieber, Atopic Dermatitis, The New England Journal of Medicine, 2008, Vol. 358, ISSN 0028-4793, 1483-1494). Currently, dexamethasone, which is known to be a steroid agent, is the most widely used treatment for immunosuppressive inflammatory diseases including atopic dermatitis. However, it is effective for short-term treatment, but it is not stable and effective in long- (Such as Sinillan, German chamomile, lavender, sandalwood mixed oil, atopic dermatitis model NC / Nga mice), and the like are reported to be caused by side effects such as skin thinning, skin atrophy, Effects, 2009, Daejeon University Theses, p. 13). As such, synthetic drugs have problems in long-term use, and since they are not well treated with one active ingredient, there is a growing interest in extracts derived from natural materials having various active ingredients and functions in recent years.

Another example of an inflammatory disease is ulcerative colitis. In Western countries, the incidence of ulcerative colitis is 6 to 8 per 100 000 population, with a prevalence of 70 to 150, with similar male and female ratios. It occurs frequently in young people between 15 and 35 years of age, . In Korea, ulcerative colitis has been reported since the 1970s. The incidence and prevalence of ulcerative colitis has not been higher than that of Western countries. Recently, the incidence of inflammatory bowel disease has increased rapidly, , And the development of diagnostic technology has contributed to this. The cause of ulcerative colitis is not yet known, but environmental factors and genetic factors as well as excessive immune responses of our bodies to bacteria normally present in the intestines are considered to be important causes. Ulcerative colitis is a typical inflammatory bowel disease, which is caused by necrosis of the mucous membranes of the large intestine, inflammation or ulceration (mucosal layer of the inner wall of the oral cavity, esophagus, stomach, intestines, bladder, gallbladder, The defect is a chronic recurrent disease that has not yet been identified. Generally, it starts from the rectum adjacent to the anus and progressively progresses to the inside. It is characterized by continuous inflammation sites, with 50% of rectalitis, 25% of left colitis, and 25% of broad colitis.

Symptoms of ulcerative colitis include occasional symptoms of bleeding, and when the disease progresses, the stool becomes thinner and the number of bowel movements increases. If the bowel becomes more severe, it will go into the toilet dozens of times a day, and if it becomes worse, the ulcerative colitis There are no treatments available yet. However, drugs known to date are the most commonly used aminosalicylate sulfasalazine, mesalamine or corticosteroids, but severe immunosuppressants or antibiotics are used. These drugs, however, sometimes show different therapeutic effects in the same patient, and may cause inflammation in the liver, pancreas, and lungs from mild side effects such as nausea, heartburn, headache, dizziness, anemia and rash.

On the other hand, Angelica gigas (Nakai) is a perennial herbaceous perennial plant, and its flower is bloomed in September, purple, The young seeds are edible as herbs, the roots are called angelica, and they are used as medicines. It is distributed in Korea, China, Japan, etc., and it is called Angelica sinensis (Chinese origin), Angelica sinensis (Japanese mountain) and Angelica sinensis (Korean mountain). The medicinal properties of Angelica angustifolia are known to be weakness in the body, headache, dizziness, arthralgia, abdominal pain, constipation, menstrual irregularities and bruises.

Lactic acid bacteria are also referred to as lactic acid bacteria, and collectively refers to bacteria that ferment saccharides to obtain energy. Gram positive bacteria, lactic anaerobic or anaerobic. Lactobacillus and Streptococcus spp. Are known.

The present inventors have conducted studies to produce natural extracts having excellent anti-inflammatory activity. As a result, the present inventors have found that the complex of Angelica gigas Liquor and lactic acid bacteria has no harmful effect on the human body and has an excellent therapeutic effect in animal models of inflammatory diseases such as colitis and atopic dermatitis By confirming, the present invention has been completed.

The present invention provides a composition for preventing or treating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

The present invention also provides a food composition for preventing or ameliorating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

The present invention also provides a quasi-drug composition for preventing or ameliorating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

The present invention also provides a cosmetic composition for preventing or ameliorating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

Since the composition of the present invention includes a complex of angelica guinea pig extract and lactic acid bacterium, there is no side effect in the human body. Compared with the single administration group, the effect of treating colitis is reduced by reducing the body weight, decreasing the colonic length and increasing the disease activity, It is not only superior but also has improved itching and has a therapeutic effect on atopic dermatitis. Therefore, it can be usefully used in the field of pharmacy and health functional food for the treatment of inflammatory diseases.

Fig. 1 is a graph showing the results of analysis of weight change during an experiment period in an animal model of colitis. Fig.
FIG. 2 is a graph showing the results of measurement of colon length after completion of an experiment in an animal model of colitis.
3 is a graph showing the results of measurement of disease activity in an animal model of colitis.
4 is a graph showing the results of measurement of the number of scratches in an itchy animal model.
FIG. 5 is a graph showing the results of measurement of degree of skin damage in an atopic dermatitis animal model.
6 shows the results of analysis of the effect of the mixture of Angelica gigas Nakai extract and BP2 culture medium on IL-1β production induced by LPS.
FIG. 7 shows the results of analysis of the effect of the mixture of Angelica gigas Nakai extract and BP2 culture medium on the amount of TNF-α produced by LPS.

The present invention provides a composition for preventing or treating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

The composition includes a pharmaceutical composition, a health functional food composition, a quasi-drug composition or a cosmetic composition.

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

In the present invention, the Angelica giganta extract can be obtained by a conventional extraction method or a commercially available one can be purchased and used.

In the present invention, the "extract" means an extract obtained by extracting the Angelica gigas Nakai, a diluted solution or concentrate of the extract, a dried product obtained by drying the extract, a controlled preparation or a purified product of the extract, Itself and extracts of all formulations which can be formed using extracts.

Examples of the solvent that can be used to obtain the extract include water, alcohols having 1 to 4 carbon atoms, a mixed solvent thereof, and the like, but are not limited thereto.

Examples of the method for extracting the above extract include hot water extraction, cold extraction, reflux cooling, solvent extraction, steam distillation, ultrasonic extraction, elution, and compression. The extract may further be subjected to a conventional fractionation process, and may be purified using a conventional purification method.

In the present invention, the lactic acid bacteria can be used as an isolated strain or a variety of commercially available lactic acid bacteria without limitation. For example, the lactic acid bacteria include bacteria belonging to the genus Bacillus, Lactobacillus, Streptococcus, Bifidobacterium, Lactococcus, Pediococcus, Or Leuconostoc spp., But are not limited thereto. Preferably, the lactic acid bacteria are Bacillus subtilis or Bacillus polyfermenticus.

Further, in the present invention, the lactic acid bacteria may be contained directly in the composition or in the form of an inclusion. That is, the lactic acid bacteria according to the present invention include at least one of lactic acid bacteria and lactic acid bacteria.

The lactic acid bacteria-containing material may be selected from the group consisting of a suspension of lactic acid bacteria, a culture medium (including lactic acid bacteria, a supernatant, and a medium itself), a lactic acid bacteria broth (obtained by removing solid compounds from a culture medium ), Lactic acid bacteria fermented milk (lactic acid bacteria drink, sour milk, yogurt, etc.).

The present invention is characterized by containing Angelicae gigantis extract and lactic acid bacterium as an active ingredient. By using a mixture of Angelica gigantosa extract and lactic acid bacteria, the antidiarrheal effect is excellent due to the synergistic action of Angelicae gigantis extract and lactic acid bacteria, . Also, when used at a specific blending ratio, the effect is further maximized.

Angelica root extract and lactic bacteria mixing ratio according to the present invention, Angelica extract 5 ~ 30mg / kg: lactic acid 1 × 10 ⁵ ~ 10 ⁹ may be cfu, preferably Angelica Extract 10 ~ 25mg / kg: lactic acid 1 × 10 ⁶ ~ 10 ⁸ cfu, and more preferably from 17 to 23 mg / kg of lactic acid bacteria extract: 1 × 10 ⁶ to 10 ⁸ cfu, but the mixing ratio is not limited thereto. It has been found that the present invention exhibits an optimal anti-inflammatory effect in such a mixing ratio range.

In addition, the present invention can be a mixture of the Angelica gigantosa extract and the lactic acid bacteria culture solution at a weight ratio of 1: 1, 2: 1 or 1: 2, and preferably 1: 1 of the same weight ratio.

In the present invention, the inflammatory diseases are inflammatory bowel disease, Crohn's disease, colitis, inflammatory skin disease, atopic dermatitis, pancreatitis, rheumatoid arthritis, peritonitis, osteomyelitis, encrustitis, dermatitis, allergy, conjunctivitis, periodontitis, rhinitis, otitis, Sjogren's Syndrome, Sjogren's Syndrome, Sjogren's Syndrome, Sjogren's Syndrome, Sjogren's Syndrome, Sjogren's Syndrome, Sjogren's Syndrome, Sjogren's Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome Syndrome syndrome, multiple sclerosis, and the like.

The combined preparation of Angelicae gigantis extract and lactic acid bacterium according to the present invention has no side effects in the human body and is superior in the treatment effect of colitis, such as reduction of body weight, decrease of colonic length and increase of disease activity in an animal model of colitis, However, since the itchiness is improved and the therapeutic effect is on the atopic dermatitis, it can be usefully used in the field of pharmaceuticals and health functional foods for the treatment of inflammatory diseases.

In one aspect, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

The pharmaceutical composition according to the present invention is prepared for the purpose of prevention or treatment of diseases, and can be formulated into various forms according to ordinary methods. For example, it can be formulated into oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions and syrups, and can be formulated in the form of external preparations, suppositories, and sterilized injection solutions.

The pharmaceutical composition of the present invention may be prepared by incorporating one or more pharmaceutically acceptable carriers in addition to the above-mentioned active ingredients for administration. The pharmacologically acceptable carrier to be contained in the pharmaceutical composition of the present invention is one commonly used in the present invention and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, But are not limited to, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. It is not. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above components.

For example, when the pharmaceutical composition of the present invention is formulated into an ointment, cream, or the like, it is possible to use an animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, Talc, zinc oxide or the like as a carrier.

The dosage of the pharmaceutical composition of the present invention may be varied depending on the formulation method, administration method, administration time and / or route of administration of the pharmaceutical composition, and the kind and degree of the reaction to be achieved by the administration of the pharmaceutical composition Including, but not limited to, age, weight, general health, severity or severity of the disease, sex, diet, excretion, drugs used simultaneously or simultaneously with the subject, May be varied according to similar factors well known in the medical arts and those of ordinary skill in the art can readily determine and prescribe dosages that are effective for the desired treatment.

The dosage of the pharmaceutical composition of the present invention may be, for example, 1 mg / kg to 1,000 mg / kg per day, preferably 10 mg / kg to 500 mg / kg per day, And are not intended to limit the scope of the invention.

The route of administration and the mode of administration of the pharmaceutical composition of the present invention may be independent of each other, and the method is not particularly limited, and any route of administration and administration method may be used as long as the pharmaceutical composition can reach the desired site . The pharmaceutical composition may be administered orally or parenterally. The parenteral administration method includes, for example, intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration or subcutaneous administration, and a method of applying, spraying or inhalation the above-mentioned pharmaceutical composition to a disease site But is not limited thereto.

In another aspect, the present invention provides a food composition for preventing or ameliorating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

In the present invention, the health functional food refers to a food group imparted with added value to function or express the function of the food by physical, biochemical or biotechnological techniques, or to control the bio-defense rhythm of the food composition, And restoration, etc. of the body, and the active ingredient according to the present invention may be included in a health functional food for the prevention or improvement of an inflammatory disease.

In the present invention, the food composition may be prepared by a method commonly used in the art, and may be prepared by adding raw materials and ingredients that are conventionally added in the art. The formulations of the food composition may also be prepared without limitation as long as they are formulations acceptable as food compositions.

Foods according to the present invention include, for example, various foods, beverages, gums, tea, vitamin complexes, and functional foods. Foods also include special nutritional foods (eg crude oil, spirits, baby food, etc.), processed meat products, fish meat products, tofu, jelly, noodles (eg, ramie noodles, (For example, soy sauce), candy, chocolate, gum, ice cream, milk products (eg fermented milk, cheese), other processed foods, kimchi, pickled foods (Eg, fruit juices, etc.), beverages (eg fruit drinks, vegetable beverages, beverages, fermented beverages, etc.), natural seasonings (eg, ramen soup, etc.), food additives. The food, beverage or food additive may be prepared by a conventional production method.

When the composition of the present invention is used as a health functional food additive, the composition may be added as it is or may be used together with other health functional food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use. In general, the composition of the present invention may be added in an amount of preferably 50 parts by weight or less, more preferably 25 parts by weight or less, with respect to the raw material in the production of food or beverage. However, in the case of long-term intake intended for health control and hygiene, the amount may be less than the above range, and since there is no problem in terms of stability, the active ingredient may be used in an amount in the above range.

The food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient as well as ordinary food compositions, in addition to the active ingredient Angelicae gigantis extract and lactic acid bacteria. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).

In addition, the food composition may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and aggravating agents (such as cheese and chocolate), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juices and fruit juice drinks and vegetable drinks.

In another aspect, the present invention provides a quasi-drug composition for preventing or ameliorating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

In the present invention, the term "quasi-drug" means a fiber, a rubber product or the like used for the purpose of treating, alleviating, treating or preventing a disease of a human or an animal, a weak action on the human body, Or products similar to those which are not machinery, preparations used for sterilization, insecticides and similar uses for the prevention of infections, for the purpose of diagnosis, treatment, alleviation, treatment or prevention of diseases of human beings or animals Machinery, or apparatus, and that is not an apparatus, machine, or apparatus of an article used for the purpose of giving pharmacological effects to the structure or function of a person or animal, It also includes supplies.

The external preparation for skin is not particularly limited, but may be preferably used in the form of an ointment, a lotion, a spray, a patch, a cream, a powder, a suspension, an gel or a gel.

When the composition of the present invention is added to a quasi-drug composition for the purpose of preventing or ameliorating an inflammatory disease, the composition may be added as it is or may be used together with other quasi-drugs, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use.

In another aspect, the present invention provides a cosmetic composition for preventing or ameliorating inflammatory diseases, including Angelica gigantosa extract and lactic acid bacteria.

In the present invention, the term "cosmedical, cosmeceutical" refers to a product having a professional function which is emphasized on physiologically active effects and effects, unlike general cosmetics, The functional cosmetic refers to a product that helps improve the skin inflammatory disease.

In the present invention, the cosmetic composition may be prepared in any form conventionally prepared, and examples thereof include solutions, emulsions, suspensions, pastes, creams, lotions, gels, powders, sprays, surfactant- , A soap, a liquid cleansing agent, a bath agent, a foundation, a makeup base, an essence, a lotion, a foam, a pack, a soft water, a sunscreen cream or a sun oil.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, or tracant may be used.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The cosmetic composition of the present invention may further comprise customary adjuvants and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, perfumes and the like which are conventionally used. For example, the cosmetic composition may further contain auxiliary ingredients such as glycerin, butyleneglycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate, and allantoin.

The active ingredient according to the present invention may be contained in an amount of 0.01 to 50% by weight, preferably 1 to 30% by weight, based on the total weight of the cosmetic composition of the present invention, but is not limited thereto.

Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples. The following Examples and Experimental Examples are provided only for illustrating the present invention, but the present invention is not limited by the following Examples and Experimental Examples.

Example  1. Preparation of Angelica giganta extract

The truncated angelic roots were dried at 80 ° C to a moisture content of 5% or less. After that, 95% ethanol of 2 to 5 times of the angelic angelic roots was added, and the mixture was refluxed at 80 ° C for 4 hours and then concentrated at 60 ° C using a rotary evaporator. The precipitate produced in the production of the concentrate was removed and a Angelica giganta extract was obtained. Hereinafter, Angelicae Radix extract was named AGNE.

Example  2. Production of lactic acid bacteria

Bacillus polyfermenticus KJS-2 was used as the lactic acid bacteria. The strain has a bacterium shape, and the DNA sequence of 16s rRNA was identified as a spore forming strain. As a result, it was confirmed that the strain was Bacillus polyfermenticus. It also has lactic acid bacteria that consume glucose and produce lactic acid. The above-mentioned Bacillus polyfermenticus KJS-2 was deposited on August 16, 2006 at the Korea Culture Center of Microorganisms (KCCM), and received the deposit number KCCM10769P. After culturing the strain with a fermenter, the cells were collected and spray-dried to prepare a cell powder. Hereinafter, lactic acid bacteria were named as BP2.

Example  3. Preparation of Mixed Material of Angelicae Radix Extract and Lactic Acid Bacteria

In order to prepare a mixture of Angelica gigas Nakai and Lactic acid bacteria, a suspension of Angelica gigas extracts was prepared. The Angelica keiskei koidz. Extract obtained in Example 1 was dissolved in alcohol ethanol to a concentration of 250 mg / ml, and 2 ml of the solution was transferred to a 15 ml Conical tube. Tween 80 and 1 g were added thereto to prepare a primary suspension. Thereafter, 90 ml of purified water was added to a 500 ml bottle, and the mixture was stirred at 600 rpm at 50 ° C. The first suspension was mixed and adjusted to 100 ml with purified water to prepare a secondary suspension (Angelica giganta extract 5 mg / ml) . The solutions were mixed with PBS or physiological saline to give a final dosage level according to Table 1 below.

Next, in the case of lactic acid bacteria, 0.1 mg of the dried powder of BP2 obtained in Example 2 was mixed with Angelica gigantosa extract to give 10 ⁷ CFU per animal.

The specific administration concentrations are shown in Table 1 below, and 0.15 ml of each mixed solution was orally administered to experimental animals.

Group Sample Capacity Lactic acid bacteria Physiological saline solution 1,050 ul + lactic acid bacteria powder 0.1 mg Lactic acid bacteria +
Angelica gigantosa extract 10mg / kg Physiological saline 1,050 ul + lactic acid bacteria powder 0.1 mg + Angelica gigantum extract 5 mg / ml 280 ul Lactic acid bacteria +
Angelica gigantosa extract 20mg / kg Physiological saline 1.050 ul + lactic acid bacteria powder 0.1 mg + Angelica gigantum extract 5 mg / ml 560 ul

Example  4. Preparation of Mixed Material of Angelicae Radix Extract and Lactic Acid Bacteria Culture

BP2 culture medium was prepared by adding 0.1 g (1 x 10 ¹⁰ cfu / g) of BP2 powder to 10 mL of sterilized distilled water and then suspending. 0.1 mL of the suspension was added to the Tryptic soy broth medium and cultured at 37 ° C in a shaking incubator at 150 rpm for 3 days.

The concentrations of Angelica gigas Nakai extract and BP2 medium (10 ⁷ CFU) were 0.01, 0.1 and 1 mg / mL, respectively, and the mixture ratio of Angelica gigas Nakai extract and BP2 medium was 1: 1. Respectively.

Experimental Example  1. Validation of treatment effect in colitis animal model

1-1. Manufacture of animal models

Four week old male ICR mice (Daehan biolink, Korea) were purchased and used in the experiment after purification. The incubation room was maintained at a temperature of 22 ± 2 ° C and a humidity of 50 ± 5%, and the photoperiod and cancer cycle were adjusted to 12 hours. Animal testing was approved by the Daegu Han Institute of Animal Care and Use Committee (IACUC) (approval number: DHU2016-045).

In order to induce colitis, DSS (MP biomedicals, USA) with a molecular weight of 36,000 ~ 50,000 was diluted to 5% in tap water prior to experiment and freely taken for 9 days in colitis inducing group and drug administration group. Respectively. In the colitis - induced group and the drug - treated group, the positive control group SSZ (sulfasalazine 300 mg / kg administered) was orally administered once a day from the day of DSS supply, and the body weight and disease activity level were measured daily. In the case of the drug-administered group, as shown in Example 3, the Angelica giganta extract was diluted in 0.9% saline and orally administered (0.1 mg / kg or 20 mg / kg) to the experimental animals One BP2 was administered together at a concentration of 10 < ⁷ > CFU.

1-2. Weight change analysis

The body weight change was measured during the experiment, and the results are shown in FIG.

As shown in Fig. 1, the weight change of the control group consumed with tap water was continuously increased, but the weight of the group ingesting DSS (colitis induced group / negative control group) increased until 5th day. Respectively. On the other hand, the combined administration of AGNE and BP2 according to the present invention showed an increase in body weight and an insignificant decrease after 5 days.

1-3. Colon length analysis

In order to confirm the therapeutic effect of colitis, the length of the colon was measured after the experiment. The results are shown in Fig.

As shown in FIG. 2, the colon length of the control group consumed tap water was 10.2 ± 0.2 cm, but the colon length of the group ingesting DSS (colitis induced group / negative control group) was significantly decreased to 5.5 ± 0.3 cm. In contrast, the combined administration of AGNE and BP2 according to the present invention showed 6.8 ± 0.2 and 8.5 ± 0.3 cm, respectively, and the increase in the colonic length was dependent on the dose of AGNE. Particularly, the concentration of AGNE was increased to 20 mg / kg (7.5 ± 0.5 cm) in the group treated with SSZ as a positive control group.

1-4. Degree of disease activation  (Disease activity index; DAI ) Measure

The score was measured by measuring weight loss, stool, and anal status due to colitis according to Table 2 below. The body weight loss was calculated from the weight loss rate on the first and last days of the experiment, and the state of the sides and anus were calculated by summing up the state of the last day. The medical variable used here is a comprehensive functional measure similar to the medical symptoms of ulcerative colitis in the human body. The results are shown in Fig.

[Table 2]

As shown in FIG. 3, no DAI was observed in the Control group ingesting tap water, but the DSS intake group (colitis induced group / negative control group) was significantly increased to 5.8 ± 0.3. In contrast, the combined administration of AGNE and BP2 according to the present invention showed significant DAI reduction of 3.8 ± 0.3 and 3.3 ± 0.3, respectively.

Experimental Example  2. Verification of treatment effect in animal model of atopic dermatitis

2-1. Manufacture of animal models

Four-week-old male BALB / C mice (Daehan biolink, Korea) were purchased and used for the experiment after purification. The incubation room was maintained at a temperature of 22 ± 2 ° C and a humidity of 50 ± 5%, and the photoperiod and cancer cycle were adjusted to 12 hours. Animal testing was approved by Daegu Han's Institutional Animal Care and Use Committee (IACUC) (approval number: DHU2016-044).

To induce itching, the ICR mouse was orally administered drug and physiological saline, and each of the mice was placed into a transparent acrylic cage for 30 minutes, followed by 30 minutes of adaptation. Subsequently, copound 48/80 (15 ug / site) was subcutaneously injected between both shoulders such as a mouse, and the number of times of scraping the area injected with the hind paw for 1 hour was measured.

In order to induce atopic dermatitis, the back part of the mouse was depilated, and then 2,4-dinitrochlorobenzene (DNCB, Sigma, USA) was dissolved in acetone and olive oil (3: 1) And 200 [mu] L of the solution was applied to the back region for 3 days. Two days after inducing the primary immune response, DNCB diluted to 0.5% was applied every other day. After 2 weeks, the drug was administered orally, followed by application of DNCB to inhibit natural healing.

2-2. Itching assessment

In order to confirm the therapeutic effect on dermatitis, itching was evaluated and the results are shown in Fig.

As shown in Fig. 4, the rats in the control group were scratched 9.8 ± 3.0 times, but those in the compound 48/80 group were scratched 119.4 ± 7.3 times. On the other hand, in the combined treatment of AGNE and BP2 according to the present invention, it was confirmed that the number of times of scratching remarkably decreased by 70.2 ± 5.2 times and 56.0 ± 3.3 times.

2-3. Measure skin damage

Clinical skin severity score of the experimental animals was measured 5 weeks after the oral administration of the drug by the clinical visual evaluation method generally used for atopic dermatitis and the values were summed up. (1), mild (1), severe (2), and hyperhidrosis (3). In the case of DNCB solution, the experimental animal has erythema / pigmentation, exudate / dirt, scratch, . The results are shown in Fig.

As shown in FIG. 5, normal skin control group was not able to observe skin damage, but DNCB group was 10.0 ± 1.1, which caused significant skin damage. On the other hand, the combination of AGNE and BP2 according to the present invention showed 5.1 ± 0.7 and 4.4 ± 0.8, respectively. This was superior to the Terfenadine group used as a positive control.

Experimental Example  3. Determination of cytokine production of mixed material of Angelicae Radix extract and lactic acid bacteria culture

RAW 264.7 cells were pretreated with various concentrations of the sample and stimulated with LPS for 24 hours. TNF-α and IL-1β levels were measured in the culture supernatant. The cytokine concentration was measured by enzyme-linked immunosorbent assay (ELISA) and the method was as follows. The monoclonal antibody against each cytokine was diluted with carbonate coating buffer (Na ₂ CO ₃ / NaHCO ₃ ) to a 96-well plate at a pH of 9.5 and coated on a 96-well plate at a rate of 100 μl each. Respectively. The plate was washed with PBS containing 0.05% tween, and then blocked with PBS containing 1% BSA, 5% sucrose and 0.05% NaN ₃ for 1 hour. After washing several times, the samples were added and incubated at 37 ° C for 2 hours. Each well was washed again, and the biotin-conjugated secondary antibody was added and left for another 2 hours. After washing the wells, AP was added and allowed to stand at 37 ° C for 30 minutes, then the wells were washed again, and the substrate ABTS solution was added. The color development reaction was measured at 405 nm using an ELISA reader. The concentration of Angelica gigas Nakai extract was 0.01, 0.1 and 1 mg / mL. The BP2 culture medium was used as the undiluted solution. The mixture ratio of Angelica gigas extract and BP2 culture was 1: 1. The results are shown in Fig. 6 and Fig.

As shown in FIG. 6, it was confirmed that the amount of IL-1β produced by LPS stimulation was 6.01 ± 0.14 ng / mL, which was significantly higher than that of the control (2.85 ± 0.1) IL-1β production was significantly inhibited compared to the LPS-treated group.

In addition, as shown in FIG. 7, it was confirmed that the amount of TNF-α produced by LPS stimulation was 7.18 ± 0.13 ng / mL, which was significantly higher than that of the control (0.77 ± 0.07) TNF-α production was significantly inhibited by the LPS-treated group.

Inflammatory cytokines play a role in controlling various immune and inflammatory responses in the body. The macrophage stimulated by LPS of the bacterium produces TNF-α, and the secreted TNF-α and LPS induce the production of IL-1β, thereby continuing the inflammatory response. TNF-a induced by LPS promotes the onset of the inflammatory response and continuous production leads to chronic inflammation and eventually to various physiological processes such as septic shock, inflammation and cytotoxicity.

These results indicate that a mixture of Angelica gigantis extract and BP2 culture medium acts at the early stage of the inflammatory cytokine mechanism and is most effective at inhibiting inflammatory cytokines.

The results of the above experiments show that the combined treatment of Angelicae gigantis extract and lactic acid bacterium is superior to the treatment of colitis in animal models of colitis, such as reduction of body weight, decrease of colonic length and increase of disease activity, It was confirmed that the itching was improved and the therapeutic effect on atopic dermatitis was excellent.

Hereinafter, a pharmaceutical composition, a food composition and a cosmetic composition containing the composition of the present invention will be described. However, the present invention is not intended to be limited thereto but is specifically described below.

Formulation example  1. Preparation of pharmaceutical compositions

1- 1. Sanje  Produce

Angelica gigantum extract and lactic acid bacteria 2 g

Lactose 1 g

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

1- 2. Preparation of tablets

Angelica gigantum extract and lactic acid bacteria 100 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

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

1-3. Preparation of capsules

Angelica gigantum extract and lactic acid bacteria 100 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

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

Formulation example  2. Preparation of food composition

2-1. Manufacture of Health Functional Foods

Angelica gigantum extract and lactic acid bacteria 100 mg

Vitamin mixture quantity

Vitamin A acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

0.15 mg of vitamin B2

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

Biotin 10 μg

Nicotinic acid amide 1.7 mg

Folic acid 50 μg

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg of ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Secondary calcium phosphate 55 mg

Potassium citrate 90 mg

Calcium carbonate 100 mg

Magnesium chloride 24.8 mg

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

Formulation example  3. Cosmetics  Preparation of composition

3-1. Manufacture of softening longevity (skin lotion)

Angelica sinensis extract and lactic acid bacteria 0.5%

Beta-1,3-glucan 1.0%

Butylene glycol 2.0%

Propylene glycol 2.0%

Carboxyvinyl polymer 0.1%

Phage-12 nonyl phenyl ether 0.2%

Polysorbate 80 0.4%

Ethanol 10.0%

Triethanolamine 0.1%

Preservative, pigment, perfume

Purified water to 100%

3-2. Nutrition lotion ( Milk lotion )

Angelica sinensis extract and lactic acid bacteria 0.5%

Beta-1,3-glucan 1.0%

Waste 4.0%

Polysorbate 60 1.5%

Sorbitan sesquioleate 1.5%

Liquid paraffin 0.5%

Caprylic / capric triglyceride 5.0%

Glycerin 3.0%

Butylene glycol 3.0%

Propylene glycol 3.0%

Carboxyvinyl polymer 0.1%

Triethanolamine 0.2%

Preservative, pigment, perfume

Purified water to 100%

3-3. Manufacture of nutrition cream

Angelica sinensis extract and lactic acid bacteria 1.0%

Beta-1,3-glucan 5.0%

Wax 10.0%

Polysorbate 60 1.5%

Pigment 60 Hardened castor oil 2.0%

Sorbitan sesquioleate 0.5%

Liquid paraffin 10.0%

Squalane 5.0%

Caprylic / capric triglyceride 5.0%

Glycerin 5.0%

Butylene glycol 3.0%

Propylene glycol 3.0%

Triethanolamine 0.2%

Preservative, pigment, perfume

Purified water to 100%

Korea Microorganism Conservation Center (Domestic) KCCM10769P 20060816

Claims (8)

A pharmaceutical composition for preventing or treating an inflammatory disease, comprising Angelica gigantosa extract and lactic acid bacteria. [Claim 2] The pharmaceutical composition according to claim 1, wherein the Angelica keiskei koidz. Extract is extracted with at least one solvent selected from water, alcohols having 1 to 4 carbon atoms, and a mixed solvent thereof. The method of claim 1, wherein the lactic acid bacteria are selected from the group consisting of Bacillus, Lactobacillus, Streptococcus, Bifidobacterium, Lactococcus, Pediococcus, ) And Leuconostoc genus. ≪ Desc / Clms Page number 15 > The pharmaceutical composition according to claim 3, wherein the lactic acid bacteria are Bacillus subtilis or Bacillus polyfermenticus. The method according to claim 1, wherein the inflammatory disease is inflammatory bowel disease, Crohn's disease, colitis, inflammatory skin disease, atopic dermatitis, pancreatitis, rheumatoid arthritis, peritonitis, osteomyelitis, colitis, dermatitis, allergy, conjunctivitis, periodontitis, rhinitis, Sphincter, tonsillitis, pneumonia, gastric ulcer, gastritis, hemorrhoids, gout, ankylosing spondylitis, lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, shoulder circumfection, tendinitis, hay fever, tendinitis, myositis, hepatitis, cystitis, sjogren ' s syndrome, and multiple sclerosis. < Desc / Clms Page number 24 > A food composition for preventing or ameliorating an inflammatory disease comprising Angelica gigas Nakai extract and Lactic acid bacteria. A quasi-drug composition for preventing or ameliorating an inflammatory disease including Angelica gigas Nakai extract and lactic acid bacteria. A cosmetic composition for preventing or improving an inflammatory disease, including Angelica gigantosa extract and lactic acid bacteria.

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