KR102578112B1 - A composition for preventing or treating inflammatory disease comprising extract of C. hassaku Hort. ex Y. Tan and Rehmanniae Radix preparata as effective component - Google Patents

Abstract

The present invention relates to a pharmaceutical composition, food composition, feed composition, quasi-drug composition, and cosmetic composition for the prevention or treatment of inflammatory diseases, containing a mixed extract of citrus and Rehmannia glutinosa as an active ingredient.

Description

A composition for preventing or treating inflammatory disease comprising extract of C. hassaku Hort. ex Y. Tan and Rehmanniae Radix preparata as effective component}

The present invention relates to pharmaceutical compositions, food compositions, feed compositions, quasi-drug compositions, and cosmetic compositions for the prevention or treatment of inflammatory diseases, containing mixed extracts of Red Palsy and Rehmannia glutinosa as active ingredients.

Inflammatory bowel disease (IBD) is a disease that causes chronic inflammation in the gastrointestinal tract. It occurs relatively young and is accompanied by symptoms such as abdominal pain, fever, diarrhea, and bleeding. Broadly, it is classified into two types: ulcerative colitis (UC) and Crohn's disease (CD). Ulcerative colitis mainly invades the mucous membrane and frequently causes ulcers or ulcers in the large intestine of unknown cause. It is a type of diffuse nonspecific inflammation and is accompanied by various systemic symptoms, including bloody diarrhea. Crohn's disease is a discontinuous condition of the entire digestive tract from the mouth to the anus, causing ulcers in all layers of the intestinal tract from the mucosa to the anus. It is a granulomatous inflammatory lesion of unknown cause that progresses into fibrosis, stenosis, and lesions, and is accompanied by systemic symptoms such as abdominal pain, chronic diarrhea, fever, and nutritional disorders.

Recently, in order to effectively relieve inflammation, research is being conducted on substances that can inhibit the production of NO. However, anti-inflammatory substances developed through these studies have some side effects.

In order to treat these inflammatory bowel diseases, research is being actively conducted to develop compositions that can treat inflammatory bowel diseases from natural products with fewer side effects. For example, Korean Patent No. 1466308 discloses a composition for the treatment of colitis containing an extract of Hypophyllosis as an active ingredient, and Korean Patent No. 1526467 discloses a composition for the treatment of colitis containing Cornus officinalis extract as an active ingredient. . However, these extracts derived from natural products have limitations in that their effectiveness in preventing or treating colitis is not satisfactory.

Accordingly, the present inventors have made extensive research efforts to develop a natural product-derived material that has anti-inflammatory effects and has fewer side effects. As a result, it has been confirmed that the Hongpalsac extract and the Rehmannia glutinosa extract are effective in preventing or treating inflammatory bowel disease, thereby providing the present invention. Completed.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory diseases containing a mixed extract of Red Palsy and Rehmannia glutinosa as an active ingredient.

Another object of the present invention is to provide a method for preventing or treating inflammatory diseases comprising administering the composition to an entity other than a human.

One object of the present invention is to provide a food composition for preventing or improving inflammatory diseases, which contains a mixed extract of Red Palsy and Rehmannia glutinosa as an active ingredient.

One object of the present invention is to provide a feed composition for preventing or improving inflammatory diseases containing a mixed extract of Red Palsac and Rehmannia glutinosa as an active ingredient.

Another object of the present invention is to provide a quasi-drug composition for preventing or improving inflammatory diseases containing a mixed extract of Red Palsy and Rehmannia glutinosa as an active ingredient.

Another object of the present invention is to provide a cosmetic composition for alleviating or improving inflammation containing a mixed extract of Red Pale Root and Rehmannia glutinosa as an active ingredient.

Hereinafter, the contents of the present invention will be described in detail as follows. Meanwhile, the description and embodiment of one aspect disclosed in the present invention may also be applied to the description and embodiment of other aspects with respect to common matters. Additionally, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. In addition, the scope of the present invention cannot be considered limited by the specific description described below.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases, comprising a mixed extract of Red Palsy and Rehmannia glutinosa as an active ingredient.

The term "C. hassaku Hort. ex Y. Tan" in the present invention is one of the citrus varieties and is an azo mutation of Palsak, a Japanese citrus variety. It is currently unknown whether Hongpalsac extract has an anti-inflammatory effect. Tangerines are a well-known resource to consumers and contain a large amount of useful ingredients such as flavonoids and carotenoids, so they have the advantage of being able to be developed as a material to improve inflammatory colitis.

The term "Rehmanniae Radix preparata" in the present invention is a medicine made from the roots of Rehmanniae Radix, a perennial medicinal plant, and is a medicine made by repeating the process of steaming and drying the Rehmanniae roots nine times. The root of Rehmannia glutinosa originally has a cold nature, but Suk Rehmannia glutinosa loses its cold nature and exhibits a warm nature and sweet taste.

In the present invention, the mixed extract of Hongpalsak and Rehmannia glutinosa may be interchangeably named 'HSE'. The mixed extract may be an extract of a mixture of Red Palsy and Rehmannia glutinosa, and may also be a mixture of Red Palsy and Rehmannia glutinosa extracts.

In the present invention, the Hongpalsak and Rehmannia glutinosa can be purchased and used commercially, or those collected or cultivated in nature can be used, but are not limited thereto.

The term "extract" in the present invention may refer to a result such as a liquid component obtained by immersing in a solvent and then extracting the extract at room temperature or at a heated state for a certain period of time, or a solid component obtained by removing the solvent from the liquid component.

The above-mentioned Hongpalsac extract and Rehmannia glutinosa extract, which have inflammatory response inhibitory activity, can be extracted from various organs of natural, hybrid, and mutated plants, such as roots, stems, leaves, flowers, fruit bodies, and fruit peels, as well as plants. It can be extracted from tissue culture, but is not limited thereto.

In the present invention, the Hong Palsak extract and the Rehmannia glutinosa extract include, but are not limited to, those extracted with a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.

In the present invention, the extract may include any of the following: an extract obtained by extraction treatment, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof.

The Hongpalsak and Rehmannia glutinosa extracts can be prepared using general extraction, separation and purification methods known in the art. The extraction method is not limited to this, but specifically, boiling water extraction, hot water extraction, cold needle extraction, reflux cooling extraction, or ultrasonic extraction can be used.

In the present invention, the mixture of Hongpalsac and Rehmannia glutinosa has a weight ratio (w/w) of 4.5:0.5, a weight ratio of 4.0:1.0, a weight ratio of 3.0:0.5, a weight ratio of 3.0:1.0, and a weight ratio of 2.5:0.5. , may include a weight ratio of 2.0:1.0, a weight ratio of 2.0:0.5, a weight ratio of 1.0:1.0, a weight ratio of 1.0:0.5, specifically a weight ratio of 1.0 to 3.0:1.0, and more specifically a weight ratio of 3.0:1.0. It can be done, but is not limited to this.

Additionally, the composition may further include citrus biogel, but is not limited thereto.

The term "citrus biogel" of the present invention refers to a material made by culturing microorganisms in citrus juice, and can be used interchangeably with citrus cellulose gel. Citrus biogel has a uniform structure and many gaps, so it has good conditions for adsorbing new substances, so it has the advantage of being industrialized as functional cosmetics and pharmaceutical materials by combining various materials.

The term 'adsorption' of the present invention refers to the phenomenon in which the target component gathers on the surface of biogel and/or cellulose gel. Due to the influence of the pores of the biogel and/or cellulose gel, the target component may appear to be gathered on the surface of the area where the pores are formed, so the target component may be included inside the cellulose gel, but is limited to this. It doesn't work. Specifically, biogel refers to a biogel manufactured using citrus juice or by-products remaining after citrus juice. For example, the remaining residue remaining after citrus juice or citrus juice is used as a medium and glued to the medium. It may be a biogel produced by inoculating with Gluconacetobacter sp. gel_SEA623-2 (KACC 91526P), but is not limited thereto. The citrus biogel is mainly composed of insoluble dietary fiber. Insoluble dietary fiber is excellent for absorbing moisture present in the digestive tract, increasing the volume of stool, stimulating the intestines, and promoting intestinal movement, and excreting harmful substances in the intestine along with the stool. In the present invention, the diversity of beneficial microorganisms in the intestines increases by including citrus biogel in the extracts of Hongpalsac and Rehmannia glutinosa.

Additionally, for the purpose of the present invention, Hongpalsak, one of the citrus varieties, can be used as the tangerine, but is not limited thereto.

In the present invention, the citrus biogel added to the mixed extract of Hongpalsac and Rehmannia glutinosa may be interchangeably named 'HSB'.

In the present invention, the citrus biogel may contain 1 to 30% by weight, 5 to 15% by weight, and 7 to 12% by weight, and specifically 10% by weight, based on the total weight of the composition. Not limited.

As used herein, the term "inflammatory disease" is a general term for diseases whose main lesion is inflammation, for example, inflammatory skin disease, allergic disease, Crohn's disease (Crohn's disease), inflammatory bowel disease such as ulcerative colitis, and peritonitis. , osteomyelitis, cellulitis, meningitis, encephalitis, pancreatitis, trauma-induced shock, bronchial asthma, cystic fibrosis, stroke, acute bronchitis, chronic bronchitis, acute bronchiolitis, chronic bronchiolitis, osteoarthritis, gout, spondyloarthropathy, ankylosing spondylitis, Reiter syndrome, Psoriatic arthropathy, enteropathy spondylitis, juvenile arthropathy, juvenile ankylosing spondylitis, reactive arthropathy, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, syphilitic arthritis, Lyme. Disease, arthritis associated with 'vasculitis syndrome', polyarteritis nodosa, hypersensitivity vasculitis, Lou Gehrig's granulomatosis, polymyalgia rheumatica, articular cell arteritis, calcium crystal deposition arthropathy, pseudogout, non-articular rheumatism, bursitis, tenosynovitis, epicondylitis (tennis elbow), neuropathic joint disease (charcoal and joint), hemorrhagic arthrosis (hemarthrosic), Henoch-Schönlein purpura, hypertrophic osteoarthropathy, multicentric reticulohistiocytoma, surcoilosis, hemochromatosis, sickle cell and other hemochromatosis, hyperlipoproteinemia, hypogammaglobulinemia, familial Mediterranean fever, Behart's disease, systemic lupus erythematosus, relapsing fever, psoriasis, multiple sclerosis, sepsis, septic shock, multi-organ dysfunction syndrome, acute respiratory syndrome. It may be any one selected from the group consisting of distress syndrome, chronic obstructive pulmonary disease, acute lung injury, and broncho-pulmonary dysplasia, and specifically, inflammatory skin disease. , ulcerative colitis, chronic bronchitis, osteoarthritis, and sepsis. It is not limited to this.

The term “prevention” in the present invention refers to all actions that suppress or delay inflammatory diseases by administering the composition according to the present invention.

The term “treatment” in the present invention refers to any action in which the inflammatory disease is improved or beneficially changed by administration of the composition according to the present invention.

In a specific embodiment of the present invention, as a result of treating a colitis animal model with HSE (Rhododendron chinensis + Rehmannia glutinosa + Rehmannia glutinosa + Rehmannia glutinosa + Tangerine biogel), the colon length in both treatment groups was similar to that of the normal group. It was confirmed that the patient recovered well, and mucous membrane damage was also recovered. In particular, it was confirmed that the HSB group was more effective in improving enteritis (Figures 5 and 6).

In another specific example of the present invention, it was confirmed that short-chain fatty acids, which help promote regeneration of the damaged colon, were increased in both the HSE and HSB treated groups of the present invention (FIGS. 7 and 8). In addition, it was confirmed that microorganisms related to short-chain fatty acid production tend to gradually recover (FIG. 9). In addition to the above results, it was confirmed that the diversity and homogeneity of microbiota increased when treated with the extracts of the present invention (HSE and HSB) (FIG. 10).

In another specific embodiment of the present invention, as a result of measuring the secretion amount of inflammatory cytokines IL-6 and TNF-α in both the HSE and HSB treated groups of the present invention, the secretion amount of inflammatory cytokines was significantly A decrease was confirmed (Figure 11). This suggests that treatment with HSE and HSB may have a therapeutic effect on inflammatory diseases by reducing the secretion of inflammatory cytokines that are directly involved in the inflammatory response.

The term “pharmaceutical composition” used in the present invention refers to a product prepared for the purpose of preventing or treating diseases, and can be formulated and used in various forms according to conventional methods. For example, it can be formulated into powders, granules, tablets, capsules, suspensions, emulsions, syrups, etc., and can be formulated and used in the form of external preparations, suppositories, and sterile injection solutions. Specifically, it can be formulated and used in a form suitable for eye administration, for example, eye drops, cream, ointment, gel, or lotion.

The composition of the present invention may be prepared in the form of a pharmaceutical composition for preventing or treating inflammatory diseases, which further comprises an appropriate carrier, excipient, or diluent commonly used in the manufacture of pharmaceutical compositions. The carrier may be a non-natural carrier. (non-naturally occurring carrier) may be included.

In the present invention, carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Examples include calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain the leaf extract and its fractions with at least one excipient, such as starch or calcium carbonate. It is prepared by mixing sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate and talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, and syrups, and may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives in addition to the commonly used simple diluents such as water and liquid paraffin. there is. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurin, glycerogeratin, etc. can be used.

The pharmaceutical composition of the present invention can be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, etc., depending on the desired method, and the active ingredient of the pharmaceutical composition is determined by the age of the recipient. , gender, weight, pathological condition and its severity, route of administration, or the judgment of the prescriber. Determination of the application amount based on these factors is within the level of one skilled in the art, and the daily dosage is, for example, 0.1 mg/g/day to 100 mg/g/day, more specifically 5 mg/g/day to 50 mg/day. g/day, and the frequency of administration of the composition of the present invention is not particularly limited, but may be administered once a day or divided into multiple doses. The above dosage does not limit the scope of the present invention in any way.

Another aspect of the present invention provides a method for preventing or treating inflammatory diseases, comprising administering the composition to an entity other than a human.

The Hongpalsak and Rehmannia glutinosa extracts, inflammatory diseases, and prevention are as described above.

The term “individual” in the present invention may refer to any animal, including humans, that has developed or is likely to develop an inflammatory disease. The animal may be not only a human, but also a mammal such as a cow, horse, sheep, pig, goat, camel, antelope, dog, or cat that requires treatment for similar symptoms, but is not limited thereto.

The prevention or treatment method of the present invention may specifically include administering the composition in a pharmaceutically effective amount to an individual who has developed or is at risk of developing an inflammatory disease.

The term "administration" in the present invention means introducing the pharmaceutical composition of the present invention into a patient by any appropriate method, and the route of administration of the composition of the present invention may be administered through various routes as long as it can reach the target tissue. Depending on the purpose, it can be administered through routes such as eye drop administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, and intrarectal administration. It can be administered, and specifically, it can be administered through the route of oral administration.

One aspect of the present invention for achieving the above object provides a food composition for preventing or improving inflammatory diseases, comprising a mixed extract of Red Palsy and Rehmannia glutinosa as an active ingredient.

The term "improvement" in the present invention refers to any action in which the symptoms of an inflammatory disease are improved or beneficially changed by administering the composition.

The food composition of the present invention provides various types of food additives or functional foods. Specifically, it can be processed into leached tea, liquid tea, beverage, fermented milk, cheese, yogurt, juice, probiotics or health supplements containing the composition, and can be used in the form of various other food additives.

The food composition of the present invention may further include a foodologically acceptable carrier.

There is no particular limitation on the type of food to which the composition containing the Hongpalsac extract and the Rehmannia glutinosa extract of the present invention can be added, for example, various beverages, gum, tea, vitamin complexes, health supplements, etc. Other ingredients that do not interfere with the effect of treating menopausal disorders can be added to the food composition, and their types are not particularly limited. For example, like regular foods, it may contain various herbal extracts, foodologically acceptable food supplements, or natural carbohydrates as additional ingredients.

The food auxiliary additives are added to prepare food compositions of each dosage form and can be appropriately selected and used by those skilled in the art. For example, various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters. , stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated drinks, etc., but the types are not limited to the above examples.

At this time, the content of the extract included in the food is not particularly limited, but may be included in 0.01 to 100% by weight, specifically 1 to 80% by weight, based on the total weight of the food composition.

If the food is a beverage, it may be included at a rate of 1 to 30 g, specifically 3 to 20 g, based on 100 ml. Additionally, the composition may contain additional ingredients that are commonly used in food compositions to improve odor, taste, vision, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. Additionally, it may contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu). Additionally, it may contain amino acids such as lysine, tryptophan, cysteine, and valine. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene ( BHT), etc.), colorants (tar color, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG monosodium glutamate, etc.), sweeteners (dulcine, cyclemate, saccharin, sodium, etc.) , food additives such as flavorings (vanillin, lactones, etc.), leavening agents (alum, D-potassium hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (grease), coating agents, gum base agents, anti-foam agents, solvents, improvers, etc. ) can be added. The additives are selected according to the type of food and used in an appropriate amount.

The food composition of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and components commonly added in the art. In addition, unlike general drugs, it is made from food, so it has the advantage of not having any side effects that may occur when taking the drug for a long time, and it can be highly portable.

Another aspect of the present invention provides a feed composition for preventing or improving inflammatory diseases, comprising a mixed extract of Red Palsy and Rehmannia glutinosa as an active ingredient.

The composition for feed may include feed additives . The feed additive of the present invention corresponds to supplementary feed under the Feed Management Act.

In the present invention, the term "feed" means any natural or artificial diet, meal, etc., or a component of the meal, for or suitable for eating, ingestion, and digestion by animals.

The type of feed is not particularly limited, and feed commonly used in the art can be used. Non-limiting examples of the feed include plant feed such as grains, roots and fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, gourds or grain by-products; Examples include animal feeds such as proteins, inorganic substances, fats and oils, minerals, oils and fats, single-cell proteins, zooplanktons or foods. These may be used alone or in combination of two or more types.

Another aspect of the present invention provides a quasi-drug composition for preventing or improving inflammatory diseases, comprising a mixed extract of Red Palsy and Rehmannia glutinosa as an active ingredient.

The term "quasi-drug" in the present invention refers to products that have a milder effect than pharmaceuticals among products used for the purpose of diagnosing, treating, improving, alleviating, treating, or preventing diseases in humans or animals. For example, quasi-drugs under the Pharmaceutical Affairs Act exclude products used for pharmaceutical purposes and include products used to treat or prevent diseases in humans and animals, and products that have a mild or no direct effect on the human body.

Specifically, the quasi-drugs may include external skin preparations and personal hygiene products. More specifically, it may be a disinfectant cleaner, shower foam, garnish, wet tissue, detergent soap, hand wash, or ointment, but is not limited thereto.

When using the composition according to the present invention as a quasi-drug additive, the composition can be added as is or used together with other quasi-drugs or quasi-drug components, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use.

Another aspect of the present invention provides a cosmetic composition for alleviating or improving inflammation, comprising a mixed extract of Red Palsy and Rehmannia glutinosa as an active ingredient.

In the present invention, the cosmetic composition includes a solution, external ointment, cream, foam, nutritional lotion, softening lotion, pack, softening water, emulsion, makeup base, essence, soap, liquid cleanser, bath salt, sunscreen cream, sun oil, Can be prepared in a formulation selected from the group consisting of suspensions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays. Not limited.

The cosmetic composition of the present invention may further include one or more cosmetically acceptable carriers that are blended with general skin cosmetics, and common ingredients include, for example, oil, water, surfactant, moisturizer, lower alcohol, and thickener. , chelating agents, pigments, preservatives, fragrances, etc. may be appropriately mixed, but are not limited thereto.

Cosmetically acceptable carriers included in the cosmetic composition of the present invention vary depending on the formulation of the cosmetic composition.

When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. are used as carrier ingredients. may be used, but is not limited thereto. These may be used individually or in combination of two or more types.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silcate, polyamide powder, etc. can be used as carrier ingredients, and especially in the case of a spray, chlorofluorohydro It may include, but is not limited to, propellants such as carbon, propane/butane or dimethyl ether. These may be used individually or in combination of two or more types.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butyl glycol oil, etc. can be used, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan. may be used, but is not limited thereto. These may be used individually or in combination of two or more types.

When the formulation of the present invention is a suspension, the carrier components include water, liquid diluents such as ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, and miso. Crystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used, but are not limited thereto. These may be used individually or in combination of two or more types.

When the formulation of the present invention is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, and fatty acid amide. Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linoline derivative, or ethoxylated glycerol fatty acid ester may be used, but are not limited thereto. These may be used individually or in combination of two or more types.

In addition to the above-mentioned essential ingredients, the cosmetic composition of the present invention may contain other ingredients normally blended in cosmetics, if necessary. Specifically, the ingredients that may be added include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohols, pigments, Fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, etc. may be included, but are not limited thereto.

The mixed extract of Hongpalsak and Rehmannia glutinosa provided in the present invention is a material derived from natural products, and has the effect of improving inflammatory colitis, which is increasing in incidence due to various factors such as environmental pollution and stress, while having fewer side effects. Expansion of consumption and improvement of national health can be expected.

Figure 1 is a diagram of strains and products related to citrus biogel production.
Figure 2 is a graph showing the results of cytotoxicity analysis of Red Palsy extract, Rehmannia glutinosa extract, and mixed extract (including ratios) of Red Palsy extract and Rehmannia glutinosa extract.
Figure 3 is a graph comparing the cytokine secretion amount of Hongpalsac extract, Rehmannia glutinosa extract, and mixed extract of Hongpalsac and Rehmannia glutinosa extract (including ratios).
Figure 4 is a graph showing the DAI values of the group treated with HSE and HSB in samples prepared after inducing colitis with DSS.
Figure 5 is a diagram confirming the length of the large intestine of the group treated with HSE and HSB in a sample prepared after inducing colitis with DSS.
Figure 6 is a graph showing the histopathological results of a group treated with HSE and HSB on samples prepared after inducing colitis with DSS.
Figure 7 is a graph analyzing short-chain fatty acids in mouse feces from a group treated with HSE and HSB in samples prepared after inducing colitis with DSS.
Figure 8 is a diagram analyzing the distribution of microorganisms related to short-chain fatty acid production in mouse feces of a group treated with HSE and HSB in samples prepared after inducing colitis with DSS.
Figure 9 is a diagram analyzing the increase in microorganisms related to short-chain fatty acid production in the group treated with HSE and HSB in samples prepared after inducing colitis with DSS.
Figure 10 is a diagram analyzing the diversity and homogeneity of Microbiota in the group treated with HSE and HSB in samples prepared after inducing colitis with DSS.
Figure 11 is a graph confirming whether the secretion of inflammatory cytokines was reduced in the group treated with HSE and HSB in samples prepared after inducing colitis with DSS.

Hereinafter, the present invention will be explained in detail by the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited by the following examples.

Example 1: Preparation and toxicity confirmation of mixed extract

1-1: Production of citrus biogel

Citrus biogel is from Gluconacetobater sp. It was produced using the gel_SEA623-2 (KACC91526P) strain. To produce the citrus biogel, the Gluconacetobacter genus SEA623-2 strain was grown at 1Х10 ⁵ cells/ml in a medium containing Onju tangerine juice (10% (v/v)) with a sugar content of 10 brix and an acidity pH of 3.5. were inoculated and cultured under aerobic conditions at 25°C for 20 days.

As a result, the initial gel production rate of the SEA623-2 strain was so excellent that a gel film began to form from the 2nd day of culture, and the citrus biogel produced after 20 days was recovered and used (Figure 1). In addition, it was confirmed that the citrus biogel used in the present invention is soft, has a three-dimensional network structure, and can be made into nanoparticles with a spacing between fiber bundles of 10-20 nm.

1-2: Preparation of extract

The extracts were extracted by immersing Hongpalsak and Rehmannia glutinosa in 70% ethanol, respectively, and then mixed in various ratios. After preparing the mixed extract, citrus biogel was used by adding 10% by weight of the total composition. The citrus biogel used in the present invention was pulverized to the extent that no foreign body sensation was left when ingested.

1-3: Cytotoxicity evaluation

The inflammatory bowel disease treatment effect of the extracts of each single extract (Rhododendron sinensis extract, Rehmannia glutinosa extract), HSE (Rhododendron sinensis extract + Rehmannia glutinosa), and HSB (Rich persimmon + Rehmannia glutinosa + citrus biogel) prepared in Example 1-2 was examined. Prior to confirmation, a cytotoxicity evaluation was performed to confirm a concentration that was not toxic to cells.

Cytotoxicity was evaluated using the MTT method. RAW BLUE cells were cultured in a 96 well plate for 24 hours and then treated with 100ug/mL of sample. 24 hours after sample treatment, 20uL/well of 2mg/mL MTT solution was applied and cultured for 4 hours. After 4 hours of incubation, the medium was removed, DMSO was treated with 200 uL/well, and absorbance was measured at 540 nm after 1 hour of reaction.

At least three repeated experiments were performed, and the independent experimental results obtained from these were averaged and expressed as a percentage compared to the viability of control cells.

As a result, it was found that the cell viability was over 90% and that the sample processing concentration of 100ug/mL did not affect cytotoxicity (Figures 2 and 3).

Example 2: Establishment of colitis animal model and composition of experimental group

Based on the results of Example 1, the cytotoxicity was lowest in the mixed extract of Hongpalsac and Rehmannia glutinosa, especially when the mixing ratio of Hongpalsac and Rehmannia glutinosa was 3: 1, compared to the single extracts (extract of Red Palsy and Rehmannia glutinosa extract). After confirmation, the following experiments were conducted based on this.

2-1: Colitis animal model

The experimental animal used was a 5-week-old female ICR mouse purchased from Orient Bio Co., Ltd. (Korea). They were used in experiments after being adapted to the laboratory breeding environment for one week. The experimental animal center breeding conditions were 50 - 55% humidity, temperature 24 ± 1°C, and light/dark cycle: 12 hours. The experiment was conducted with the approval of the Animal Experiment Ethics Committee of Jeju National University (approval number: 2020-0030).

2-2: Experimental group

The colitis animal model of Example 2-1 was treated every day with drinking water containing HSE (Rhododendron chinensis + Rehmannia glutinosa + ripe Rehmannia glutinosa) and HSB (Rich chinensis + ripe Rehmannia glutinosa + citrus biogel) at a concentration of 1 mg/mL. Treatment was performed for 4 weeks, and changes were confirmed after 4 weeks. As a positive control group, mice treated with DSS (Dextran Sulfate Sodium) and only drinking water for 4 weeks were used.

Example 3: Experiment on the effect of mixed extract on DSS-induced enteritis model

3-1: Disease activity index (DAI)

After the start of the experiment, the weight of the mice in each group was measured every day, calculated as a weight loss rate, and the condition of the stool and the presence or absence of bloody stool were checked. For the above three indicators, the disease activity index (DAI; Elkatary et al. BJPR, 7(2): 140-151, 2015) scores were calculated according to the standards in Table 1 below.

Scoring criteria for calculating DAI Score weight (%) stool hardness Bleeding/occult blood reaction 0 no change hard stool normal One 1-5% reduction 1-2 loose stools anal swelling 2 6-10% reduction Soft stools <25% very mild bleeding,
1 to 5 bloody stools 3 10-15% reduction Soft stools 25-75% Severe bleeding, 5 or more bloody stools 4 >15% reduction diarrhea heavy bleeding

The sum calculated based on the score calculation criteria in Table 1 above is shown in Figure 4 as the disease activity index (DAI).

As a result of checking the DAI index after 2 weeks, it was confirmed that the DAI index, which had increased due to DSS treatment, decreased, and the DAI index was similar in the two treatment groups, HSE (Red Rehmannia glutinosa + Rehmannia glutinosa + Ripe Rehmannia glutinosa + Ripe Rehmannia glutinosa + Tangerine biogel). It was confirmed that the score was indicated.

3-2: Change in the length of the large intestine

As a result of measuring the entire length of the large intestine excluding the cecum, it was confirmed that the length of the large intestine in the DSS-administered group was shorter compared to the normal group, but after HSE or HSB treatment, the length of the large intestine was restored to that of the normal group (Figures 5a and 5a 5b). In particular, when comparing the two treatment groups (HSE or HSB), it was confirmed that the colon length of the HSB treatment group containing citrus biogel was recovered similar to that of the normal group.

Through this, it can be seen that tangerine (Hongpalsak) and/or Rehmannia glutinosa extracts have excellent effects on improving or treating enteritis, and are especially effective in improving enteritis when citrus biogel is included.

Example 4: Histopathological observation

Intestinal tissues obtained by sacrificing experimental animals were fixed with 10% neutralized formalin. The fixed intestinal specimen was infiltrated with paraffin, solidified, embedded, and then cut into 4-6 um sections. The cut sections were stained twice with hematoxylin and eosin dye and observed using an optical microscope.

As a result, compared to the normal group, the DSS-induced enteritis-induced group showed damage to the mucosal epithelium, loss of crypts, and increased infiltration of inflammatory cells and fibrosis, but it was confirmed that this trend was somewhat alleviated by HSE or HSB treatment (Figure 6 ). In particular, it can be seen in Figure 6 that the structure of D(HSB) containing citrus biogel is more relaxed.

Example 5: Short-chain fatty acid analysis

In the intestines, short-chain fatty acids act as nutrients for intestinal epithelial cells, affecting cell proliferation, differentiation, and gene expression, and acidifying intestinal pH. In addition, the intestinal environment changed due to the increase in short-chain fatty acids is known to cause decreased solubility of bile acids and decreased ammonia absorption, and changes intestinal minerals into a form that is easier to absorb. In particular, short-chain fatty acids are known to help improve colon health by promoting the regeneration of damaged colon.

Accordingly, the present inventors sought to determine whether short-chain fatty acids were increased even when the extracts (HSE and HSB) of the present invention were treated.

Specifically, short-chain fatty acids propionic acid and butyric acid were analyzed. As a result of the analysis, it was confirmed that when inflammatory bowel disease was induced with DSS, fatty acid production was reduced compared to the untreated group, and short-chain fatty acids increased when treated with HSE and HSB (Figure 7). In particular, the analysis results confirmed that more short-chain fatty acids were produced in the HSE-treated group. In addition, in the HSE group, it was confirmed that the distribution of microorganisms related to short-chain fatty acid production increased (Figure 8).

Example 6: Increase in fatty acid-related microorganisms

Acetatifactor or Alistipes is a beneficial bacteria that decreases in people suffering from intestinal diseases such as ulcerative colitis and increases in healthy people, and is known to produce short-chain fatty acids (SCFA), which are beneficial substances in the intestines (Jian Sun et al, 2020. International journal of biological macromolecules)

Accordingly, the present inventors sought to determine whether microorganisms producing short-chain fatty acids increased when the extracts (HSE and HSB) of the present invention were treated.

As a result, in this study, the microorganisms decreased significantly in the DSS group and tended to gradually recover in group B (HSB treatment) rather than group A (HSE treatment) (Figures 9a and 9b).

Example 7: Analysis of microbial diversity and homogeneity

It is known that when inflammatory colitis is induced by DSS treatment, the diversity and homogeneity of microbiota are reduced. Accordingly, the present inventors sought to determine whether the diversity and homogeneity of microbiota were increased even when treated with the extracts of the present invention (HSE and HSB).

For microbiota analysis, the nucleotide sequence of the 16S rRNA amplicon in experimental mouse feces was analyzed. For base sequence analysis, a wireless DNA quality check is required, so the amount of sample DNA was checked using the picogreen method using Victor 3 fluorometry, and then the quality of the DNA was checked through gel electrophoresis to finally decide whether to produce a library. . Once the DNA status is confirmed, DNA library production and quality check are performed. After producing the library and performing PCR, the size of the enrichment fragments was verified, the amount of the library was checked using qPCR according to the Illumina qPCR quantification protocol guide, and the final sample to be sequenced was determined. Base sequence analysis was performed using the MiSeq platform, which is widely used in intestinal microbiome research and produces genome data of 300 basepair size. The v3-v4 region of 16S rRNA amplified in the previous process was targeted. Next, bioinformatics analysis is performed, and the overall analysis pipeline uses the latest version of the QIIME2 program, which is most commonly used in recent research. A quality check was performed on the produced genome data, and low quality parts were trimmed and merged. A feature table was created using the OTU (Operational Taxonomic Unit) method and filtered out archae and Cyanobacteria that were unrelated to microbiome analysis. Using the NCBI RefSeq database that follows the latest nomenclature, we determined which bacterium each feature was, calculated the diversity index by referring to the final table, and performed statistical analysis.

As a result, it was confirmed that the diversity and homogeneity of microbiota increased in both the HSE and HSB treatment groups compared to the DSS treatment group (FIGS. 10a and 10b). Specifically, the diversity and homogeneity of microbiota were confirmed to be significantly reduced in the DSS group compared to the normal group, but the diversity and homogeneity of microbiota were confirmed to increase in the HAE and HSB treatment groups, similar to the normal group.

Example 8: Inflammatory cytokine analysis

To determine whether the extracts of the present invention (HSE and HSB) had an effect on improving inflammatory bowel disease induced by DSS, the production of inflammatory cytokines TNF-α and IL-6 was measured in serum or colon.

To measure inflammatory cytokine inhibitory activity, RAW BLUE cells were distributed in a 24 well plate at 1.5Х10 ⁵ cells/mL, cultured for 18 hours, and then treated with 1 μg/mL of LPS to induce cytokine production. Cells containing cytokines were treated with samples at different concentrations, cultured for 24 hours, and the supernatant was used to measure the inhibitory activity of cytokine production. Mouse TNF-α (Invitrogen Corp., Carlsbad, California, USA) and IL-6 (Invitrogen Corp., Carlsbad, California, USA) were measured using ELISA kits.

As a result, it was confirmed that in the DSS-treated group, the production of inflammatory cytokines rapidly increased compared to the control group, the untreated group (normal control group), and that the amount produced decreased upon sample intake (FIGS. 11a and 11b). This appears to help improve inflammatory bowel disease by reducing inflammation and reducing the production of cytokines (TNF-α, IL-6) due to the ingested samples. In particular, it was confirmed that the production of inflammatory cytokines was significantly reduced in the HSB-treated group containing citrus biogel.

This suggests that treatment with HSE and HSB may have a therapeutic effect on inflammatory diseases by reducing the secretion of inflammatory cytokines directly involved in the inflammatory response.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing its technical idea or essential features. In this regard, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed as including the meaning and scope of the patent claims described below rather than the detailed description above, and all changes or modified forms derived from the equivalent concept thereof are included in the scope of the present invention.

Claims (13)

A pharmaceutical composition for the prevention or treatment of inflammatory bowel disease, comprising extracts of Red Pale Root, Rehmannia glutinosa, and citrus biogel as active ingredients.
According to paragraph 1,
A pharmaceutical composition wherein the extract is extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.
According to paragraph 1,
The extract is a pharmaceutical composition comprising Hongpalsak and Rehmannia glutinosa in a weight ratio (w/w) of 1 to 3: 1.
delete According to paragraph 1,
A pharmaceutical composition comprising 5 to 15% by weight of the citrus biogel based on the total weight of the composition.
According to paragraph 1,
The inflammatory bowel disease is any one or more selected from the group consisting of enteritis, ulcerative colitis, Crohn's disease, intestinal Bechet's disease, bleeding rectal ulcer, and ileal pouchitis, Pharmaceutical composition.
The composition of claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient, or diluent.
A food composition for preventing or ameliorating inflammatory bowel disease, comprising as active ingredients extracts of Red Pale Root, Rehmannia glutinosa, and citrus biogel.
A feed composition for preventing or ameliorating inflammatory bowel disease, comprising extracts of Red Palsy, Rehmannia glutinosa, and citrus biogel as active ingredients.
A quasi-drug composition for preventing or improving inflammatory bowel disease, comprising extracts of Red Pale Root, Rehmannia glutinosa and citrus biogel as active ingredients.
A cosmetic composition for alleviating or improving inflammatory bowel disease, comprising extracts of Red Pale Root, Rehmannia glutinosa, and citrus biogel as active ingredients.
delete According to any one of claims 8 to 11,
A composition comprising 5 to 15% by weight of the citrus biogel based on the total weight of the composition.