KR102590321B1 - Immunologic function-improwing composition comprising cirsium japonicum extract as effective component - Google Patents

Abstract

The present invention relates to a composition for improving immune function containing thistle extract as an active ingredient, which increases the production of nitric oxide and the expression of cytokines related to immune function, The aim is to provide a composition for improving immune function that shows an effect of improving immune function and does not cause side effects even when the dosage and period of administration are increased.

Description

Composition for improving immune function containing thistle extract as an active ingredient {IMMUNOLOGIC FUNCTION-IMPROWING COMPOSITION COMPRISING CIRSIUM JAPONICUM EXTRACT AS EFFECTIVE COMPONENT}

The present invention relates to a composition for improving immune function containing thistle extract as an active ingredient. More specifically, thistle extract is included as an active ingredient to increase the production of nitric oxide and the expression of cytokines related to immune function, showing the effect of improving immune function, even if the dosage and period of administration are increased. It relates to a composition for improving immune function that can effectively prevent or treat immune-related diseases without problems with side effects.

The human body has the characteristic of trying to maintain homeostasis, and this also plays a role in the immune system. Immunity refers to a series of biological defense reactions that occur to exclude substances other than the body's own components from disrupting the body's homeostasis or threatening itself. It protects the body from harmful substances entering through the skin, digestive tract, respiratory tract, etc. It can be said to be a defense system that acts as a phagocytosis to remove external irritants or maintains homeostasis by reducing serious inflammation in wounds. All immune reactions, including the inflammatory response activated due to the introduction of pathogens into the body, return to normal after the pathogens are removed, but if there is difficulty in immune control, the immune response occurs significantly higher or lower than in normal people. do. A state in which the immune function is deficient or reduced is called immunodeficiency. In this state, the immune response is not properly activated and the body cannot respond properly to foreign substances in the body, resulting in infection. Conversely, an immune hypersensitivity reaction is when some immune responses overreact, creating an imbalance in the immune system, resulting in an allergic reaction.

When the immune system is not regulated normally and becomes imbalanced, immune control is impaired due to causes such as imbalance of cytokines in the body, imbalance of proliferation of T/B cells, and depletion of antioxidant nutrients, and the secretion of inflammatory substances increases, causing damage to cells and tissues. The inflammation may worsen due to the inability to cope with the influx of pathogens. Therefore, the body's immune response must be balanced and regulated normally to maintain health, so the ability to regulate immunity is important in disease prevention and treatment.

The spleen is involved in immune regulation and secretes cytokines such as interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-5, IL-6, and PGE2. These cytokines act at the execution stage of the immune response and also play an important role in the signal transmission process between immune cells and the inflammatory system.

Immune diseases are diseases in which components of the mammalian immune system cause, mediate, or otherwise contribute to pathological conditions in mammals, and inflammatory disorders in particular are one of the most important health problems worldwide. Inflammation is generally a localized protective response of body tissues against host invasion by foreign substances or harmful stimuli. Causes of inflammation include infectious causes such as bacteria, viruses, and parasites; Physical causes such as burns or irradiation; Chemical agents such as toxins, drugs or industrial agents; It may be a condition associated with an immune response, such as allergies and autoimmune reactions, or oxidative stress.

Inflammation is characterized by pain, redness, swelling, heat, and ultimate loss of function of the affected area. These symptoms are the result of a complex series of interactions between cells of the immune system. The cellular response results in the creation of an interactive network of several groups of inflammatory mediators: proteins (e.g. cytokines, enzymes (e.g. proteases, peroxidases), major basic proteins, and adhesion molecules (ICAMs). VCAM), lipid mediators (e.g. eicosanoids, prostaglandins, leukotrienes, platelet activating factor (PAF)), reactive oxygen species (e.g. hydroperoxide, superoxide anion O2-, nitric oxide (NO) ), etc.). However, many of these mediators of inflammation are also regulators of normal cellular activity. Thus, a deficiency in the inflammatory response leads to uncontrolled damage (i.e. infection) to the host, and thus, in part, to chronic inflammation. Inflammatory diseases are caused by excessive production of several of the above-mentioned mediators.

Additionally, autoimmune disease, one of the immune diseases, is characterized by the immune system attacking its own organs and causing a spontaneous response. These reactions are due to the recognition of auto-antigens by T lymphocytes, which triggers humoral (autoantigen production) and cellular (increased cytotoxic activity of lymphocytes and macrophages) immune responses. Autoimmune diseases include: rheumatic diseases, psoriasis, systemic dermatomyositis, multiple sclerosis, lupus erythematosus, or aggravated immune response due to antigens, such as asthma, drug or food allergy, etc. These are all limited and chronic diseases, and in some cases, they are fatal, and there is currently no effective treatment method to treat these diseases. Therefore, any drug, medicine, or medium that can alleviate or relieve the disease during the progression of these diseases will be an important solution for the patient's health.

Many researchers have made intensive efforts to explore treatment methods for autoimmune diseases and find appropriate drugs and methods. Today, the treatment of autoimmune diseases is mainly immunosuppressive drugs, such as glucocorticoids and calcineurin inhibitors. It is based on the use of antiproliferative santimetabolites and antiproliferative agents. However, since such pharmacological therapy acts on various targets, it can reduce overall immune function. Otherwise, if such pharmacological therapy is used for a long period of time, various cytotoxic effects may become problematic, suppressing the immune system in a non-specific manner, exposing the patient to the risk of infectious diseases and immune-related diseases. Calcineurin and glucocorticoids present additional problems due to their nephrotoxic and diabetic properties, so their use is limited in cases of some clinical conditions (e.g. renal dysfunction, diabetes, etc.).

Therefore, there is a need to develop new therapeutic agents that can treat immune-related diseases such as autoimmune diseases, inflammatory diseases, transplant rejection, etc., and have excellent therapeutic effects without side effects.

KR 10-2009-0001190 B1

The purpose of the present invention is to provide a composition for improving immune function that includes milk thistle extract as an active ingredient to increase the production of nitric oxide and the expression of cytokines related to immune function, thereby improving immune function. .

Another object of the present invention relates to a composition for improving immune function that contains thistle extract as an active ingredient and does not cause side effects even when the dosage and period of administration are increased.

A composition for improving immune function according to an embodiment of the present invention includes Cirsium japonicum extract as an active ingredient.

The thistle extract is extracted using an extraction solvent selected from the group consisting of water, alcohols with 1 to 6 carbon atoms, and mixtures thereof.

The composition contains a fermented thistle extract fermented through fermentation microorganisms.

The fermentation microorganisms include Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium bifidum, Bifidobacterium breve, and It is selected from the group consisting of Lactobacillus acidophilus.

The composition includes purified water, methanol, ethanol, propanol, isopropanol, butanol, alcohols having 1 to 4 carbon atoms, acetone, ether, Thistle extract fractionated with a solvent selected from the group consisting of benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, and mixtures thereof. It includes.

A food composition according to another embodiment of the present invention is prepared including the composition for improving immune function.

A pharmaceutical composition according to another embodiment of the present invention is prepared including the composition for improving immune function.

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

In the present invention, 'improvement of immunity' means inducing an initial immune response or measurably increasing an existing immune response to an antigen.

In the present invention, 'prevention' refers to any action that suppresses or delays the progression of the disease or disease to which the term applies, or one or more symptoms of the disease or disease, by administering the composition of the present invention.

In the present invention, 'improvement' means any action in which the disease or disease to which the term applies, or one or more symptoms of the disease or disease, is improved or beneficially changed by administration of the composition of the present invention.

In the present invention, unless otherwise stated, 'treatment' refers to reversing, alleviating, inhibiting the progression of, or preventing the disease or disease to which the term applies, or one or more symptoms of the disease or disease. The term 'treatment' used herein refers to the act of treating when 'treating' is defined as above.

As used herein, the term “active ingredient” refers to an ingredient that exhibits the desired activity alone or can exhibit activity in combination with a carrier that is not active on its own.

The term 'extract' used in this specification has the meaning commonly used in the art as a crude extract, as described above, but in a broad sense also includes fractions obtained by additional fractionation of the extract. In other words, plant extracts include not only those obtained using the above-described extraction solvent, but also those obtained by additionally applying a purification process. For example, fractions obtained by passing the extract through an ultrafiltration membrane with a certain molecular weight cut-off value, separation by various chromatographs (designed for separation according to size, charge, hydrophobicity, or affinity), etc. Fractions obtained through various purification methods are also included in the plant extract of the present invention.

The extract used in the present invention can be prepared in powder form by additional processes such as reduced pressure distillation and freeze drying or spray drying.

A composition for improving immune function according to an embodiment of the present invention includes Cirsium japonicum extract as an active ingredient.

Thistle (Cirsium japonicum) is a perennial herb entirely covered with hair. It grows upright with some branches at the top, reaching a height of about 1m. There are large and small thorns on the edges of the leaves. Purple or red flowers bloom upward one at a time at the ends of branches in midsummer. Each flower contains hundreds of small, tube-shaped flowers. The fruit has hairs and, when mature, is blown by the wind and reproduces. The young leaves are used as herbs and the whole is dried and used medicinally. Similar plants include big thistle, fin thistle, and Korean thistle.

Thistle is called Daegye as a medicinal herb and is known to have the effect of hemostasis of blood, detoxification of blood, and anti-inflammatory. In particular, thistle is known to have excellent hemostatic properties. The main ingredient that exhibits hemostatic properties is pectolinarin, and in experiments using mice, it was reported that a dose-dependent reduction in bleeding time was observed. In addition, thistle promotes the proliferation of lymphocytes, increases the activity of NK cells, and has tumor-suppressing properties, so it is believed to have anti-cancer and immune-boosting effects. In addition, it is used to treat colds, whooping cough, enteritis, nephritis, hematemesis, and hematuria. It is known to be used when bloody stool or postpartum bleeding does not stop, or to treat symptoms such as enlargement and boils.

The thistle extract according to the present invention can be obtained by extraction and separation from nature using extraction and separation methods known in the art, and 'extract' as defined in the present invention is extracted from thistle using an appropriate solvent. For example, it may include a hot water extract, a polar solvent-soluble extract, or a non-polar solvent-soluble extract of thistle.

The thistle extract is extracted using an extraction solvent selected from the group consisting of water, alcohols with 1 to 6 carbon atoms, and mixtures thereof.

Specifically, to prepare the extract, there are steps of washing the natural product; Drying after washing; Grinding the natural product after drying; leaching the pulverized material using an organic solvent; Leaching and drying the sample; Leaching using water; And including the step of leaching, a natural extract can be obtained.

The natural extract extracted using the organic solvent may further include fractionation using the organic solvent.

The method of preparing the extract may be a conventional extraction method in the art, such as ultrasonic extraction, leaching, and reflux extraction. Specifically, it may be an extract obtained by extracting a natural product from which foreign substances have been removed by washing and drying with water, alcohol having 1 to 6 carbon atoms, or a mixed solvent thereof, and may be an extract obtained by sequentially applying the solvents to the sample.

The reflux extraction method is based on 100 mL of water and alcohol with 1 to 6 carbon atoms, 10 to 30 g of pulverized natural product, reflux time of 1 to 3 hours, and 50 to 100% of alcohol or water with 1 to 6 carbon atoms. More specifically, based on 100 mL of alcohol with 1 to 6 carbon atoms or 100 mL of water, 10 to 20 g of pulverized natural product, reflux time of 1 to 2 hours, and 70 to 90% of alcohol or water with 1 to 4 carbon atoms. .

The leaching method is carried out at 15 to 30°C for 24 to 72 hours, and water or 50 to 100% alcohol with 1 to 6 carbon atoms is used as the extraction solvent. More specifically, the extraction is performed at 20 to 25°C for 30 to 54 hours, and the extraction solvent is water or 70 to 80% alcohol with 1 to 6 carbon atoms.

The ultrasonic extraction method proceeds at 30 to 50°C for 0.5 to 2.5 hours, and the extraction solvent is water or 50 to 100% alcohol with 1 to 6 carbon atoms. Specifically, extraction is performed at 40 to 50°C for 1 to 2.5 hours, and the extraction solvent is water or 70 to 80% alcohol with 1 to 6 carbon atoms.

The extraction solvent can be used in an amount of 2 to 50 times, more specifically, 2 to 20 times the weight of the sample. For extraction, the sample may be left in the extraction solvent for 1 to 72 hours, more specifically, 24 to 48 hours.

After extraction, the extract can be fractionated by sequentially applying a new fractionation solvent. The fractionating solvent used during fractionation is at least one selected from the group consisting of water, hexane, butanol, ethyl acetic acid, ethyl acetate, methylene chloride, and mixtures thereof, and is preferably ethyl acetate or methylene chloride.

After obtaining the extract or fraction, additional methods such as concentration or freeze-drying can be used.

The composition contains a fermented thistle extract fermented through fermentation microorganisms.

When using the fermented thistle extract, it has the advantage of being able to be used stably at a relatively high concentration due to its low cytotoxicity, and the preference that falls due to the off-flavor unique to the natural product can be increased through fermentation.

The fermentation microorganisms include Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium bifidum, Bifidobacterium breve, and It is selected from the group consisting of Lactobacillus acidophilus.

Microorganisms used in the fermentation include Lactobacillus salivarius, Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus rhamnosus, and Lactobacillus plantarum ( Lactobacillus plantarum), Lactobacillus helveticus, Lactobacillus fermentum, Lactobacillus paracasei, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus, lactobacilli, such as Lactobacillus reuteri, Lactobacillus buchneri, Lactobacillus gasseri, Lactobacillus johonsonii, Lactobacillus kefir, etc. Lactococcus lactis, Lactococcus plantarum, Lactococcus raffinolactis, Enterococcus faecalis, Enterococcus faecium, Streptococcus Lactobacillus coxes such as Streptococcus thermophilus, Leuconostoc lactis, Leuconostoc mesenteroides and Bifidobacterium animals, Bifidobacterium bifidium (Bifidobacterium bifidum), Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium pseudolongum, Bifidobacterium It may include Bifidobacteria such as Bifidobacterium themophilum and Bifidobacterium adolescentis, and more preferably Lactobacillus casei and Lactobacillus rhamnosus. rhamnosus), Bifidobacterium bifidum, Bifidobacterium breve, and Lactobacillus acidophilus. You can.

The composition includes purified water, methanol, ethanol, propanol, isopropanol, butanol, alcohols having 1 to 4 carbon atoms, acetone, ether, Fermented thistle extract fractionated with a solvent selected from the group consisting of benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, and mixtures thereof. It includes.

As an appropriate solvent for extracting the extract from thistle, any solvent acceptable in the art may be used, and water or an organic solvent may be used. For example, alcohols with 1 to 6 carbon atoms including purified water, methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, etc. , various solvents such as benzene, chloroform, ethyl acetate, methylene chloride, hexane, and cyclohexane can be used individually or in combination. It is not limited.

As a method, any one of the following methods can be selected and used: hot water extraction, cold needle extraction, reflux cooling extraction, solvent extraction, steam distillation, ultrasonic extraction, elution, and compression. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation to the method for producing the thistle extract of the present invention, and any known method can be used.

Preferably, the thistle extract included in the composition of the present invention is prepared by preparing the ethanol extract extracted by the above-described solvent extraction method into a powder state by additional processes such as reduced pressure distillation and freeze-drying or spray-drying, and then further using the solvent. Obtaining purified fractions.

The composition further comprises an herbal extract selected from the group consisting of rosemary, lavender, thyme, sage, mint, carob and mixtures thereof.

Herbal plants are herbaceous plants whose flowers, seeds, stems, leaves, and roots are useful to humans. Since ancient times, they have played an important role as medicinal herbs for the treatment of pain relief and sedation, as well as for preservatives and insecticides. Today, they are used as chemical herbs for the human body. It is a natural material that is receiving more and more attention due to its toxicity, concerns about environmental pollution, and its excellent efficacy.

Rosemary is a perennial evergreen low tree of the Lamiaceae family and usually grows to over 1 meter. The narrow, thin, needle-shaped leaves are tough like leather, are shiny, have a unique strong aroma, and are dark green. It has a sterilizing and disinfecting effect, so it is used to preserve meat and is often used in hospitals or when infectious diseases are prevalent. Rosemary helps improve memory, restores vitality when showing signs of lethargy, and eliminates headaches and migraines. It also has the effect of suppressing obesity, regenerates the skin, revitalizes sagging tissues, suppresses the occurrence of edema and dandruff, and promotes hair growth. Rosemary is used in cooking, for facial cleansing, and to raise blood pressure in cases of low blood pressure.

Lavender is the best-known variety of herbs. It is popular as an ornamental herb because its aromatic oil content makes the leaves and flower surfaces appear shiny, and its splendor shines even more during the flowering season. High-quality lavender is used for many purposes. In particular, the entire plant, including flowers, leaves, and stems, has a scent, so it is suitable for ornamental purposes as well as potpourri and various beauty products. For dried flowers, cut the purple buds before flowering in the morning and hang them in a well-ventilated shade to dry. Ornamental plants are cut down after the flowers fade, leaving about 2 nodes. Toner made with lavender essential oil is effective for rough skin because it has a cleansing effect that relieves skin tension and regenerates it cleanly and moisturized. Also, the scent of lavender has a calming effect, so it was put in a pillow and used for facial soothing. Drinking tea made with lavender has a sedative effect, relieves pain and headaches, and improves mood, helping you sleep well.

Thym is an aromatic plant belonging to the Lamiaceae family and has small grayish green leaves. There are two main varieties. One is the southern French variety (meridionale, also called serpolet). wild thyme) and the other is the cultivar (Thymus vulgaris), also called winter thyme (thym dhiver), German thyme (thym allemand), or farigoule, which is taller, has wider leaves, and a more bitter taste. . Lemon thyme (thym citron, T. citriodorus) is also widely used to give a refreshing flavor to various dishes. Thyme has a very pleasant scent and contains the essential oil thymol, which has antiseptic properties. Thyme, one of the basic cooking herbs, can be used fresh or dried and used alone or used to make a bouquet garni.

Sage has been used as a panacea since Greek and Roman times. It is used in a variety of ways, including for ornamental purposes, spice collection, medicine, cooking, and dyeing. It has tonic, soothing, digestive, and sterilizing effects. It is very essential in cooking because it neutralizes the fat in meat or fish and removes odors. Sage has many uses, but large amounts should not be used at once. Sage tea is effective in clearing the mood, calming excitement, and preventing oral inflammation, bleeding gums, and bad breath, but avoid drinking it continuously as it is very effective. In addition, sage promotes digestion, has a detoxifying effect, is used as a placebo, and as a bathing agent, it has a fatigue recovery effect, expectorant, carminative, diuretic, anthelmintic, and antiseptic, and is used to treat cold symptoms. It is very effective in treating menopausal disorders.

Mint is native to Europe, and along with lemon balm, it can be said to be a representative herb of Milwon plants. It is a perennial plant of the Lamiaceae family and is one of the strongest growing herbs. Just brushing the leaves has a strong minty scent that gives a refreshing and refreshing feeling. When storing, cut off the bottom of the stem in June or July when the scent is strongest and dry it in the shade. Mint-type herbs, which are characterized by a refreshing sensation, are especially used in making sauces for meat dishes, and their refreshing taste and aroma are used to flavor vinegars sprinkled on vegetable or fruit salads, drinks made with carbonated water, and cocktails. Although there are some differences depending on the type, mints are used as raw materials for sterilization, promoting digestion, acting on the stomach, oral deodorants, toothpaste, and placebo. It has medicinal properties for dermatitis and itching, and when placed in the bathroom, the peppermint scent lasts for a long time instead of the bad smell, and it also has a strong sterilizing effect. Green mint called spearmint is often used in cooking along with peppermint. It has a sweeter scent than peppermint and is very good for the skin.

Carob {Ceratonia siliqua (L.) Taub.} is an evergreen broad-leaved shrub that grows to about 10 m in height and is drought tolerant. The leaves contain resin. Small, dark greenish-brown flowers bloom in the fall, and long green pods hang from them. These pods turn chocolate-colored when ripe, and contain small, shiny, hard beans inside. Carob gum, which softens the skin, is popular as a skin pack and is also used to treat diarrhea.

Preferably, the composition of the present invention contains 40 to 60 parts by weight of rosemary extract, 40 to 60 parts by weight of lavender extract, 40 to 60 parts by weight of thyme extract, 40 to 60 parts by weight of sage extract, and 40 parts by weight of mint extract, based on 100 parts by weight of thistle extract. It may contain 40 to 60 parts by weight of carob extract and 40 to 60 parts by weight of carob extract.

When used as a complex extract within the above range, the complex action between each component can increase the production of nitric oxide and increase the expression of immune function-related cytokines, so when using only thistle extract, In comparison, it can be provided as a composition for improving immune function with excellent immune function improvement effect.

In addition, it makes it possible to provide excellent food compositions with no side effects as ingredients derived from natural products.

Preferably, the composition further includes a natural extract selected from the group consisting of Aster fastigiatus Fisch. extract, Vicia pseudoorobus Fisch. & C.A.Mey., and mixtures thereof.

The Aster fastigiatus Fisch. is a perennial herb and is distributed in Korea, China, Japan, and Russia. In Korea, it is distributed throughout the country and grows to a height of about 30 to 100 cm. The stem is 30-100cm high, stands upright, has vertical ridges, branches spread out in a corymb shape from the upper part, and coarse hairs grow densely. Flowers bloom from August to October, are 7-9mm in diameter, hang on corymbs at the end of the main stem, and the peduncles are 3-8mm long. The involucre is barrel-shaped, 4mm long and 5mm wide. The bracts are arranged in 4 rows, have oblanceolate blunt heads, are hairy, and the inner piece is 1.5mm long. The snowflakes are one row, white, and the corolla is 5-6.5mm long and 1mm wide. The leaves that first emerge remain as they are until the flowers bloom, are linear-lanceolate, have narrow ends, are 5-12cm long, and are 4-15mm wide. The bottom narrows to become a petiole, and the back side is whitish, with glandular dots and ventral hairs. It has sparse sawtooth edges, is often curled backwards, and has short hairs on the upper edge. The cauline leaves gradually become smaller as they go upward and are linear-lanceolate or linear. The back side is densely covered with compound hairs and has glandular dots. The leaves of the inflorescence are 2-3mm long.

The big-backed rake (Vicia pseudoorobus Fisch. & C.A.Mey.) is a perennial vine whose stems are 80 to 150 cm long and have some or no hairs. The leaves are pinnately compound leaves composed of 2 to 5 pairs of leaflets, and the tendrils at the ends are not split or divided. The leaflets are ovate, acute or obtuse, 3 to 5 cm long and 15 to 30 mm wide. When they dry, they turn yellow-brown, the veins on the back protrude, and the lower leaves are green and sharply split. Flowers emerge from the leaf axils, are 4 to 8 cm long, have long peduncles, are slanted from the center to one side, and many flowers grow in racemes. The flowers bloom in September, are 13-15mm long, are purple, and the sepals are low triangular. The leaves are pinnate compound leaves composed of 2-5 pairs of leaflets, and the tendrils at the ends are not split or split. The leaflets are ovate, acute or obtuse, 3-5cm long and 15-30mm wide. They turn yellow-brown when dry, the veins on the back protrude, and the leaflets are green and sharply split.

When the composition further contains, in addition to thistle extract and herbal extract, a natural extract selected from the group consisting of Onggut herb extract, Great-backed Rake extract, and mixtures thereof, as it is composed of a complex extract, only thistle extract is used as a mixture. Compared to the case of using only thistle extract, it can increase the production of nitric oxide and increase the expression of immune function-related cytokines, and has an excellent immune function improvement effect compared to the case of using only thistle extract. makes it possible to provide

Preferably, the composition of the present invention contains 40 to 60 parts by weight of rosemary extract, 40 to 60 parts by weight of lavender extract, 40 to 60 parts by weight of thyme extract, 40 to 60 parts by weight of sage extract, and 40 parts by weight of mint extract, based on 100 parts by weight of thistle extract. It may include 60 to 60 parts by weight of carob extract, 40 to 60 parts by weight of carob extract, 5 to 10 parts by weight of Onggut sprout extract, 5 to 10 parts by weight of Great-backed Rake extract, and 5 to 10 parts by weight of Hairy Primrose extract.

When used as a complex extract within the above range, the immune function improvement effect is further increased due to the complex action between each component.

A food composition according to another embodiment of the present invention is prepared including the composition for improving immune function.

The composition for improving immune function can be added as is or used together with other foods or food ingredients, 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 (prevention, health, or therapeutic treatment). Generally, when producing a food or beverage, the composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw materials. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. .

There are no special restrictions on the types of foods above. Examples of foods to which the composition can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, These include alcoholic beverages and vitamin complexes, and include all health foods in the conventional sense.

The health drink composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients like conventional drinks. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As a sweetener, natural sweeteners such as thaumarin and stevia extract or synthetic sweeteners such as saccharin and aspartame can be used.

In addition to the above, the health drink composition of the present invention contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, It may contain carbonating agents used in alcohol and carbonated beverages. In addition, the health drink composition of the present invention may contain pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients can be used independently or in combination.

A pharmaceutical composition according to another embodiment of the present invention is prepared including the composition for improving immune function.

The composition for improving immune function of the present invention may contain appropriate carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions, and the pharmaceutical dosage form of the composition may be used alone or in combination with other pharmaceutically active compounds. Can be used in suitable sets.

The composition for improving immune function according to the present invention is formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions according to conventional methods. It can be used interchangeably. Carriers, excipients, and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, and methyl. Various compounds or mixtures may be included, including 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. These solid preparations contain at least one excipient such as starch, calcium carbonate, or sucrose. ) or prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, oral solutions, emulsions, syrups, etc. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. . 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 preferred dosage of the composition of the present invention varies depending on the patient's condition and weight, degree of disease, drug form, administration route and period, but can be appropriately selected by a person skilled in the art. However, for desirable effects, the composition of the present invention is preferably administered at 0.0001 to 100 mg/kg per day, preferably at 0.001 to 100 mg/kg. Administration may be administered once a day, or may be administered several times. The above dosage does not limit the scope of the present invention in any way.

The composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans through various routes. All modes of administration are contemplated, for example, oral, rectal or by intravenous, intramuscular, subcutaneous, intrathecal or intracerebroventricular injection.

According to the composition for improving immune function containing thistle extract as an active ingredient of the present invention, it increases the production of nitric oxide (Nitric Oxide) and increases the expression of immune function-related cytokines, thereby improving immune function. By showing an improvement effect, it is possible to provide a composition for improving immune function that does not cause side effects even when the dosage and period of administration are increased.

Figure 1 is a graph measuring cell survival rate by a composition for improving immune function according to an embodiment of the present invention.
Figure 2 is a graph confirming the ability to induce nitric oxide (NO) production by a composition for improving immune function according to an embodiment of the present invention.
Figure 3 is a graph showing cytokine expression by a composition for improving immune function according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement it. However, the present invention may be implemented in many different forms and is not limited to the embodiments described herein.

[Preparation Example 1: Preparation of thistle extract]

1. Preparation of thistle extract

First, the thistle was washed, dried, and crushed. The pulverized product was mixed with ethanol, maintained at 98 to 100° C. for 2 hours, cooled, and filtered with Whatman filter paper to obtain the filtrate, which was prepared as thistle ethanol extract (JE).

2. Preparation of fermented thistle extract

Among the microorganisms used for fermentation of the thistle ethanol extract (JE) 10% (w/w), Streptococcus thermophilus was mixed with D.W 75% (w/w), Jinji powdered milk 15% (w/w) Fermented thistle extract (SJE) was prepared by inoculating the mixed sterilized medium and culturing (25°C, 24 hours).

3. Preparation of thistle fraction

The thistle ethanol extract (JE) was suspended in 2L of distilled water, fractionated with an equal amount of methylene chloride, and then concentrated under reduced pressure to obtain a thistle fraction (JME).

4. Preparation of fermented milk thistle fraction

The fermented thistle extract (SJE) was suspended in 2L of distilled water, fractionated with an equal amount of methylene chloride, and then concentrated under reduced pressure to obtain a fermented thistle fraction (SJME).

[Preparation Example 2: Preparation of complex extract]

1. Preparation of herbal extract

First, the rosemary was thoroughly washed under running water and then completely air-dried. Dried rosemary was ground in a blender and then made into powder. The powder sample was completely immersed in 50% ethanol as an extraction solvent at a ratio of 1:10 (w;v), and then extracted repeatedly three times for 3 hours while refluxing at 80°C. The extract was Whatman No. 2 Filtered with filter paper. The filtrate was concentrated under reduced pressure at 60°C to prepare rosemary extract (RH).

Lavender extract (LE), thyme extract (TE), sage extract (SE), mint extract (ME), and carob extract (KE) were prepared using the same method as the rosemary extract (RH).

3. Preparation of other extracts

Using the same method as the method for producing thistle ethanol extract (JE) prepared in Preparation Example 1, Onggut herb extract (AE) and Great-backed rake extract (VE) were prepared.

4. Preparation of complex extract

Thistle ethanol extract (JE), fermented thistle extract (SJE), thistle fraction (JME), fermented thistle fraction (SJME), rosemary extract (RH), lavender extract (LE), and sage extract (SE) prepared in Preparation Example 1. ), mint extract (ME), carob extract (KE), onggut extract (AE), and venom extract (VE) were mixed in the content ranges shown in Table 1 below to prepare a composite extract.

IM1 IM2 IM3 IM4 IM5 IM6 IM7 IM8 IM9 IM10 IM11 IM12 IM13 IM14 IM15 J.E. 100 - - - 100 100 100 100 100 100 100 100 - - - S.J.E. - 100 - - - - - - - - - 100 - - JME - - 100 - - - - - - - - - - 100 - SJME - - - 100 - - - - - - - - - - 100 R.E. - - - - 20 40 60 80 60 60 60 60 60 60 60 LE - - - - 20 40 60 80 60 60 60 60 60 60 60 S.E. - - - - 20 40 60 80 60 60 60 60 60 60 60 M.E. - - - - 20 40 60 80 60 60 60 60 60 60 60 KE - - - - 20 40 60 80 60 60 60 60 60 60 60 A.E. - - - - - - - - 3 5 10 13 10 10 10 VE - - - - - - - - 3 5 10 13 10 10 10

(Unit: parts by weight)

[Experimental Example 1: Measurement of cell viability]

cell culture

RAW 264.7 cells, a mouse macrophage cell line, were purchased from the Korea Cell Line Bank (KCLB) and used. The RAW 264.7 cells were cultured in an incubator controlled at 37°C and 5% CO ₂ using DMEM medium containing 10% FBS and 1% penicillin-streptomycin.

Cell viability measurements

RAW 264.7 cells were distributed in a ⁹⁶ -well plate at a concentration of 1 As a positive control, 1 ug/mL of lipopolysaccharide (LPS) was treated. After culturing for 24 hours, CCK-8 solution (10 mL/well) was added to each well and treated at 37°C for 2 hours to produce 450 mL. Absorbance was measured using an ELISA reader (TECAN, Mnnedorf, Switzerland) at nm wavelength.

The results are shown in Figure 1 below. As shown in Figure 1, the complex extracts IM1 to IM15 prepared in Preparation Example 2 showed a survival rate of more than 80% at all concentrations compared to the positive control, confirming that there was no cytotoxicity. .

[Experimental Example 2: Ability of thistle extract to induce nitric oxide (NO) production]

In order to confirm the immune enhancing effect of thistle extract (JE, SJE, JME and SJME) of the present invention, its effect on nitric oxide inducing ability was confirmed.

First, RAW264.7 cells (TIB-71, ATCC) were plated and stabilized in a 96-well cell culture plate at 1 × 10 ⁴ cells/well, and then diluted in DMEM medium with seaweed water extract (20 and 40 ㎍/㎖). and thistle extract (JE, SJE, JME, and SJME) at 150 ㎍/㎖, and as a positive control, 1 ㎍/㎖ LPS (Lipopolysaccharide, L4291, Sigma, St. Louis, MO, USA) was treated for 24 hours. Cultured. Add 50 ㎕ of cell culture supernatant and an equal amount of Griess reagent (G4410, Sigma, St. Louis, MO, USA) to a new 96-well plate, react at room temperature for 15 minutes, and then use a microplate reader (Molecular Devices, CA, USA), the absorbance was measured at 540 nm. The concentration of nitric oxide (NO) was calculated using a standard curve obtained using sodium nitrite (S2252, NaNO2, Sigma, St. Louis, MO, USA).

The results are shown in Figure 2 below. As a result of measuring the secretion amount of NO in RAW264.7 cells by thistle extract (JE, SJE, JME and SJME), the secretion of NO from fermented thistle extract (SJME) was concentration-dependent. It was confirmed that it was increasing.

The NO secretion ability of thistle extracts (JE, SJE, JME, and SJME) all showed a lower increase than that of the LPS-treated group, which was the positive control group.

[Experimental Example 3: Cytokine expression ability of thistle extract]

In order to confirm the immune-enhancing effect of thistle extract (JE, SJE, JME and SJME) of the present invention, its effect on cytokine expression was confirmed.

First, RAW264.7 cells (TIB-71, ATCC) were stabilized by dispensing at a concentration of 5 × 10 ⁶ cells/well in 60 mm cell culture dishes, and then 150 μg/well of thistle extract (JE, SJE, JME and SJME). ㎖ and LPS 1㎍/㎖ as a positive control for 4 hours, RNA was isolated using the RNeasy extraction kit (Qiagen, Gaithersburg, Maryland, USA) according to the manufacturer's experimental method. cDNA was synthesized from the isolated RNA using PrimeScript TM reverse transcriptase (TAKARA BIO INC., Shiga, Japan). To measure the expression of genes, real-time PCR was performed using the FastStart Universal SYBR Green Master (ROX) (Roche Diagnostics, Mannheim, Germany) using a PIKOREAL 96 (Thermo SCIENTIFIC, MA, USA) instrument. It was carried out using

The base sequence of each primer used in the analysis is GAPDH forward 5'-CAT GGC CTT CCG TGT TCC TA-3 (sequence list no. 1), reverse 5'-GCG GCA CGT CAG ATC CA-3' (sequence list no. 2) Sequence), IL-2 forward 5′-CCT GAG CAG GGA GAA TTA CA-3′ (sequence list no. 3), reverse 5′-TCC AGA ACA TGC CGC AGA-3′ (sequence list no. 4), IL -4 Forward 5'-AGA TGG ATG TGC CAA ACG TCC TCA-3' (Sequence Listing No. 5), Reverse 5'-AAT ATG CGA AGC ACC TTG GAA GCC-3' (Sequence Listing No. 6), IFN- γ Forward 5'-GGC CAT CAG CAA CAA CAT AAG CGT-3' (sequence list no. 7), reverse 5'-TGG GTT GTT GAC CTC AAA CTT GGC-3' (sequence list no. 8).

The results are shown in Figure 3 below. As a result of measuring the cytokine secretion ability in RAW264.7 cells by thistle extract (JE, SJE, JME, and SJME), the fermented thistle extract (SJME) showed IL-2, IL- It secretes excellent cytokines such as 4 and IFN-γ, and these cytokines play a role in promoting immune responses with other immune cells, confirming the immune-enhancing effect of thistle extract.

[Experimental Example 4: Immune function improvement effect of complex extract]

In order to confirm the effect of the composition for improving immune function of the present invention, the complex extracts IM5 to IM15 prepared in Preparation Example 2 were tested in the same manner as Experiments 1 to 3, and showed an immune function improvement effect. For comparison, the fermented milk thistle fraction (IM4, SJME) that showed excellent effects was set at an index of 1 and is shown in Table 2 below.

IM4, SJME IM5 IM6 IM7 IM8 IM9 IM10 IM11 IM12 IM13 IM14 IM15 Immune function improvement effect One 3 5 5 4 4 5 6 5 7 8 9

(Unit: index)

As shown in Table 2, in the case of the complex extract (IM15) containing fermented thistle fractions (IM4, SJME), thistle ethanol extract (JE), fermented thistle extract (SJE), and thistle fraction (JME) It was confirmed that compared to the complex extracts IM1 to IM14, it showed an equal or greater effect on improving immune function.

[Experimental Example 5: Evaluation of palatability]

A palatability evaluation was conducted on IM1 to IM15 of the present invention. After IM1 to IM15 were made into tea, they were provided to 20 adult men and women and asked to evaluate the taste and aroma.

Evaluation scores were requested from 1 to 10 points for each item, and the evaluation results were converted into average scores. The higher the number, the higher the preference for the index.

IM1 IM2 IM3 IM4 IM5 IM6 IM7 IM8 IM9 IM10 IM11 IM12 IM13 IM14 IM15 taste One 3 3 4 5 7 7 4 6 7 8 7 8 8 9 incense 2 4 3 4 5 5 6 5 7 7 7 6 8 9 9 Comprehensive preference (average) 1.5 3.5 3.0 4.0 5.0 6.0 6.5 4.5 6.5 7.0 7.5 6.5 8.0 8.5 9.0

(Unit: index)

Referring to Table 5 above, it was confirmed that preference for IM1 to IM4 decreased due to the unique taste and aroma of thistle. However, when using a mixture of other natural extracts of IM5 and IM12, it was confirmed that the preference was excellent.

In particular, in the case of IM15, a complex extract containing fermented milk thistle fractions, it was confirmed that it exhibited relatively excellent taste and aroma, confirming that it could be provided as a food composition with high preference.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims are also possible. falls within the scope of rights.

Claims (7)

Contains thistle (Cirsium japonicum) extract as an active ingredient,
For 100 parts by weight of thistle extract, 40 to 60 parts by weight of rosemary extract, 40 to 60 parts by weight of lavender extract, 40 to 60 parts by weight of thyme extract, 40 to 60 parts by weight of sage extract, 40 to 60 parts by weight of mint extract, carob Containing 40 to 60 parts by weight of extract, 5 to 10 parts by weight of Onggut sprout extract, and 5 to 10 parts by weight of Great-backed Rake extract.
Composition for improving immune function. According to clause 1,
The thistle extract is extracted using an extraction solvent selected from the group consisting of water, alcohols with 1 to 6 carbon atoms, and mixtures thereof.
Composition for improving immune function. According to clause 1,
The composition includes a fermented thistle extract fermented through fermentation microorganisms.
Composition for improving immune function. According to clause 3,
The fermentation microorganisms include Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium bifidum, Bifidobacterium breve, and Selected from the group consisting of Lactobacillus acidophilus
Composition for improving immune function. According to clause 1,
The composition includes purified water, methanol, ethanol, propanol, isopropanol, butanol, alcohols having 1 to 4 carbon atoms, acetone, ether, Thistle fraction fractionated with a solvent selected from the group consisting of benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, and mixtures thereof. containing
Composition for improving immune function. Comprising the composition for improving immune function according to any one of claims 1 to 5.
Food composition. Comprising the composition for improving immune function according to any one of claims 1 to 5.
Pharmaceutical composition for improving immune function.