KR20190129245A - Polysaccharide fraction isolated from Nelumbo nucifera leaf with immune-enhancing activity and method for producing the same - Google Patents

Abstract

The present invention relates to a polysaccharide fraction derived from a Nelumbo nucifera leaf having immune function enhancing activity and a manufacturing method thereof. The polysaccharide fraction, compared to a total polysaccharide fraction, contains: 45 to 70 wt% of neutral sugar; 20 to 45 wt% of uronic acid; 0.5 to 10 wt% of KDO-like substances; 3 to 20 wt% of protein; and 0.1 to 5 wt% of phenol, so that immune functions are improved and there is no toxicity, thereby being able to be consumed in a form of food. Therefore, it is possible to improve immune functions through life diet and lifestyle.

Description

Polysaccharide fraction derived from lotus leaf having immune function enhancing activity and preparation method thereof Polysaccharide fraction isolated from Nelumbo nucifera leaf with immune-enhancing activity and method for producing the same

The present invention relates to a lotus leaf-derived polysaccharide fraction and a method for preparing the same, which can be ingested in the form of food because it has excellent immune function and no toxicity, thereby improving immune function through diet and lifestyle in daily life. .

As the standard of living has risen and westernized eating has become more common, lifestyle diseases such as obesity, diabetes, hypertension, and heart disease due to overnutrition and lack of exercise are increasing. In addition, as the speed of entry into an aging society is increasing due to the extension of life expectancy, interest in health promotion is increasing.

Health promotion aims to increase resistance to disease, especially chronic degenerative or infectious diseases, ongoing stress, and daily activity. These measures include improving nutrition, strengthening immune function, proper exercise and rest, and mental health. Management over the entire lifestyle, including the continuation of activities, is necessary.

In particular, high immunity usually does not take the disease well, even if you get sick quickly recovery. Obviously, staying healthy and extending your life as you age. Therefore, immunity is directly related to human health and lifespan, and it is essential to manage immunity to maintain health and prolong life.

Immunity refers to a mechanism for physiologically recognizing and excluding physiologically foreign and endogenous foreign bodies in humans and animals. Here, the foreign material is called an antigen, and numerous kinds of white blood cells and proteins are involved in the process of removing it. Immunity is divided into innate immunity, which has been born since birth, and acquired immunity obtained by adapting to life.

Innate immunity is also called natural immunity or natural resistance. It reacts nonspecifically to the antigen and has no special memory effect. Innate immune systems include skin and mucous tissues that block the invasion of antigens, acid in the stomach, and complement in the blood. Cells include macrophages responsible for phagocytosis, polymorphonuclear leukocytes, and K cells that can kill infected cells. There is also a cytokine produced by macrophages. Indeed most infections are protected by this innate immunity.

Acquired immunity plays a role in reinforcing innate immunity and is also known as acquired immunity and adaptive immunuty. When an antigen invades the body, an immune response caused by lymphocytes occurs, and strong resistance to the antigen is remembered, and when the antigen is invaded again, the antigen reacts specifically to effectively remove the antigen. Lymphocytes are one of the white blood cells that are responsible for immune responses in vertebrates and mammals, accounting for about 25% of all white blood cells, and mature and deformed in the thymus to retain immune antibodies. Invasion is a cell that responds primarily to the body's immune response, preventing its antigen from reacting in the body.

There are two types of lymphocytes, which are classified into T cells responsible for cellular immunity and B cells responsible for humoral immunity. T cells are cells differentiated into lymphocytes by special internal environment and thymus fluid factor in thyroid epithelial cells, stimulate antibody production to B cells and directly attack antigens such as tumor cells. B cells, which are independent of the thymus, differentiate into lymphocytes in the bone marrow, and mature into antibody-producing cells upon antigen-stimulation and T cell-mediated stimulation to produce and secrete antibodies of IgM, IgG, IgA, and IgE.

Initial recognition of antigen infection is primarily performed by macrophage. In the process of macrophage phagocytosis, cytokines are produced that mediate the immune response.

Cytokines are glycoproteins that are used as signaling materials to control and stimulate the body's defenses and play important roles in immunity, infectious diseases, hematopoietic function, tissue repair, cell development and growth. Cytokines are called inflammatory cytokines because they function to localize immune cells by inducing inflammation at local sites. The most important immune functions are performed by a group of cytokines known as Interleukin-1, -6, -12, interferon gamma (IFN-γ) and tumor necrosis factor (TNF).

Interleukin (Interleukin) is a substance that plays an important role in the body's defenses by controlling the immune response by acting at various stages of the immune response, the kind is IL-1,6,12. Among them, IL-12 is a cytokine composed of p35 and p40 proteins, and it is an important immunological function that links innate and acquired immunity, and is a macrophage (macrophage) that is involved in the removal of endogenous antigens. ), Activates NK cells and T-cells.

Tumor necrosis factor (TNF) is a substance that necroses tumors secreted from T cells by the endotoxin, which is an endotoxin from the bacterial wall. It is the most important inflammatory factor and sometimes causes severe systemic inflammation. . However, from the immunological point of view, TNF aggregates neutrophils and monocytes, which are blood cells into the site of infection, and induces differentiation and activation of effector cells into killer cells. The function of the TNF is due to the ability to induce the production of cytokines having chemotaxis and to enhance the affinity of adhesion molecules (adhesionmolecules) between vascular endothelial cells and leukocytes.

As such, immune function is achieved by the action of a wide variety of cells such as macrophages, lymphocytes (T cells, B cells) and proteins such as cytokines such as interleukin and TNF.

Meanwhile, Nelumbo nucifera ( Sacred lotus) is a perennial aquatic plant belonging to the Nymphaeaceae family and is native to South Asia and North Australia. This plant is widely distributed in tropical and temperate eastern Asia including Korea and China, and in Korea and Japan. It is inhabited by flooded areas, lakes, wetlands, swamps, lakes and ponds within 2.5m of water. Sacred and cultivated a lot for ornamental purposes. It is known that cultivation of lead for edible purposes in Korea is about 50 years ago, and recently, farmers planting kites in ponds or paddy fields as profitable crops are increasing in large numbers.

The lotus is divided into lotus root of red, white or white lotus, lotus of lotus and lotus leaf. In other words, lotus has been widely used for food and medicinal purposes according to the parts of lotus leaf, lotus seed, lotus root, etc., and lotus has been used as a material for ornamental and tea. In Asia, they have eaten rice wrapped with leaves, and young leaves or stems are eaten as salads or pickles with other vegetables, and the roots are cooked variously as ingredients for various dishes. . In Korea, lotus root (underground stem) has been used as a side dish for a long time, and it is used as lotus tea and lotus leaf tea by drying flowers and leaves of white lotus, and it is known as tea for drinking diabetes and liver function. .

Lotus leaf contains alkaloids such as nelumboside, nuciferine, coclaurine, roemerine, etc. It has a calming and antipyretic effect, and is known to be effective in diabetes and hyperlipidemia. It is known to contain physiologically active ingredients such as flavonoids such as isoquercitrin, hyperoside, rutin and kaempferol. In addition, studies on components such as tartaric acid, citric acid, malic acid, succinic acid and tannin have been reported.

On the other hand, according to the results of the analysis of the general composition of the lotus leaf, the lotus leaf powder contains 0-2.98% of moisture, 0.93-1.0% of crude fat, 16.9-23.9% of crude protein, 8.1-9.3% of crude ash, and 63.8-65.2% of carbohydrate. (Lee et al., 2008; Park et al., 2009; Park et al. 2010). That is, the sugar component is a major component that is high in more than 60% of the dried lotus leaf. Nevertheless, studies on the efficacy of the sugar component of lotus leaf have reported that Zeng et al. (2016) showed antidiabetic activity of lotus leaf-derived selenium-polysaccharide in gestational diabetic rats. The research is insufficient.

Republic of Korea Patent Publication No. 10-2015-0000219 Republic of Korea Patent No. 10-1176435

It is an object of the present invention to provide a lotus leaf-derived polysaccharide fraction capable of improving immune function.

In addition, another object of the present invention to provide a method for producing the lotus leaf-derived polysaccharide fraction.

In addition, another object of the present invention to provide a pharmaceutical composition for the prevention or treatment of diseases caused by a decrease in immunity comprising the lotus leaf-derived polysaccharide fraction as an active ingredient.

In addition, another object of the present invention to provide a health food composition for enhancing immune function comprising the lotus leaf-derived polysaccharide fraction as an active ingredient.

In addition, another object of the present invention to provide a cosmetic composition for enhancing immune function comprising the lotus leaf-derived polysaccharide fraction as an active ingredient.

Lotus leaf-derived polysaccharide fraction of the present invention for achieving the above object is 45 to 70% by weight of neutral sugar, 20 to 45% by weight of acidic polysaccharide (uronic acid) and 0.5 to 10% by weight of KDO analogous substance May be included as a%.

The lotus leaf-derived polysaccharide fraction may further comprise 3 to 20% by weight of protein and 0.1 to 5% by weight of phenol.

The neutral polysaccharide may include arabose (arabinose), rhamnose (rhamnose), galactose (galactose), xylose, glucose (glucose), mannose and fucose (fucose).

The acidic polysaccharide may be composed of galacturonic acid and glucuronic acid.

The KDO-like substance may be composed of 3-deoxy-D-lyxo-2-heptulosaric acid (DHA) and 3-deoxy-D-manno-2-octulosonic acid (KDO).

The lotus leaf-derived polysaccharide fraction may be a polysaccharide having a molecular weight of 5 to 550 KDa containing 50% by weight or more.

The lotus leaf-derived polysaccharide fraction may have an immune function enhancing activity.

In addition, the method for producing a lotus leaf-derived polysaccharide fraction of the present invention for achieving the above another object comprises the steps of (A) extracting a mixture of lotus leaf powder and a solvent to obtain an extract; And (B) fractionating the extract by a solvent immersion or non-thermal treatment process.

In step (A), the solvent may be water, lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

Lotus leaf-derived polysaccharide fraction fractionated in step (B) is 45 to 70% by weight neutral sugar, 20 to 45% by weight acidic polysaccharide (uronic acid), 0.5 to 10% by weight KDO analogous substance It may include.

The lotus leaf-derived polysaccharide fraction may further comprise 3 to 20% by weight of protein and 0.1 to 5% by weight of phenol.

In addition, the pharmaceutical composition for the prevention or treatment of diseases caused by lowering the immunity of the present invention for achieving the above another object may include the lotus leaf-derived polysaccharide fraction as an active ingredient.

The disease caused by the reduced immune function may be selected from the group consisting of infectious diseases, inflammatory diseases, allergic diseases or chronic fatigue.

In addition, the health food composition for enhancing the immune function of the present invention for achieving the above another object may include the lotus leaf-derived polysaccharide fraction as an active ingredient.

In addition, the cosmetic composition for enhancing the immune function of the present invention for achieving the above another object may include the lotus leaf-derived polysaccharide fraction as an active ingredient.

Lotus leaf-derived polysaccharide fraction of the present invention is very excellent in improving the immune function, there is no toxicity can improve the immune function through the diet and lifestyle of daily life in the long term.

 In addition, the lotus leaf-derived polysaccharide fraction of the present invention is effective in the production of competitive foods, immune-enhancing cosmetics and immune-enhancing drugs.

Figure 1a is a molecular weight graph for the hydrothermal extraction supernatant (LLW) prepared according to Comparative Example 1 of the present invention, Figure 1b is a molecular weight graph for the hydrothermal extract fraction (LLW-S) prepared according to Comparative Example 2 of the present invention 1C is a molecular weight graph of the lotus leaf-derived polysaccharide fraction (LLW-P1) using ethanol precipitation prepared according to Example 1 of the present invention, and FIG. 1D illustrates the ultrafiltration prepared according to Example 2 of the present invention. Molecular weight graph for the lotus leaf-derived polysaccharide fraction (LLW-P2).
Figure 2 is a lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, lotus leaf-derived polysaccharide fraction (LLW-P2), using ultrafiltration prepared according to Example 2, comparison It is a graph measuring the cell viability of the hydrothermal extraction supernatant (LLW) prepared according to Example 1 and the hydrothermal extraction fraction (LLW-S) prepared according to Comparative Example 2.
Figure 3 is a lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, lotus leaf-derived polysaccharide fraction (LLW-P2), using ultrafiltration prepared according to Example 2, comparison Nitrogen oxide (NO) generation capacity for the hydrothermal extraction supernatant (LLW) prepared according to Example 1 and the hydrothermal extraction fraction (LLW-S) prepared according to Comparative Example 2 is measured.
4 is a lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, a lotus leaf-derived polysaccharide fraction (LLW-P2) using ultrafiltration prepared according to Example 2, comparison It is a graph measuring the IL-6 level for the hydrothermal extraction supernatant (LLW) prepared according to Example 1 and the hydrothermal extraction fraction (LLW-S) prepared according to Comparative Example 2.
5 is a lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, a lotus leaf-derived polysaccharide fraction (LLW-P2) using ultrafiltration prepared according to Example 2, comparison It is a graph measuring the TNF-α production capacity for the hydrothermal extraction supernatant (LLW) prepared according to Example 1 and the hydrothermal extraction fraction (LLW-S) prepared according to Comparative Example 2.

The present invention relates to a lotus leaf-derived polysaccharide fraction and a method for preparing the same, which can be ingested in the form of food because it has excellent immune function and no toxicity, thereby improving immune function through diet and lifestyle in daily life. .

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The lotus leaf used as a raw material of the lotus leaf-derived polysaccharide fraction having an immune function-promoting activity of the present invention is collected from early June to mid-October as possible. Usually, the lotus leaf starts to appear in early June and fades when the mid-October leaves. Therefore, it is collected before use as much as possible, and its size is similar to the shape of a half strip of about 40 cm in diameter. The lotus leaf, called the lower lobe, is mainly used in a dried form. It has a bitter taste and has good properties, and has been used since ancient times. It has been used as a folk remedy.

Lotus leaf-derived polysaccharide fraction of the present invention comprises the steps of (A) extracting a mixture of lotus leaf powder and a solvent to obtain an extract; And (B) fractionating the extract by a solvent immersion or non-heat treatment process.

First, in step (A), the lotus leaf powder and the solvent are mixed at a weight ratio of 1:20 to 50, and then extracted at 90 to 110 ° C for 2 to 8 hours, preferably 3 to 5 hours, and then the extract is 3 to 7 ° C. Centrifuge at 5000 to 8000xg for 10 to 30 minutes, then remove the residue and separate the supernatant.

The solvent may be water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

Next, in the step (B), the extract (or the separated supernatant) is precipitated in an organic solvent such as ethanol and methanol and fractionated or fractionated by a non-heat treatment process.

For example, the method of precipitating and fractionating the lotus leaf supernatant in an organic solvent, such as ethanol and methanol, adds an organic solvent to the supernatant so that the final concentration is 70 to 75% and then 10 to 30 hours at -10 to -30 ℃. Precipitating the polysaccharide while standing for a while and then recovering the polysaccharide precipitate by centrifugation for 10 to 30 minutes at 5000 to 8000xg at 3 to 7 ℃ and dialysis it.

As the organic solvent, 70 to 95% of an organic solvent aqueous solution is preferable.

In another example, the lotus leaf supernatant is treated with a non-thermal treatment step, preferably an ultrafiltration device.

The lotus leaf-derived polysaccharide fraction is a polysaccharide having a molecular weight of 5 to 550 KDa at least 50% by weight, preferably at least 60% by weight, more preferably at least 70% by weight, most preferably at least 80% by weight, In one example, the lotus leaf-derived polysaccharide fraction containing this content of polysaccharide is obtained from an ultrafiltration apparatus equipped with a 30 KDa filtration membrane, and is preferably a large molecular weight fraction that does not pass through the 30 KDa filtration membrane. The polysaccharide means a combination of a medium polysaccharide and an acid polysaccharide.

The lotus leaf-derived polysaccharide fraction is 45 to 70% by weight of the neutral polysaccharide (neutral sugar), 20 to 45% by weight of acidic polysaccharides (uronic acid) and 0.5 to 10% by weight of KDO-like substances, 3 to 20% by weight of protein and 0.1 to 5 weight percent phenol.

The acidic polysaccharide consists of galacturonic acid and glucuronic acid; The neutral polysaccharide is composed of arabose (arabinose), rhamnose (rhamnose), galactose (galactose), xylose, glucose (glucose), mannose (fucose) (fucose).

In addition, the KDO-like substance is composed of 3-deoxy-D-lyxo-2-heptulosaric acid (DHA) and 3-deoxy-D-manno-2-octulosonic acid (KDO).

The composition containing the lotus leaf-derived polysaccharide fraction as an active ingredient may be a pharmaceutical composition for the prevention or treatment of diseases caused by a decrease in immunity, a health food composition for enhancing immunity or a cosmetic composition for improving immune function.

As used herein, the term 'fraction' used while referring to lotus leaf includes not only the fraction obtained by treating the extractant but also the processed product of the lotus leaf-derived polysaccharide fraction. For example, the lotus leaf-derived polysaccharide fraction can be prepared in powder form by additional processes such as distillation under reduced pressure and freeze drying or spray drying.

In addition, the lotus leaf-derived polysaccharide fraction of the present invention is broadly meant to include a processed product of lotus leaf, such as lotus leaf powder, which is formulated to administer the lotus leaf to an animal. Although the experiment was conducted with the lotus leaf-derived polysaccharide fraction in the present invention, it will be expected to those skilled in the art that the desired effect can be achieved in the same form as the processed material of the lotus leaf.

On the other hand, the term 'containing as an active ingredient' herein means containing an amount sufficient to achieve the efficacy or activity of the lotus leaf-derived polysaccharide fraction. In one example, the lotus leaf-derived polysaccharide fraction is used at a concentration of 10 to 1500 μg / ml, preferably 100 to 1000 μg / ml. Since the lotus leaf-derived polysaccharide fraction has no adverse effects on the human body even when excessively administered as a natural product, the upper limit of the quantity of the lotus leaf-derived polysaccharide fraction included in the composition of the present invention may be selected and performed by those skilled in the art within an appropriate range.

The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant may include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, lubricants. Agents, flavors and the like can be used.

The pharmaceutical composition may be preferably formulated into a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers in addition to the active ingredient described above for administration.

Formulation forms of the pharmaceutical composition may be granules, powders, tablets, external preparations, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation in the form of tablets or capsules (oral), the active ingredient may be combined with an oral, nontoxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, corn sweeteners, acacia, trackercance or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

Acceptable pharmaceutical carriers in compositions formulated in liquid solutions are sterile and physiologically compatible, including saline, sterile water, Ringer's solution, buffered saline, albumin injectable solutions, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

Furthermore, the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA can be formulated according to each disease or component according to the appropriate method in the art.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, or the like, and preferably, oral administration.

Suitable dosages of the pharmaceutical compositions of the present invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to reaction, and usually The skilled practitioner can readily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dose of the pharmaceutical composition of the present invention is 0.001-10 g / kg.

The pharmaceutical compositions of the present invention may be prepared in unit dose form or formulated into pharmaceutically acceptable carriers and / or excipients or incorporated into multi-dose containers. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or an aqueous medium, or may be in the form of extracts, powders, granules, tablets or capsules, and may further include a dispersant or stabilizer.

In addition, the present invention provides a food composition for improving immune function containing the lotus leaf-derived polysaccharide fraction as an active ingredient.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition, used as a functional food, or added to various foods. Examples of the food to which the composition of the present invention may be added include beverages, alcoholic beverages, confectionary, diet bars, dairy products, meat, chocolates, pizza, ramen noodles, other noodles, gums, ice creams, vitamin complexes, and health supplements. Etc.

The food composition of the present invention may include not only the lotus leaf-derived polysaccharide fraction as an active ingredient, but also components commonly added in food preparation, and include, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. do. Examples of the above carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And sugars such as conventional sugars such as polysaccharides such as dextrin, cyclodextrin and the like and xylitol, sorbitol, erythritol. As the flavoring agent, natural flavoring agents (tauumatin, stevia extract (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared with a drink and a beverage, in addition to the lotus leaf-derived polysaccharide fraction of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be further included. have.

The present invention provides a health functional food containing a food composition comprising the lotus leaf-derived polysaccharide fraction as an active ingredient. Health functional foods are foods prepared by adding lotus leaf-derived polysaccharide fractions to food materials such as beverages, teas, spices, gums, confectionery, or by encapsulating, powdering, and suspensions. It means coming, but unlike the general medicine has the advantage that there are no side effects that can occur when taking a long-term use of the drug as a raw material. The health functional food of the present invention thus obtained is very useful because it can be consumed on a daily basis. The amount of the lotus leaf-derived polysaccharide fraction in such a dietary supplement can not be uniformly defined depending on the type of dietary supplement, but may be added within a range that does not impair the original taste of the dietary supplement. It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of a health functional food in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the dietary supplement of the present invention may be in the form of pills, tablets, capsules or beverages.

The present invention also provides the use of a lotus leaf-derived polysaccharide fraction for the manufacture of a medicament or food for improving human immune function. As described above, the lotus leaf-derived polysaccharide fraction may be used for the purpose of improving immune function.

The present invention also provides a method for improving immune function comprising administering to a mammal an effective amount of a lotus leaf-derived polysaccharide fraction.

As used herein, the term "mammal" refers to a mammal that is the subject of treatment, observation or experimentation, preferably human.

As used herein, the term “effective amount” means an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, doctor or other clinician, Amounts that induce alleviation of the symptoms of the disease or disorder. The effective amount and frequency of administration for the active ingredient of the present invention may vary depending on the desired effect. Therefore, the optimal dosage to be administered can be readily determined by one skilled in the art and includes the type of disease, the severity of the disease, the amount of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, general health of the patient. It may be adjusted according to various factors including the condition, sex and diet, time of administration, route of administration and the rate of secretion of the composition, the duration of treatment, and the drugs used simultaneously. In the prophylactic, treatment or improvement method of the present invention, in the case of an adult, when the extract of leek leaf, leaf leaf or mixture of leeks is administered once to several times a day, at a dose of 0.001 g / kg to 10 g / kg It is preferable to administer.

The composition comprising the lotus leaf-derived polysaccharide fraction as an active ingredient in the treatment method of the present invention is conventional through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. May be administered in such a manner.

In addition, the cosmetic composition of the present invention may be blended with the cosmetic composition described above, if necessary, with other ingredients normally blended into the cosmetic composition. Examples of such ingredients include water, oil, surfactants, moisturizers, thickeners, fragrances, preservatives, And at least one member selected from the group consisting of neutralizers, emollients, skin protectants, and skin conditioning agents.

In addition, the cosmetic composition may be prepared in the form of an emulsified formulation and a solubilized formulation commonly used in the art, for example, skin, latex, essence, lotion, beauty liquid, body lotion, body gel, body essence, body cleanser, cleansing Foam, cleansing cream, cleansing gel, pack, massage cream, massage gel, nutrition cream, sleeping cream and moisture cream any one selected from the group consisting of.

Hereinafter, preferred examples are provided to aid the understanding of the present invention, but the following examples are merely for exemplifying the present invention, and it will be apparent to those skilled in the art that various changes and modifications can be made within the scope and spirit of the present invention. It is natural that such variations and modifications fall within the scope of the appended claims.

Example 1. Lotus Leaf-derived Polysaccharide Fraction (LLW-P1) Using Ethanol Precipitation

The lotus leaf was purchased from Gyeongdong Market (Korea) in Seoul and used as a sample after coarse grinding. 30 times (w / v) of distilled water was added to the coarsely pulverized lotus leaf, and heated at 90 ° C. for 4 hours. The hot water extract was centrifuged at 4 ° C. and 6,000 kg for 20 minutes, and then the residue was removed. (LLW) was isolated.

Three times the volume (v / v) of alcohol was added to the separated supernatant so that the final concentration was 75%, and the polysaccharide was allowed to stand at -20 ° C. overnight (12 hours). At this time, the precipitate was centrifuged at 6,000Xg for 20 minutes, and then the material except for the supernatant (Comparative Example 2) was recovered, redissolved in a small amount of distilled water, and dialyzed for 3 days (MWCO: 6,000-8,000 Da). Lyophil-derived crude polysaccharide fraction (LLW-P1) was obtained by lyophilization (Tokoyo Rikakikai Co. Ltd, FD-1000, Tokyo, Japan).

Example 2. Lotus Leaf-derived Polysaccharide Fractions Using Ultrafiltration (LLW-P2)

The lotus leaf was purchased from Gyeongdong Market (Korea) in Seoul and used as a sample after coarse grinding. 30 times (w / v) of distilled water was added to the coarsely pulverized lotus leaf, and heated at 90 ° C. for 4 hours. The hot water extract was centrifuged at 4 ° C. and 6,000 × g for 20 minutes, and the residue was removed. LLW) was isolated.

The separated supernatant was freeze-dried by separating a material that did not pass through an ultrafiltration apparatus (Sartocon-Mini system, Sartorius Co., G ㆆ ttingen, Germany) using a filtration membrane having a molecular weight cut off (MWCO; 30 KDa). (Tokoyo Rikakikai Co. Ltd, FD-1000, Tokyo, Japan) to obtain a lotus leaf derived crude polysaccharide fraction (LLW-P2).

The ultrafiltration device is a laboratory scale ultrafiltration device (Sartocon-Mini system, Sartorius Co., G ㆆ ttingen, consisting of a 1 L feed tank, a feed pump, a filtration cartridge and a pressure gauge for the feed and the ultrafiltration permeate). Germany).

Comparative Example 1. Hot Water Extracting Supernatant (LLW)

The supernatant (LLW) prepared in Example 1 was used.

Comparative Example 2. Hot Water Extract Fraction (LLW-S)

Ethanol was added in Example 1, the mixture was allowed to stand at -20 ° C. overnight (12 hours), and the supernatant separated by centrifugation from the precipitate was filtered and lyophilized to obtain a hot water extract fraction (LLW-S).

<Test Example>

Test Example 1. Molecular Weight Measurement

Figure 1a is a molecular weight graph for the hydrothermal extraction supernatant (LLW) prepared according to Comparative Example 1 of the present invention, Figure 1b is a molecular weight graph for the hydrothermal extract fraction (LLW-S) prepared according to Comparative Example 2 of the present invention 1C is a molecular weight graph of the lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, and FIG. 1D is an ultrafiltration prepared according to Example 2 of the present invention. Molecular weight graph for the lotus leaf-derived polysaccharide fraction (LLW-P2) using.

0.45 μm filter paper (Toyo Roshi Kaisha, Ltd., Tokyo, Japan) diluted with distilled water at a concentration of 20 mg / mL to determine the purity and molecular weight of the lotus leaf-derived crude polysaccharide fractions prepared according to the examples and comparative examples. After filtration through an Asahi-Pak GS-520 + GS-320 packed column (0.76 X 30 cm, Showa denco Co., Tokyo, Japan, respectively) equilibrated with 50 mM ammonium formate (pH 5.5) Analysis was performed using a high performance liquid chromatography (HPLC) (Agilent Technologies Co., Santa Clara, CA, USA). Molecular weight was determined by using the standard pullulan series (1.32, 6, 10, 48.8, 113, 366K) to determine the retention time, and then converted from the standard curve created by calculating the Kav value for each molecular weight.

As shown in Figures 1a to 1d, the lotus leaf-derived polysaccharide fraction (LLW-P1, yield: 0.64) using the ethanol precipitation method of Example 1 is a relatively purified lotus leaf-derived polymer material is removed low molecular weight of less than 1 kDa It was confirmed that it contained 20-50 kDa and 5-15 kDa as the main peak. In addition, the lotus leaf-derived polysaccharide fraction using ultrafiltration of Example 2 (LLW-P2, yield: 0.122%) was able to more effectively remove the low molecular weight of less than 1 kDa, mainly containing a polymer of 5-15 kDa or more It was confirmed.

On the other hand, the hydrothermal extraction supernatant of Comparative Example 1 (LLW, yield: 12.01%) was confirmed to be a low molecular weight of less than 1 kDa, the hydrothermal extraction fraction of Comparative Example 2 (LLW-S, yield 10.81%) also low molecular weight of less than 1 kDa As the main component of water, the low molecular weight contained in the LLW was found to migrate to the LLW-S fraction.

Test Example 2 Component Analysis

Neutral polysaccharide content is galactose (galactose) as a reference material phenol-sulfuric acid method (Dubois et al., 1956), acidic polysaccharide content (galacturonic acid) as a standard m- M-hydroxybiphenyl method (Blumenkrantz & Asboe-Hansen, 1973), protein content using bovine serum albumin as standard, Bradford method (Bradford MM, 1976), TBA- The positive material content was 2-keto-3-deoxy-D-manno-octulosonic acid (KDO) as a reference material, and thiobarbituric acid colorimetric method (Karkhanis et al., 1978) was used to modify the laboratory conditions. In addition, the total phenol content is based on the fact that the folin-Ciocalteu reagent is reduced by the polyphenolic compound of the extract to form molybdenum blue according to the method of Singleton et al. (1999) using gallic acid as a standard. Was measured.

[Table 1] shows the content of each component, and [Table 2] shows each content in weight percent.

Division (unit: mg / g) Example 1 Example 2 Comparative Example 1 Comparative Example 2 Neutral polysaccharide 401.2 ± 1.71 483.2 ± 7.22 327.8 ± 4.05 321.0 ± 2.97 Acid polysaccharide 208.3 ± 2.88 296.8 ± 3.73 123.8 ± 0.61 119.3 ± 7.22 protein 42.0 ± 1.39 66.1 ± 1.89 23.4 ± 2.65 25.2 ± 0.95 KDO analogue 6.64 ± 0.17 12.6 ± 0.50 5.02 ± 0.09 4.70 ± 0.21 Total phenol 2.38 ± 0.08 6.83 ± 0.79 6.88 ± 0.27 7.43 ± 0.16 Total content 660.5 ± 5.76 865.5 ± 3.74 486.8 ± 3.80 477.6 ± 9.33

Division (unit: weight%) Example 1 Example 2 Comparative Example 1 Comparative Example 2 Neutral polysaccharide 60.7 ± 0.27 55.8 ± 0.62 67.3 ± 0.65 67.2 ± 0.70 Acid polysaccharide 31.5 ± 0.16 34.3 ± 0.50 25.4 ± 0.22 25.0 ± 1.04 protein 6.36 ± 0.16 7.64 ± 0.25 4.80 ± 0.53 5.27 ± 0.30 KDO analogue 1.00 ± 0.03 1.46 ± 0.06 1.03 ± 0.02 0.98 ± 0.06 Total phenol 0.36 ± 0.01 0.79 ± 0.09 1.41 ± 0.06 1.55 ± 0.02 Total content 100 100 100 100

 As shown in Tables 1 and 2 above, the lotus leaf-derived copolysaccharide fractions prepared according to Examples 1 and 2 have a higher content of neutral polysaccharides, acidic polysaccharides, proteins and KDO-like substances than the fractions of Comparative Examples 1 and 2. In addition, it was confirmed that the total component content was also high.

Calculating the weight percentage of each substance, the content ratio of acidic polysaccharides contained in the lotus leaf-derived copolysaccharide fractions prepared according to Examples 1 and 2 was higher than that of the acidic polysaccharides contained in the fractions of Comparative Examples 1 and 2. It was confirmed.

Test Example 3 RAW 264.7 Cell Proliferation Effect

Cell culture

RAW 264.7 cells, a macrophage cell line of mice, were used by the Korea Cell Line Bank (KCLB). The RAW 264.7 cells were incubated at 37 ° C., 5% CO ₂ using Dulbecco 'modified Eagle' medium (DMEM: Gibco BRL, GrandIsland, NY, USA) medium containing 10% FBS and 1% penicillin-streptomycin. Incubated at.

3.1 Measuring Cell Viability

Figure 2 is a lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, lotus leaf-derived polysaccharide fraction (LLW-P2), using ultrafiltration prepared according to Example 2, comparison It is a graph measuring the cell viability of the hydrothermal extraction supernatant (LLW) prepared according to Example 1 and the hydrothermal extraction fraction (LLW-S) prepared according to Comparative Example 2.

To measure the cytotoxicity level of the RAW 264.7 cells, the cell viability was measured using CCK-8 assay. RAW 264.7 cells were dispensed in 96-well plates at a concentration of 2 × 10 ⁵ cells / mL and incubated for 24 hours. Each sample was treated at a concentration of 1, 10, 100 μg / mL. In addition, the positive control group was treated with 1 μg / mL lipopolysaccharide (LPS), and the non-treated control group was treated with DMEM medium for 20 hours and further cultured with Ez-Cytox cell viability assay kit (Daeillab service co., LTD. , Korea) was used for 90 minutes at 37 ℃ to measure the absorbance at 450 nm.

In order to confirm the cytotoxicity of the sample was treated by the concentration of 1, 10, 100 μg / mL as shown in Figure 2, as shown in Figure 2, derived from the lotus leaf of Example 1 (LLW-P1) and Example 2 (LLW-P2) When the crude polysaccharide fraction was treated at a concentration of 1-100 μg / mL, all concentrations showed significantly higher cell viability compared to the normal control group (NC).

On the other hand, Comparative Example 1 (LLW) and Comparative Example 2 (LLW-S) showed a survival rate of less than 100% at low concentration, but it was confirmed that there is no significant difference from the normal control group.

3.2 Measurement of nitric oxide (NO) formation ability

Figure 3 is a lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, lotus leaf-derived polysaccharide fraction (LLW-P2), using ultrafiltration prepared according to Example 2, comparison Nitrogen oxide (NO) generation capacity for the hydrothermal extraction supernatant (LLW) prepared according to Example 1 and the hydrothermal extraction fraction (LLW-S) prepared according to Comparative Example 2 is measured.

When macrophages are activated, they secrete cytokines such as nitric oxide (II) and TNF-α, where the production of nitric oxide (II) is an important signaling material that acts as a defense against tumor cells or intracellular infected microorganisms in the immune system. Known as In this case, the excessive production of nitric oxide (II) causes systemic inflammation and has various negative effects on the living body, but the production of an appropriate amount of nitric oxide (II) is considered as an important factor of innate immunity.

Griess reagent [1% (w / v) sulfanilamide in 5% (v / v) phosphoric acid and 0.1% (w / v) naphtylethylenediamine-HCl )] Was used to measure the concentration of NO ₂ ⁻ present in the cell culture. That is, RAW 264.7 cells were dispensed in 48-well plates at a concentration of ⁵ × 10 ⁵ cells / well and incubated for 24 hours, and then the samples were treated by concentrations (0, 10, 100 μg / mL). At this time, as a positive control lipopolysaccharide (LPS) was 1 μg / mL, and as an untreated control group was further cultured for 20 hours by treatment with DMEM medium. The supernatant of the cultured cell culture solution and the same amount of griess reagent were mixed and reacted at room temperature for 15 minutes, and the absorbance was measured at 540 nm. The concentration of nitric oxide (II) was determined from a standard curve prepared using sodium nitrite. Obtained and converted.

Samples Example 1 (LLW-P1), Example 2 (LLW-P2), Comparative Example 1 (LLW) and Comparison with the positive control group treated only with normal control group (NC) and lipopolysaccharide (LPS, 1 μg / mL) Example 2 (LLW-S) by the concentration (1, 10, 100 μg / mL) when the amount of nitric oxide (II) produced by measuring the results are shown in FIG.

As shown in FIG. 3, the lotus leaf-derived copolysaccharide fraction of Example 1 (LLW-P1) was significantly increased at a concentration of 10 μg / mL or more, especially at a concentration of 26.8 μM at 100 μg / mL. Confirmed. In addition, the lotus leaf-derived copolysaccharide fraction of Example 2 (LLW-P2) showed a significant increase by concentration gradient at a concentration of 1 μg / mL or more and was confirmed to be very high at a level of 37.5 μM at a concentration of 100 μg / mL.

On the other hand, in Comparative Example 1 (LLW) and Comparative Example 2 (LLW-S) compared to the normal control group (6.3 μM) in all treatment concentrations of 1-100 μg / mL significantly compared to the normal control (NC) There was no difference.

3.3 Measurement of Promoting Cytokine Production

4 is a lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, a lotus leaf-derived polysaccharide fraction (LLW-P2) using ultrafiltration prepared according to Example 2, comparison It is a graph measuring the IL-6 level for the hydrothermal extraction supernatant (LLW) prepared according to Example 1 and the hydrothermal extraction fraction (LLW-S) prepared according to Comparative Example 2.

In addition, Figure 5 is a lotus leaf-derived polysaccharide fraction (LLW-P1) using the ethanol precipitation method prepared according to Example 1 of the present invention, a lotus leaf-derived polysaccharide fraction using ultrafiltration prepared according to Example 2 (LLW-P2) , TNF-α generating ability for the hydrothermal extraction supernatant (LLW) prepared according to Comparative Example 1 and the hydrothermal extraction fraction (LLW-S) prepared according to Comparative Example 2 is measured.

Activated macrophages are known to induce subsequent immune responses by producing cytokines such as TNF-α, interleukin-6, and interleukin-12, which are considered important factors of the innate immune response. It acts as a stimulating factor for B cells, which induces the final differentiation of plasma cells, and promotes the synthesis of immunoglobulins.

To determine the degree of activation of the immune response by measuring the amount of IL-6 and TNF-α secreted when macrophages treated with the lotus leaf-derived polysaccharide fraction of the present invention by concentration (1, 10, 100 μg / mL).

The measurement of the production of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the cell culture was carried out by 24 hours after dispensing RAW 264.7 cells in a 48-well plate at a concentration of 2 × 10 ⁵ cells / mL. Was treated at a concentration of 1, 10, 100 μg / mL, incubated for 20 hours, and the absorbance of IL-6 and TNF-α secreted into the supernatant was measured at 450 nm using an ELISA kit (BD Biosciences, USA). It was.

Samples Example 1 (LLW-P1), Example 2 (LLW-P2), Comparative Example 1 (LLW) and Comparison with the positive control group treated only with normal control group (NC) and lipopolysaccharide (LPS, 1 μg / mL) As a result of measuring the IL-6 level generated when Example 2 (LLW-S) was treated by concentration (1, 10, 100 μg / mL) is shown in FIG.

As shown in Figure 4, the lotus leaf-derived copolysaccharide fraction of Example 1 (LLW-P1) significantly increased IL-6 levels at concentrations of 10 μg / mL or higher, particularly at 5161.1 pg at 100 μg / mL. It was confirmed that the / mL level was very high. In addition, the lotus leaf-derived crude polysaccharide fraction of Example 2 (LLW-P2) showed a concentration-dependent IL-6 increase pattern in the treatment concentration range of 1-100 μg / mL, especially 5306.4 pg / mL at 100 μg / mL. Levels were found to be significantly similar to the positive control group treated with LPS (5345.7 pg / mL).

On the other hand, Comparative Example 1 (LLW) showed no significant difference in IL-6 production ability compared to the normal control group (17.0 μg / mL), Comparative Example 2 (LLW-S) 64.9 pg at 100 μg / mL concentration / mL level showed a slight increase in IL-6 production.

In addition, the results of examining the immune activity of the inflammatory cytokine TNF-α production ability is shown in FIG.

As shown in FIG. 5, the lotus leaf-derived copolysaccharide fraction of Example 1 (LLW-P1) was 1662.0 μg / mL, 10290.0 μg / mL, and 17924.5 μg /, respectively at concentration ranges of 1, 10, and 100 μg / mL. It was confirmed that the TNF-α producing ability was high at the mL level. In particular, the TNF-α production level at the concentration of 100 μg / mL of Example 1 (LLW-P1) was significantly similar to the positive control group treated with LPS (17754.3 pg / mL).

In addition, the lotus leaf-derived crude polysaccharide fraction of Example 2 (LLW-P2) also showed a TNF-α increase pattern in a concentration-dependent manner in the range of 1-100 μg / mL, the level is 15066.0 μg / mL, 15905.7 μg / mL, respectively , And 16305.7 μg / mL showed very high TNF-α production ability.

On the other hand, Comparative Example 1 (LLW) compared to the normal control group (227.3 μg / mL) was significantly higher TNF-α to 968.0 μg / mL and 5886.3 μg / mL at 10 μg / mL and 100 μg / mL concentration, respectively Indicated levels. In addition, Comparative Example 2 (LLW-S) did not show a TNF-α increase level in all concentration range of 1-100 μg / mL.

Hereinafter, an example of the preparation of the composition containing the powder of the present invention will be described, but the present invention is not intended to limit the present invention, but is only intended to be described in detail.

Formulation Example 1 Preparation of Powder

500 mg of crude polysaccharide fraction derived from lotus leaf obtained in Example 1

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

300 mg of crude polysaccharide fraction derived from lotus leaf obtained in Example 1

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

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

Formulation Example 3 Preparation of Capsule

200 mg of crude polysaccharide fraction derived from lotus leaf obtained in Example 1

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

600 mg of crude polysaccharide fraction derived from lotus leaf obtained in Example 1

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO _4, 12H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule.

Formulation Example 5 Preparation of Liquid

4 g of crude polysaccharide fraction derived from lotus leaf obtained in Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water

After dissolving each component in purified water according to the usual method of preparing a liquid solution, adding a lemon flavor, and then mixing the above ingredients, adding purified water to adjust the total to 100g by adding purified water, and then filling into a brown bottle to sterilize To prepare a liquid solution.

Formulation Example 6 Preparation of Granules

1,000 mg of crude polysaccharide fraction derived from lotus leaf obtained in Example 1

Vitamin Mixture

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

0.2 μg of vitamin B12

Vitamin C 10 mg

10 μg biotin

Nicotinamide 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5 mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

15 mg potassium monophosphate

Dicalcium Phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is a composition which is relatively suitable for granules in a preferred embodiment, the composition ratio may be arbitrarily modified, and the above components are mixed according to a conventional granulation method and then granulated. It can be prepared and used in the manufacture of health functional food composition according to a conventional method.

Formulation Example 7 Preparation of Functional Beverage

 1,000 mg of crude polysaccharide fraction derived from lotus leaf obtained in Example 1

Citric Acid 1,000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 mL of purified water

After mixing the above components in accordance with the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and refrigerated Used to prepare functional beverage compositions of the invention.

Although the composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

Formulation Example 8 Preparation of Cosmetics in Emulsion Formulation

Cosmetics of emulsion formulations such as nutrient cosmetics, creams and essences, and cosmetics of solubilized formulations such as softening cosmetics were prepared.

To the cosmetic composition was prepared in the composition shown in Table 3. The manufacturing method is as follows.

1) The mixture which mixed the raw materials of 1-9 was heated at 65-70 degreeC.

2) 10 was added to the mixture of step 1).

3) The mixture of the raw materials of 11-13 was heated to 65-70 degreeC, and completely dissolved.

4) Through step 3), the mixture of 2) was slowly added to emulsify for 2 to 3 minutes at 8,000 rpm.

5) The raw material of 14 was dissolved in a small amount of water, and then added to the mixture of step 4) and emulsified for 2 minutes.

6) 15 to 17 of the raw materials were weighed, respectively, and added to the mixture of step 5) and emulsified for 30 seconds.

7) After the emulsification of the mixture of step 6) through a degassing process to cool to 25 ~ 35 ℃ to prepare an emulsion cosmetic.

Furtherance Emulsifier 1 Emulsifier 2 Emulsifier 3 One Stearic acid 0.3 0.3 0.3 2 Steali alcohol 0.2 0.2 0.2 3 Glyceryl Monostearate 1.2 1.2 1.2 4 Beeswax 0.4 0.4 0.4 5 Polyoxyethylene sorbitan
Monolauric acid ester 2.2 2.2 2.2 6 Methyl paraoxybenzoate 0.1 0.1 0.1 7 Paraoxybenzoic Acid Profiles 0.05 0.05 0.05 8 Cetylethylhexanoate 5 5 5 9 Triglycerides 2 2 2 10 Cyclomethicone 3 3 3 11 Distilled water To 100 To 100 To 100 12 Concentrated glycerin 5 5 5 13 Triethanolamine 0.15 0.15 0.15 14 Polyacrylic acid polymer 0.12 0.12 0.12 15 Pigment 0.0001 0.0001 0.0001 16 incense 0.10 0.10 0.10 17 Fractions of Example 1 0.0001 One 10

Formulation Example 9 Preparation of Cosmetics in Solubilized Formulations

To the cosmetic composition of the solubilizing agent type was prepared in the composition shown in Table 4. The manufacturing method is as follows.

1) Raw materials 2 to 6 were placed in raw material 1 (purified water) and dissolved using a mixer.

2) The raw materials of 8-11 were put into the raw material of 7 (alcohol), and it melt | dissolved completely.

3) The mixture of step 2) was solubilized with slow addition to the mixture of step 1).

Furtherance Solubilized Formulation 1 Solubilized Formulation 2 Solubilized Formulation 3 One Purified water To 100 To 100 To 100 2 Concentrated glycerin 3 3 3 3 1,3-butylene glycol 2 2 2 4 EDTA-2Na 0.01 0.01 0.01 5 Pigment 0.0001 0.0002 0.0002 6 Fractions of Example 1 0.1 5 5 7 Alcohol (95%) 8 8 8 8 Methyl paraoxybenzoate 0.1 0.1 0.1 9 Polyoxyethylene
Hydrogenated Ester 0.3 0.3 0.3 10 incense 0.15 0.15 0.15 11 Cyclomethicone - - 2

Claims (14)

45 to 70% by weight neutral sugar, 20 to 45% by weight uronic acid, 0.5 to 10% by weight KDO analogue, 3 to 20% by weight protein and 0.1 to 5% by weight phenol relative to the total polysaccharide fraction Lotus leaf-derived polysaccharide fraction comprising a. The method of claim 1, wherein the neutral polysaccharide comprises arabinose, rhamnose, galactose, xylose, glucose, mannose and fucose. Lotus leaf-derived polysaccharide fraction, characterized in that. According to claim 1, wherein the acidic polysaccharide is lotus leaf-derived polysaccharide fraction, characterized in that consisting of galacturonic acid (gluacturonic acid) and glucuronic acid (glucuronic acid). According to claim 1, wherein the KDO-like substance lotus leaf characterized in that it comprises 3-deoxy-D-lyxo-2-heptulosaric acid (DHA) and 3-deoxy-D-manno-2-octulosonic acid (KDO) Derived polysaccharide fraction. According to claim 1, wherein the lotus leaf-derived polysaccharide fraction is lotus leaf-derived polysaccharide fraction, characterized in that the molecular weight of 5 to 550 KDa polysaccharide containing 50% by weight or more. According to claim 1, wherein the lotus leaf-derived polysaccharide fraction is lotus leaf-derived polysaccharide fraction, characterized in that it has an immune function enhancing activity. (A) mixing and extracting the lotus leaf powder and a solvent to obtain an extract; And
(B) fractionating the extract by a solvent immersion or non-heat treatment process; a method for producing a lotus leaf-derived polysaccharide fraction comprising a. The method according to claim 7, wherein the lotus leaf-derived polysaccharide fraction fractionated in step (B) is 45 to 70% by weight neutral sugar, 20 to 45% by weight acidic polysaccharide (uronic acid), KDO The analogous material comprises 0.5 to 10% by weight, 3 to 20% by weight protein and 0.1 to 5% by weight phenol. The method according to claim 7, wherein the lotus leaf-derived polysaccharide fraction fractionated in step (B) is a method for producing a lotus leaf-derived polysaccharide fraction, characterized in that the polysaccharide has a molecular weight of 5 to 550 KDa. 8. The method of claim 7, wherein the solvent in step (A) is water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The pharmaceutical composition for the prevention or treatment of diseases caused by a decrease in immunity, comprising the lotus leaf-derived polysaccharide fraction of claim 1 as an active ingredient. The pharmaceutical composition for preventing or treating a disease caused by a lowered immune system according to claim 11, wherein the disease caused by a lowered immune function is selected from the group consisting of an infectious disease, an inflammatory disease, an allergic disease, or chronic fatigue. Health food composition for enhancing immune function, comprising the lotus leaf-derived polysaccharide fraction of claim 1 as an active ingredient. Claim 1, a cosmetic composition for enhancing immune function, comprising the lotus leaf-derived polysaccharide fraction as an active ingredient.

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